From 6bbdfbce86d1aa086c79a738bdda336b723ca438 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 18 May 2020 08:41:35 +0200 Subject: [PATCH 01/99] Ignoring Aux folder --- .Rbuildignore | 3 ++- .gitignore | 3 ++- 2 files changed, 4 insertions(+), 2 deletions(-) diff --git a/.Rbuildignore b/.Rbuildignore index e9333c4..25ff9f8 100644 --- a/.Rbuildignore +++ b/.Rbuildignore @@ -15,4 +15,5 @@ postBuild runtime.txt .travis.yml .github -.git \ No newline at end of file +.git +Aux \ No newline at end of file diff --git a/.gitignore b/.gitignore index 666540e..50a8bc7 100644 --- a/.gitignore +++ b/.gitignore @@ -1,3 +1,4 @@ inst/doc doc -Meta \ No newline at end of file +Meta +Aux \ No newline at end of file From a01da43bb52a025e90c58ce8603158d9d015f34d Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 18 May 2020 09:57:17 +0200 Subject: [PATCH 02/99] Added minimal (freezing) test --- tests/testthat/test.DEGfreeze.R | 49 +++++++++++++++++++++++++++++++++ 1 file changed, 49 insertions(+) create mode 100644 tests/testthat/test.DEGfreeze.R diff --git a/tests/testthat/test.DEGfreeze.R b/tests/testthat/test.DEGfreeze.R new file mode 100644 index 0000000..c0c1ad3 --- /dev/null +++ b/tests/testthat/test.DEGfreeze.R @@ -0,0 +1,49 @@ +library(devtools)#TEMP +context("Stress-testing DEG analyses") +# ============================================================================== +# Loading dataset +# ============================================================================== +install()#TEMP +library(DIscBIO)#TEMP +data(pan_indrop_matrix_8000cells_18556genes) + +# ============================================================================== +# Determining contants +# ============================================================================== +n_genes <- 500 +n_clust <- 2 + +# ============================================================================== +# Defining datasets to be used +# ============================================================================== +sc <- DISCBIO(pan_indrop_matrix_8000cells_18556genes[, seq_len(n_genes)]) + +# ============================================================================== +# Performing operations +# ============================================================================== +# MIinExp <- mean(rowMeans(dataset, na.rm=TRUE)) +# MinNumber <- round(length(dataset[1, ]) / 10) # 2b xpressed in > 10% cells. +# sc <- Normalizedata( +# sc, mintotal=1000, minexpr=MIinExp, minnumber=MinNumber, maxexpr=Inf, +# downsample=FALSE, dsn=1, rseed=17000 +# ) +sc <- FinalPreprocessing(sc, GeneFlitering="ExpF", export=FALSE, quiet=TRUE) +sc <- Clustexp(sc, cln=n_clust, quiet=TRUE) + +# ------------------------------------------------------------------------------ +# DEG Analysis +# ------------------------------------------------------------------------------ +cdiff <- DEGanalysis( + sc, Clustering="K-means", K=n_clust, fdr=0.10, name="all_clusters", + export=FALSE, quiet=FALSE, plot=FALSE, nresamp=5, nperms=10 +) +# sc <- Exprmclust(sc, K=K, reduce=TRUE, quiet=TRUE) +# sc <- MB_Order(sc, quiet=TRUE, export=FALSE) +# MBcdiffBinomial <- MBClustDiffGenes( +# sc, K=K, pValue=0.05, fdr=0.15, export=FALSE, quiet=FALSE +# ) +# sc <- Clustexp(sc, cln=K, quiet=TRUE) +# MBcdiff <- DEGanalysis( +# sc, Clustering="MB", K=K, fdr=0.05, name="all_clusters", export = FALSE, +# quiet=TRUE +# ) \ No newline at end of file From c4acc2c2ae6bc9b12fea94332025d6ac9810023c Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 18 May 2020 13:58:34 +0200 Subject: [PATCH 03/99] Added draft of DEGfreeze tests --- tests/testthat/test.DEGfreeze.R | 28 ++++++++++++++++++++-------- 1 file changed, 20 insertions(+), 8 deletions(-) diff --git a/tests/testthat/test.DEGfreeze.R b/tests/testthat/test.DEGfreeze.R index c0c1ad3..96c396e 100644 --- a/tests/testthat/test.DEGfreeze.R +++ b/tests/testthat/test.DEGfreeze.R @@ -1,21 +1,27 @@ -library(devtools)#TEMP +# library(devtools)#TEMP +# library(testthat)#TEMP +# source("Aux/samr.R")#TEMP +# source("Aux/resample.R")#TEMP +# source("Aux/rankcol.R")#TEMP +# source("Aux/samr/R/samr.morefuns.R")#TEMP +# source("Aux/samr/R/samr.R")#TEMP +# library(DIscBIO)#TEMP context("Stress-testing DEG analyses") # ============================================================================== # Loading dataset # ============================================================================== -install()#TEMP -library(DIscBIO)#TEMP data(pan_indrop_matrix_8000cells_18556genes) # ============================================================================== # Determining contants # ============================================================================== -n_genes <- 500 -n_clust <- 2 +n_genes <- 1e3 +n_clust <- 3 # ============================================================================== # Defining datasets to be used # ============================================================================== +message(Sys.time(), " - Subsetting") sc <- DISCBIO(pan_indrop_matrix_8000cells_18556genes[, seq_len(n_genes)]) # ============================================================================== @@ -27,15 +33,18 @@ sc <- DISCBIO(pan_indrop_matrix_8000cells_18556genes[, seq_len(n_genes)]) # sc, mintotal=1000, minexpr=MIinExp, minnumber=MinNumber, maxexpr=Inf, # downsample=FALSE, dsn=1, rseed=17000 # ) +message("Filtering") +sc <- Normalizedata(sc, mintotal=1000, maxexpr=Inf, downsample=FALSE, dsn=1) sc <- FinalPreprocessing(sc, GeneFlitering="ExpF", export=FALSE, quiet=TRUE) -sc <- Clustexp(sc, cln=n_clust, quiet=TRUE) +sc <- Clustexp(sc, clustnr=n_clust, quiet=TRUE, bootnr = 2, B.gap = 2) # ------------------------------------------------------------------------------ # DEG Analysis # ------------------------------------------------------------------------------ +message("Analysing DEG") cdiff <- DEGanalysis( sc, Clustering="K-means", K=n_clust, fdr=0.10, name="all_clusters", - export=FALSE, quiet=FALSE, plot=FALSE, nresamp=5, nperms=10 + export=FALSE, quiet=TRUE, plot=FALSE, nresamp=2, nperms=2 ) # sc <- Exprmclust(sc, K=K, reduce=TRUE, quiet=TRUE) # sc <- MB_Order(sc, quiet=TRUE, export=FALSE) @@ -46,4 +55,7 @@ cdiff <- DEGanalysis( # MBcdiff <- DEGanalysis( # sc, Clustering="MB", K=K, fdr=0.05, name="all_clusters", export = FALSE, # quiet=TRUE -# ) \ No newline at end of file +# ) +test_that("DEG analysis doesn't freeze", { + expect_output(str(cdiff), "List of 2") +}) \ No newline at end of file From 5f653fb11e9325b6334b347a12f694801e53115d Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Tue, 26 May 2020 07:45:13 +0200 Subject: [PATCH 04/99] Added minimally-workign code from samr --- DESCRIPTION | 1 + R/samr-adapted.R | 782 ++++++++++++++++++++++++ man/check.format.Rd | 11 + man/foldchange.seq.twoclass.unpaired.Rd | 11 + man/rankcols.Rd | 17 + man/resa.Rd | 21 + man/sammy.Rd | 32 + man/samr.estimate.depth.Rd | 17 + man/wilcoxon.unpaired.seq.func.Rd | 19 + 9 files changed, 911 insertions(+) create mode 100644 R/samr-adapted.R create mode 100644 man/check.format.Rd create mode 100644 man/foldchange.seq.twoclass.unpaired.Rd create mode 100644 man/rankcols.Rd create mode 100644 man/resa.Rd create mode 100644 man/sammy.Rd create mode 100644 man/samr.estimate.depth.Rd create mode 100644 man/wilcoxon.unpaired.seq.func.Rd diff --git a/DESCRIPTION b/DESCRIPTION index 4baa3ac..03af8d9 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -94,3 +94,4 @@ Collate: 'customConverters.R' 'datasets.R' 'retrieveBiomart.R' + 'samr.R' diff --git a/R/samr-adapted.R b/R/samr-adapted.R new file mode 100644 index 0000000..d5151a5 --- /dev/null +++ b/R/samr-adapted.R @@ -0,0 +1,782 @@ +# This script contains customized versions of functions found in the samr package. This is necessary because samr seems to have been abandoned, so an upstream collaboration doesn't seem possible at the time of writing. + +# ============================================================================== +# Constants +# ============================================================================== +samr.const.twoclass.unpaired.response <- "Two class unpaired" +samr.const.twoclass.paired.response <- "Two class paired" +samr.const.oneclass.response <- "One class" +samr.const.quantitative.response <- "Quantitative" +samr.const.multiclass.response <- "Multiclass" +samr.const.twoclass.unpaired.timecourse.response <- "Two class unpaired timecourse" +samr.const.twoclass.paired.timecourse.response <- "Two class paired timecourse" +samr.const.oneclass.timecourse.response <- "One class timecourse" +samr.const.survival.response <- "Survival" +samr.const.patterndiscovery.response <- "Pattern discovery" + +# ============================================================================== +# Functions +# ============================================================================== + +#' @title Significance analysis of microarrays +#' @description This function is an adaptation of `samr::samr` +sammy <- function (data, resp.type = c("Quantitative", "Two class unpaired", + "Survival", "Multiclass", "One class", "Two class paired", + "Two class unpaired timecourse", "One class timecourse", + "Two class paired timecourse", "Pattern discovery"), assay.type = c("array", + "seq"), s0 = NULL, s0.perc = NULL, nperms = 100, center.arrays = FALSE, + testStatistic = c("standard", "wilcoxon"), time.summary.type = c("slope", + "signed.area"), regression.method = c("standard", "ranks"), + return.x = FALSE, knn.neighbors = 10, random.seed = NULL, + nresamp = 20, nresamp.perm = NULL, xl.mode = c("regular", + "firsttime", "next20", "lasttime"), xl.time = NULL, xl.prevfit = NULL) +{ + this.call = match.call() + resp.type.arg = match.arg(resp.type) + assay.type = match.arg(assay.type) + xl.mode = match.arg(xl.mode) + if (!is.null(random.seed)) { + set.seed(random.seed) + } + if (is.null(nresamp.perm)) { + nresamp.perm = nresamp + } + nresamp.perm = min(nresamp, nresamp.perm) + if (xl.mode == "regular" | xl.mode == "firsttime") { + x = NULL + xresamp = NULL + ttstar0 = NULL + evo = NULL + ystar = NULL + sdstar.keep = NULL + censoring.status = NULL + sdstar = NULL + pi0 = NULL + stand.contrasts = NULL + stand.contrasts.star = NULL + stand.contrasts.95 = NULL + foldchange = NULL + foldchange.star = NULL + perms = NULL + permsy = NULL + eigengene = NULL + eigengene.number = NULL + testStatistic <- match.arg(testStatistic) + time.summary.type <- match.arg(time.summary.type) + regression.method <- match.arg(regression.method) + x = data$x + y = data$y + argy = y + if (!is.null(data$eigengene.number)) { + eigengene.number = data$eigengene.number + } + if (sum(is.na(x)) > 0) { + x = impute.knn(x, k = knn.neighbors) + if (!is.matrix(x)) { + x = x$data + } + } + are.blocks.specified = FALSE + cond = (resp.type == "One class") | (resp.type == "Two class unpaired timecourse") | + (resp.type == "One class unpaired timecourse") | + (resp.type == "Two class paired timecourse") | (resp.type == + "Pattern discovery") + if (assay.type == "seq" & cond) { + stop(paste("Resp.type=", resp.type, " not allowed when assay.type='seq'")) + } + if (assay.type == "seq" & min(x) < 0) { + stop(paste("Negative values not allowed when assay.type='seq'")) + } + if (assay.type == "seq" & (sum(x%%1 != 0) != 0)) { + stop("Non-integer values not alled when assay.type='seq'") + } + if (assay.type == "seq" & center.arrays) { + stop(paste("Centering not allowed when assay.type='seq'")) + } + if (assay.type == "seq" & regression.method == "ranks") { + stop(paste("regression.method==ranks not allowed when assay.type='seq'")) + } + if (center.arrays) { + x <- scale(x, center = apply(x, 2, median), scale = FALSE) + } + depth = scaling.factors = rep(NA, ncol(x)) + scaling.factors = (prod(depth)^(1/length(depth)))/depth + if (assay.type == "seq") { + cat("Estimating sequencing depths...", fill = T) + depth = samr.estimate.depth(x) + cat("Resampling to get new data matrices...", fill = T) + xresamp = resa(x, depth, nresamp = nresamp) + } + scaling.factors = (prod(depth)^(1/length(depth)))/depth + if (resp.type == samr.const.twoclass.unpaired.response) { + if (substring(y[1], 2, 6) == "Block" | substring(y[1], + 2, 6) == "block") { + junk = parse.block.labels.for.2classes(y) + y = junk$y + blocky = junk$blocky + are.blocks.specified = TRUE + } + } + if (resp.type == samr.const.twoclass.unpaired.response | + resp.type == samr.const.twoclass.paired.response | + resp.type == samr.const.oneclass.response | resp.type == + samr.const.quantitative.response | resp.type == samr.const.multiclass.response) { + y = as.numeric(y) + } + sd.internal = NULL + if (resp.type == samr.const.twoclass.unpaired.timecourse.response | + resp.type == samr.const.twoclass.paired.timecourse.response | + resp.type == samr.const.oneclass.timecourse.response) { + junk = parse.time.labels.and.summarize.data(x, y, + resp.type, time.summary.type) + y = junk$y + x = junk$x + sd.internal = sqrt(rowMeans(junk$sd^2)) + if (min(table(y)) == 1) { + cat("", fill = T) + cat("Warning: only one timecourse in one or more classes;\nSAM plot and FDRs will be unreliable; only gene scores are informative", + fill = T) + } + } + if (resp.type == samr.const.twoclass.unpaired.timecourse.response) { + resp.type = samr.const.twoclass.unpaired.response + } + if (resp.type == samr.const.twoclass.paired.timecourse.response) { + resp.type = samr.const.twoclass.paired.response + } + if (resp.type == samr.const.oneclass.timecourse.response) { + resp.type = samr.const.oneclass.response + } + stand.contrasts = NULL + stand.contrasts.95 = NULL + if (resp.type == samr.const.survival.response) { + censoring.status = data$censoring.status + } + check.format(y, resp.type = resp.type, censoring.status = censoring.status) + if (resp.type == samr.const.quantitative.response & regression.method == + "ranks") { + y = rank(y) + x = t(apply(x, 1, rank)) + } + n <- nrow(x) + ny <- length(y) + sd <- NULL + numer <- NULL + if (resp.type == samr.const.twoclass.unpaired.response & + testStatistic == "standard" & assay.type == "array") { + init.fit <- ttest.func(x, y, sd = sd.internal) + numer <- init.fit$numer + sd <- init.fit$sd + } + if (resp.type == samr.const.twoclass.unpaired.response & + testStatistic == "wilcoxon" & assay.type == "array") { + init.fit <- wilcoxon.func(x, y) + numer <- init.fit$numer + sd <- init.fit$sd + } + if (resp.type == samr.const.oneclass.response & assay.type == + "array") { + init.fit <- onesample.ttest.func(x, y, sd = sd.internal) + numer <- init.fit$numer + sd <- init.fit$sd + } + if (resp.type == samr.const.twoclass.paired.response & + assay.type == "array") { + init.fit <- paired.ttest.func(x, y, sd = sd.internal) + numer <- init.fit$numer + sd <- init.fit$sd + } + if (resp.type == samr.const.survival.response & assay.type == + "array") { + init.fit <- cox.func(x, y, censoring.status) + numer <- init.fit$numer + sd <- init.fit$sd + } + if (resp.type == samr.const.multiclass.response & assay.type == + "array") { + init.fit <- multiclass.func(x, y) + numer <- init.fit$numer + sd <- init.fit$sd + } + if (resp.type == samr.const.quantitative.response & assay.type == + "array") { + init.fit <- quantitative.func(x, y) + numer <- init.fit$numer + sd <- init.fit$sd + } + if (resp.type == samr.const.patterndiscovery.response & + assay.type == "array") { + init.fit <- patterndiscovery.func(x) + numer <- init.fit$numer + sd <- init.fit$sd + } + if ((resp.type == samr.const.quantitative.response & + (testStatistic == "wilcoxon" | regression.method == + "ranks" & assay.type == "array") | resp.type == + samr.const.patterndiscovery.response) | resp.type == + samr.const.twoclass.unpaired.response & assay.type == + "array" & testStatistic == "wilcoxon" | (nrow(x) < + 500) & is.null(s0) & is.null(s0.perc)) { + s0 = quantile(sd, 0.05) + s0.perc = 0.05 + } + if (is.null(s0) & assay.type == "array") { + if (!is.null(s0.perc)) { + if ((s0.perc != -1 & s0.perc < 0) | s0.perc > + 100) { + stop("Illegal value for s0.perc: must be between 0 and 100, or equal\nto (-1) (meaning that s0 should be set to zero)") + } + if (s0.perc == -1) { + s0 = 0 + } + if (s0.perc >= 0) { + s0 <- quantile(init.fit$sd, s0.perc/100) + } + } + if (is.null(s0.perc)) { + s0 = est.s0(init.fit$tt, init.fit$sd)$s0.hat + s0.perc = 100 * sum(init.fit$sd < s0)/length(init.fit$sd) + } + } + if (assay.type == "seq") { + s0 = 0 + s0.perc = 0 + } + if (resp.type == samr.const.twoclass.unpaired.response & + testStatistic == "standard" & assay.type == "array") { + tt <- ttest.func(x, y, s0 = s0, sd = sd.internal)$tt + } + if (resp.type == samr.const.twoclass.unpaired.response & + testStatistic == "wilcoxon" & assay.type == "array") { + tt <- wilcoxon.func(x, y, s0 = s0)$tt + } + if (resp.type == samr.const.oneclass.response & assay.type == + "array") { + tt <- onesample.ttest.func(x, y, s0 = s0, sd = sd.internal)$tt + } + if (resp.type == samr.const.twoclass.paired.response & + assay.type == "array") { + tt <- paired.ttest.func(x, y, s0 = s0, sd = sd.internal)$tt + } + if (resp.type == samr.const.survival.response & assay.type == + "array") { + tt <- cox.func(x, y, censoring.status, s0 = s0)$tt + } + if (resp.type == samr.const.multiclass.response & assay.type == + "array") { + junk2 <- multiclass.func(x, y, s0 = s0) + tt = junk2$tt + stand.contrasts = junk2$stand.contrasts + } + if (resp.type == samr.const.quantitative.response & assay.type == + "array") { + tt <- quantitative.func(x, y, s0 = s0)$tt + } + if (resp.type == samr.const.patterndiscovery.response & + assay.type == "array") { + junk <- patterndiscovery.func(x, s0 = s0, eigengene.number = eigengene.number) + tt <- junk$tt + eigengene = junk$eigengene + } + if (resp.type == samr.const.twoclass.unpaired.response & + assay.type == "seq") { + junk = wilcoxon.unpaired.seq.func(xresamp, y) + tt = junk$tt + numer = junk$numer + sd = junk$sd + } + if (resp.type == samr.const.twoclass.paired.response & + assay.type == "seq") { + junk <- wilcoxon.paired.seq.func(xresamp, y) + tt = junk$tt + numer = junk$numer + sd = junk$sd + } + if (resp.type == samr.const.quantitative.response & assay.type == + "seq") { + junk <- quantitative.seq.func(xresamp, y) + tt = junk$tt + numer = junk$numer + sd = junk$sd + } + if (resp.type == samr.const.survival.response & assay.type == + "seq") { + junk <- cox.seq.func(xresamp, y, censoring.status) + tt = junk$tt + numer = junk$numer + sd = junk$sd + } + if (resp.type == samr.const.multiclass.response & assay.type == + "seq") { + junk2 <- multiclass.seq.func(xresamp, y) + tt = junk2$tt + numer = junk2$numer + sd = junk2$sd + stand.contrasts = junk2$stand.contrasts + } + if (resp.type == samr.const.quantitative.response | resp.type == + samr.const.multiclass.response | resp.type == samr.const.survival.response) { + junk <- getperms(y, nperms) + perms = junk$perms + all.perms.flag = junk$all.perms.flag + nperms.act = junk$nperms.act + } + if (resp.type == samr.const.twoclass.unpaired.response) { + if (are.blocks.specified) { + junk = compute.block.perms(y, blocky, nperms) + permsy = matrix(junk$permsy, ncol = length(y)) + all.perms.flag = junk$all.perms.flag + nperms.act = junk$nperms.act + } + else { + junk <- getperms(y, nperms) + permsy = matrix(y[junk$perms], ncol = length(y)) + all.perms.flag = junk$all.perms.flag + nperms.act = junk$nperms.act + } + } + if (resp.type == samr.const.oneclass.response) { + if ((length(y) * log(2)) < log(nperms)) { + allii = 0:((2^length(y)) - 1) + nperms.act = 2^length(y) + all.perms.flag = 1 + } + else { + nperms.act = nperms + all.perms.flag = 0 + } + permsy = matrix(NA, nrow = nperms.act, ncol = length(y)) + if (all.perms.flag == 1) { + k = 0 + for (i in allii) { + junk = integer.base.b(i, b = 2) + if (length(junk) < length(y)) { + junk = c(rep(0, length(y) - length(junk)), + junk) + } + k = k + 1 + permsy[k, ] = y * (2 * junk - 1) + } + } + else { + for (i in 1:nperms.act) { + permsy[i, ] = sample(c(-1, 1), size = length(y), + replace = TRUE) + } + } + } + if (resp.type == samr.const.twoclass.paired.response) { + junk = compute.block.perms(y, abs(y), nperms) + permsy = junk$permsy + all.perms.flag = junk$all.perms.flag + nperms.act = junk$nperms.act + } + if (resp.type == samr.const.patterndiscovery.response) { + nperms.act = nperms + perms = NULL + permsy = NULL + all.perms.flag = FALSE + } + sdstar.keep <- NULL + if (assay.type != "seq") { + sdstar.keep <- matrix(0, ncol = nperms.act, nrow = nrow(x)) + } + ttstar <- matrix(0, nrow = nrow(x), ncol = nperms.act) + foldchange.star = NULL + if (resp.type == samr.const.twoclass.unpaired.response | + resp.type == samr.const.twoclass.paired.response) { + foldchange.star <- matrix(0, nrow = nrow(x), ncol = nperms.act) + } + if (resp.type == samr.const.multiclass.response) { + stand.contrasts.star = array(NA, c(nrow(x), length(table(y)), + nperms.act)) + } + } + if (xl.mode == "next20" | xl.mode == "lasttime") { + evo = xl.prevfit$evo + tt = xl.prevfit$tt + numer = xl.prevfit$numer + eigengene = xl.prevfit$eigengene + eigengene.number = xl.prevfit$eigengene.number + sd = xl.prevfit$sd - xl.prevfit$s0 + sd.internal = xl.prevfit$sd.internal + ttstar = xl.prevfit$ttstar + ttstar0 = xl.prevfit$ttstar0 + n = xl.prevfit$n + pi0 = xl.prevfit$pi0 + foldchange = xl.prevfit$foldchange + y = xl.prevfit$y + x = xl.prevfit$x + xresamp = xl.prevfit$xresamp + censoring.status = xl.prevfit$censoring.status + argy = xl.prevfit$argy + testStatistic = xl.prevfit$testStatistic + foldchange.star = xl.prevfit$foldchange.star + s0 = xl.prevfit$s0 + s0.perc = xl.prevfit$s0.perc + resp.type = xl.prevfit$resp.type + resp.type.arg = xl.prevfit$resp.type.arg + assay.type = xl.prevfit$assay.type + sdstar.keep = xl.prevfit$sdstar.keep + resp.type = xl.prevfit$resp.type + stand.contrasts = xl.prevfit$stand.contrasts + stand.contrasts.star = xl.prevfit$stand.contrasts.star + stand.contrasts.95 = xl.prevfit$stand.contrasts.95 + perms = xl.prevfit$perms + permsy = xl.prevfit$permsy + nperms = xl.prevfit$nperms + nperms.act = xl.prevfit$nperms.act + all.perms.flag = xl.prevfit$all.perms.flag + depth = xl.prevfit$depth + scaling.factors = xl.prevfit$scaling.factors + nresamp = xl.prevfit$nresamp + nresamp.perm = xl.prevfit$nresamp.perm + } + if (xl.mode == "regular") { + first = 1 + last = nperms.act + } + if (xl.mode == "firsttime") { + first = 1 + last = 1 + } + if (xl.mode == "next20") { + first = xl.time + last = min(xl.time + 19, nperms.act - 1) + } + if (xl.mode == "lasttime") { + first = nperms.act + last = nperms.act + } + for (b in first:last) { + cat(c("perm=", b), fill = TRUE) + if (assay.type == "array") { + xstar <- x + } + if (assay.type == "seq") { + xstar <- xresamp[, , 1:nresamp.perm] + } + if (resp.type == samr.const.oneclass.response) { + ystar = permsy[b, ] + if (testStatistic == "standard") { + ttstar[, b] <- onesample.ttest.func(xstar, ystar, + s0 = s0, sd = sd.internal)$tt + } + } + if (resp.type == samr.const.twoclass.paired.response) { + ystar = permsy[b, ] + if (assay.type == "array") { + ttstar[, b] <- paired.ttest.func(xstar, ystar, + s0 = s0, sd = sd.internal)$tt + foldchange.star[, b] = foldchange.paired(xstar, + ystar, data$logged2) + } + if (assay.type == "seq") { + ttstar[, b] <- wilcoxon.paired.seq.func(xstar, + ystar)$tt + foldchange.star[, b] <- foldchange.seq.twoclass.paired(x, + ystar, depth) + } + } + if (resp.type == samr.const.twoclass.unpaired.response) { + ystar = permsy[b, ] + if (assay.type == "array") { + if (testStatistic == "standard") { + junk <- ttest.func(xstar, ystar, s0 = s0, sd = sd.internal) + } + if (testStatistic == "wilcoxon") { + junk <- wilcoxon.func(xstar, ystar, s0 = s0) + } + ttstar[, b] <- junk$tt + sdstar.keep[, b] <- junk$sd + foldchange.star[, b] = foldchange.twoclass(xstar, + ystar, data$logged2) + } + if (assay.type == "seq") { + ttstar[, b] <- wilcoxon.unpaired.seq.func(xstar, + ystar)$tt + foldchange.star[, b] <- foldchange.seq.twoclass.unpaired(x, + ystar, depth) + } + } + if (resp.type == samr.const.survival.response) { + o <- perms[b, ] + if (assay.type == "array") { + ttstar[, b] <- cox.func(xstar, y[o], censoring.status = censoring.status[o], + s0 = s0)$tt + } + if (assay.type == "seq") { + ttstar[, b] <- cox.seq.func(xstar, y[o], censoring.status = censoring.status[o])$tt + } + } + if (resp.type == samr.const.multiclass.response) { + ystar = y[perms[b, ]] + if (assay.type == "array") { + junk <- multiclass.func(xstar, ystar, s0 = s0) + ttstar[, b] <- junk$tt + sdstar.keep[, b] <- junk$sd + stand.contrasts.star[, , b] = junk$stand.contrasts + } + if (assay.type == "seq") { + junk <- multiclass.seq.func(xstar, ystar) + ttstar[, b] <- junk$tt + stand.contrasts.star[, , b] <- junk$stand.contrasts + } + } + if (resp.type == samr.const.quantitative.response) { + ystar = y[perms[b, ]] + if (assay.type == "array") { + junk <- quantitative.func(xstar, ystar, s0 = s0) + ttstar[, b] <- junk$tt + sdstar.keep[, b] <- junk$sd + } + if (assay.type == "seq") { + junk <- quantitative.seq.func(xstar, ystar) + ttstar[, b] <- junk$tt + } + } + if (resp.type == samr.const.patterndiscovery.response) { + xstar = permute.rows(x) + junk <- patterndiscovery.func(xstar, s0 = s0, eigengene.number = eigengene.number) + ttstar[, b] <- junk$tt + sdstar.keep[, b] <- junk$sd + } + } + if (xl.mode == "regular" | xl.mode == "lasttime") { + ttstar0 <- ttstar + for (j in 1:ncol(ttstar)) { + ttstar[, j] <- -1 * sort(-1 * ttstar[, j]) + } + for (i in 1:nrow(ttstar)) { + ttstar[i, ] <- sort(ttstar[i, ]) + } + evo <- apply(ttstar, 1, mean) + evo <- evo[length(evo):1] + sdstar <- sdstar.keep + pi0 = 1 + if (resp.type != samr.const.multiclass.response) { + qq <- quantile(ttstar, c(0.25, 0.75)) + } + if (resp.type == samr.const.multiclass.response) { + qq <- quantile(ttstar, c(0, 0.5)) + } + pi0 <- sum(tt > qq[1] & tt < qq[2])/(0.5 * length(tt)) + foldchange = NULL + if (resp.type == samr.const.twoclass.unpaired.response & + assay.type == "array") { + foldchange = foldchange.twoclass(x, y, data$logged2) + } + if (resp.type == samr.const.twoclass.paired.response & + assay.type == "array") { + foldchange = foldchange.paired(x, y, data$logged2) + } + if (resp.type == samr.const.oneclass.response & assay.type == + "array") { + } + stand.contrasts.95 = NULL + if (resp.type == samr.const.multiclass.response) { + stand.contrasts.95 = quantile(stand.contrasts.star, + c(0.025, 0.975)) + } + if (resp.type == samr.const.twoclass.unpaired.response & + assay.type == "seq") { + foldchange <- foldchange.seq.twoclass.unpaired(x, + y, depth) + } + if (resp.type == samr.const.twoclass.paired.response & + assay.type == "seq") { + foldchange <- foldchange.seq.twoclass.paired(x, y, + depth) + } + if (return.x == FALSE) { + x = NULL + } + } + return(list(n = n, x = x, xresamp = xresamp, y = y, argy = argy, + censoring.status = censoring.status, testStatistic = testStatistic, + nperms = nperms, nperms.act = nperms.act, tt = tt, numer = numer, + sd = sd + s0, sd.internal = sd.internal, s0 = s0, s0.perc = s0.perc, + evo = evo, perms = perms, permsy = permsy, nresamp = nresamp, + nresamp.perm = nresamp.perm, all.perms.flag = all.perms.flag, + ttstar = ttstar, ttstar0 = ttstar0, eigengene = eigengene, + eigengene.number = eigengene.number, pi0 = pi0, foldchange = foldchange, + foldchange.star = foldchange.star, sdstar.keep = sdstar.keep, + resp.type = resp.type, resp.type.arg = resp.type.arg, + assay.type = assay.type, stand.contrasts = stand.contrasts, + stand.contrasts.star = stand.contrasts.star, stand.contrasts.95 = stand.contrasts.95, + depth = depth, call = this.call)) +} + +#' @title Estimate sequencing depths +#' @param x data matrix. nrow=#gene, ncol=#sample +#' @return depth: estimated sequencing depth. a vector with len sample. +samr.estimate.depth <- function(x) { + iter <- 5 + cmeans <- colSums(x)/sum(x) + for (i in 1:iter) { + n0 <- rowSums(x) %*% t(cmeans) + prop <- rowSums((x - n0)^2/(n0 + 1e-08)) + qs <- quantile(prop, c(0.25, 0.75)) + keep <- (prop >= qs[1]) & (prop <= qs[2]) + cmeans <- colMeans(x[keep, ]) + cmeans <- cmeans/sum(cmeans) + } + depth <- cmeans/mean(cmeans) + return(depth) +} + +#' @title Resampling +#' @param x data matrix. nrow=#gene, ncol=#sample +#' @param d estimated sequencing depth +#' @param nresamp number of resamplings +#' @return xresamp: an rank array with dim #gene*#sample*nresamp +#' @description Corresponds to `samr::resample` +resa <- function(x, d, nresamp = 20) { + ng <- nrow(x) + ns <- ncol(x) + dbar <- exp(mean(log(d))) + xresamp <- array(0, dim = c(ng, ns, nresamp)) + for (k in 1:nresamp) { + for (j in 1:ns) { + xresamp[, j, k] <- rpois(n = ng, lambda = (dbar/d[j]) * + x[, j]) + runif(ng) * 0.1 + } + } + for (k in 1:nresamp) { + xresamp[, , k] <- t(rankcols(t(xresamp[, , k]))) + } + return(xresamp) +} + +#' @title Rank columns +#' @description Ranks the elements within each col of the matrix x and returns +#' these ranks in a matrix +#' @note this function is equivalent to `samr::rankcol`, but uses `apply` to +#' rank the colums instead of a compiled Fortran function which was causing our +#' DEGanalysis functions to freeze in large datasets. +rankcols <- function(x) { + # ranks the elements within each col of the matrix x + # and returns these ranks in a matrix + n = nrow(x) + p = ncol(x) + mode(n) = "integer" + mode(p) = "integer" + mode(x) = "double" + xr <- apply(x, 2, rank) + return(xr) +} + +#' @title Check format +check.format = function(y, resp.type, censoring.status = NULL) { + # here i do some format checks for the input data$y + # note that checks for time course data are done in the + # parse function for time course; + # we then check the output from the parser in this function + if (resp.type == samr.const.twoclass.unpaired.response | + resp.type == samr.const.twoclass.unpaired.timecourse.response) { + if (sum(y == 1) + sum(y == 2) != length(y)) { + stop(paste("Error in input response data: response type ", + resp.type, " specified; values must be 1 or 2")) + } + } + if (resp.type == samr.const.twoclass.paired.response | resp.type == + samr.const.twoclass.paired.timecourse.response) { + if (sum(y) != 0) { + stop(paste("Error in input response data: response type ", + resp.type, " specified; values must be -1, 1, -2, 2, etc")) + } + if (sum(table(y[y > 0]) != abs(table(y[y < 0])))) { + stop(paste("Error in input response data: response type ", + resp.type, " specified; values must be -1, 1, -2, 2, etc")) + } + } + if (resp.type == samr.const.oneclass.response | resp.type == + samr.const.oneclass.timecourse.response) { + if (sum(y == 1) != length(y)) { + stop(paste("Error in input response data: response type ", + resp.type, " specified; values must all be 1")) + } + } + if (resp.type == samr.const.multiclass.response) { + tt = table(y) + nc = length(tt) + if (sum(y <= nc & y > 0) < length(y)) { + stop(paste("Error in input response data: response type ", + resp.type, " specified; values must be 1,2, ... number of classes")) + } + for (k in 1:nc) { + if (sum(y == k) < 2) { + stop(paste("Error in input response data: response type ", + resp.type, " specified; there must be >1 sample per class")) + } + } + } + if (resp.type == samr.const.quantitative.response) { + if (!is.numeric(y)) { + stop(paste("Error in input response data: response type", + resp.type, " specified; values must be numeric")) + } + } + if (resp.type == samr.const.survival.response) { + if (is.null(censoring.status)) { + stop(paste("Error in input response data: response type ", + resp.type, " specified; error in censoring indicator")) + } + if (!is.numeric(y) | sum(y < 0) > 0) { + stop(paste("Error in input response data: response type ", + resp.type, " specified; survival times must be numeric and nonnegative")) + if (sum(censoring.status == 0) + sum(censoring.status == + 1) != length(censoring.status)) { + stop(paste("Error in input response data: response type ", + resp.type, " specified; censoring indicators must be 0 (censored) or 1 (failed)")) + } + } + if (sum(censoring.status == 1) < 1) { + stop(paste("Error in input response data: response type ", + resp.type, " specified; there are no uncensored observations")) + } + } + return() +} + +#' @title Twoclass Wilcoxon statistics +#' @param xresamp an rank array with dim #gene*#sample*nresamp +#' @param y outcome vector of values 1 and 2 +#' @return the statistic. +wilcoxon.unpaired.seq.func <- function(xresamp, y) { + tt <- rep(0, dim(xresamp)[1]) + for (i in 1:dim(xresamp)[3]) { + tt <- tt + rowSums(xresamp[, y == 2, i]) - sum(y == 2) * + (length(y) + 1)/2 + } + tt <- tt/dim(xresamp)[3] + return(list(tt = tt, numer = tt, sd = rep(1, length(tt)))) +} + +getperms = function(y, nperms) { + total.perms = factorial(length(y)) + if (total.perms <= nperms) { + perms = permute(1:length(y)) + all.perms.flag = 1 + nperms.act = total.perms + } + if (total.perms > nperms) { + perms = matrix(NA, nrow = nperms, ncol = length(y)) + for (i in 1:nperms) { + perms[i, ] = sample(1:length(y), size = length(y)) + } + all.perms.flag = 0 + nperms.act = nperms + } + return(list(perms = perms, all.perms.flag = all.perms.flag, + nperms.act = nperms.act)) +} + +#' @title Foldchange of twoclass unpaired sequencing data +foldchange.seq.twoclass.unpaired <- function(x, y, depth) +{ + x.norm <- scale(x, center = F, scale = depth) + 1e-08 + fc <- rowMedians(x.norm[, y == 2])/rowMedians(x.norm[, y == + 1]) + return(fc) +} diff --git a/man/check.format.Rd b/man/check.format.Rd new file mode 100644 index 0000000..dcee24c --- /dev/null +++ b/man/check.format.Rd @@ -0,0 +1,11 @@ +% Generated by roxygen2: do not edit by hand +% Please edit documentation in R/samr.R +\name{check.format} +\alias{check.format} +\title{Check format} +\usage{ +check.format(y, resp.type, censoring.status = NULL) +} +\description{ +Check format +} diff --git a/man/foldchange.seq.twoclass.unpaired.Rd b/man/foldchange.seq.twoclass.unpaired.Rd new file mode 100644 index 0000000..b95217e --- /dev/null +++ b/man/foldchange.seq.twoclass.unpaired.Rd @@ -0,0 +1,11 @@ +% Generated by roxygen2: do not edit by hand +% Please edit documentation in R/samr.R +\name{foldchange.seq.twoclass.unpaired} +\alias{foldchange.seq.twoclass.unpaired} +\title{Foldchange of twoclass unpaired sequencing data} +\usage{ +foldchange.seq.twoclass.unpaired(x, y, depth) +} +\description{ +Foldchange of twoclass unpaired sequencing data +} diff --git a/man/rankcols.Rd b/man/rankcols.Rd new file mode 100644 index 0000000..09d9e8b --- /dev/null +++ b/man/rankcols.Rd @@ -0,0 +1,17 @@ +% Generated by roxygen2: do not edit by hand +% Please edit documentation in R/samr.R +\name{rankcols} +\alias{rankcols} +\title{Rank columns} +\usage{ +rankcols(x) +} +\description{ +Ranks the elements within each col of the matrix x and returns +these ranks in a matrix +} +\note{ +this function is equivalent to `samr::rankcol`, but uses `apply` to +rank the colums instead of a compiled Fortran function which was causing our +DEGanalysis functions to freeze in large datasets. +} diff --git a/man/resa.Rd b/man/resa.Rd new file mode 100644 index 0000000..74b9d5f --- /dev/null +++ b/man/resa.Rd @@ -0,0 +1,21 @@ +% Generated by roxygen2: do not edit by hand +% Please edit documentation in R/samr.R +\name{resa} +\alias{resa} +\title{Resampling} +\usage{ +resa(x, d, nresamp = 20) +} +\arguments{ +\item{x}{data matrix. nrow=#gene, ncol=#sample} + +\item{d}{estimated sequencing depth} + +\item{nresamp}{number of resamplings} +} +\value{ +xresamp: an rank array with dim #gene*#sample*nresamp +} +\description{ +Corresponds to `samr::resample` +} diff --git a/man/sammy.Rd b/man/sammy.Rd new file mode 100644 index 0000000..a3d45e7 --- /dev/null +++ b/man/sammy.Rd @@ -0,0 +1,32 @@ +% Generated by roxygen2: do not edit by hand +% Please edit documentation in R/samr.R +\name{sammy} +\alias{sammy} +\title{Significance analysis of microarrays} +\usage{ +sammy( + data, + resp.type = c("Quantitative", "Two class unpaired", "Survival", "Multiclass", + "One class", "Two class paired", "Two class unpaired timecourse", + "One class timecourse", "Two class paired timecourse", "Pattern discovery"), + assay.type = c("array", "seq"), + s0 = NULL, + s0.perc = NULL, + nperms = 100, + center.arrays = FALSE, + testStatistic = c("standard", "wilcoxon"), + time.summary.type = c("slope", "signed.area"), + regression.method = c("standard", "ranks"), + return.x = FALSE, + knn.neighbors = 10, + random.seed = NULL, + nresamp = 20, + nresamp.perm = NULL, + xl.mode = c("regular", "firsttime", "next20", "lasttime"), + xl.time = NULL, + xl.prevfit = NULL +) +} +\description{ +This function is an adaptation of `samr::samr` +} diff --git a/man/samr.estimate.depth.Rd b/man/samr.estimate.depth.Rd new file mode 100644 index 0000000..f91d5e5 --- /dev/null +++ b/man/samr.estimate.depth.Rd @@ -0,0 +1,17 @@ +% Generated by roxygen2: do not edit by hand +% Please edit documentation in R/samr.R +\name{samr.estimate.depth} +\alias{samr.estimate.depth} +\title{Estimate sequencing depths} +\usage{ +samr.estimate.depth(x) +} +\arguments{ +\item{x}{data matrix. nrow=#gene, ncol=#sample} +} +\value{ +depth: estimated sequencing depth. a vector with len sample. +} +\description{ +Estimate sequencing depths +} diff --git a/man/wilcoxon.unpaired.seq.func.Rd b/man/wilcoxon.unpaired.seq.func.Rd new file mode 100644 index 0000000..1925ecc --- /dev/null +++ b/man/wilcoxon.unpaired.seq.func.Rd @@ -0,0 +1,19 @@ +% Generated by roxygen2: do not edit by hand +% Please edit documentation in R/samr.R +\name{wilcoxon.unpaired.seq.func} +\alias{wilcoxon.unpaired.seq.func} +\title{Twoclass Wilcoxon statistics} +\usage{ +wilcoxon.unpaired.seq.func(xresamp, y) +} +\arguments{ +\item{xresamp}{an rank array with dim #gene*#sample*nresamp} + +\item{y}{outcome vector of values 1 and 2} +} +\value{ +the statistic. +} +\description{ +Twoclass Wilcoxon statistics +} From 69051ff1029c04d16068708a39256346b0fd329a Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Tue, 26 May 2020 12:00:56 +0200 Subject: [PATCH 05/99] Fixed documentation --- DESCRIPTION | 4 +- NAMESPACE | 4 + R/DIscBIO-generic-DEGanalysis.R | 9 +- R/DIscBIO-generic-DEGanalysis2clust.R | 4 +- R/samr-adapted.R | 606 +++++++++++++++++++++++- man/check.format.Rd | 9 +- man/foldchange.seq.twoclass.unpaired.Rd | 9 +- man/rankcols.Rd | 5 +- man/resa.Rd | 2 +- man/sammy.Rd | 39 +- man/samr.estimate.depth.Rd | 2 +- man/wilcoxon.unpaired.seq.func.Rd | 2 +- 12 files changed, 676 insertions(+), 19 deletions(-) diff --git a/DESCRIPTION b/DESCRIPTION index 03af8d9..8937ae7 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -33,7 +33,7 @@ Encoding: UTF-8 Imports: methods, TSCAN, boot, httr, mclust, statmod, biomaRt, samr, igraph, RWeka, partykit, grid, philentropy, NetIndices, png, matrixStats, grDevices, readr, RColorBrewer, ggplot2, rpart, fpc, cluster, rpart.plot, amap, dplyr, - tsne, AnnotationDbi, org.Hs.eg.db, calibrate, graphics, stats, utils + tsne, AnnotationDbi, org.Hs.eg.db, calibrate, graphics, stats, utils, impute Depends: R (>= 4.0), SingleCellExperiment Suggests: @@ -94,4 +94,4 @@ Collate: 'customConverters.R' 'datasets.R' 'retrieveBiomart.R' - 'samr.R' + 'samr-adapted.R' diff --git a/NAMESPACE b/NAMESPACE index 838c1d0..33526ff 100644 --- a/NAMESPACE +++ b/NAMESPACE @@ -127,6 +127,8 @@ importFrom(igraph,graph.adjacency) importFrom(igraph,graph.data.frame) importFrom(igraph,mean_distance) importFrom(igraph,minimum.spanning.tree) +importFrom(impute,impute.knn) +importFrom(matrixStats,rowMedians) importFrom(matrixStats,rowVars) importFrom(mclust,Mclust) importFrom(mclust,mclustBIC) @@ -161,6 +163,8 @@ importFrom(stats,pnbinom) importFrom(stats,prcomp) importFrom(stats,predict) importFrom(stats,quantile) +importFrom(stats,rpois) +importFrom(stats,runif) importFrom(stats,var) importFrom(tsne,tsne) importFrom(utils,capture.output) diff --git a/R/DIscBIO-generic-DEGanalysis.R b/R/DIscBIO-generic-DEGanalysis.R index 404c559..c7803c1 100644 --- a/R/DIscBIO-generic-DEGanalysis.R +++ b/R/DIscBIO-generic-DEGanalysis.R @@ -195,13 +195,12 @@ setMethod( o <- c(1:K) oo <- o[-length(o)] com <- sum(oo) - if (!quiet) - cat("Number of comparisons: ", com * 2, "\n") + if (!quiet) cat("Number of comparisons: ", com * 2, "\n") comNUM <- paste("comp", c(1:com), sep = "") DEGsTable <- data.frame() DEGsE <- c() DEGsS <- c() - for (i in 1:com) { + for (i in seq_len(com)) { FinalDEGsL <- data.frame() FinalDEGsU <- data.frame() FDRl <- c() @@ -219,7 +218,7 @@ setMethod( data <- list(x = x, y = y, geneid = gname) if (quiet) { invisible(capture.output( - samr.obj <- samr( + samr.obj <- sammy( data, resp.type = "Two class unpaired", assay.type = "seq", @@ -229,7 +228,7 @@ setMethod( ) )) } else { - samr.obj <- samr( + samr.obj <- sammy( data, resp.type = "Two class unpaired", assay.type = "seq", diff --git a/R/DIscBIO-generic-DEGanalysis2clust.R b/R/DIscBIO-generic-DEGanalysis2clust.R index 317df66..18c01af 100644 --- a/R/DIscBIO-generic-DEGanalysis2clust.R +++ b/R/DIscBIO-generic-DEGanalysis2clust.R @@ -105,7 +105,7 @@ setMethod( data = list(x = x, y = y, geneid = gname) if (quiet) { invisible(capture.output({ - samr.obj <- samr( + samr.obj <- sammy( data, resp.type = "Two class unpaired", assay.type = "seq", @@ -116,7 +116,7 @@ setMethod( delta.table <- samr.compute.delta.table(samr.obj) })) } else { - samr.obj <- samr( + samr.obj <- sammy( data, resp.type = "Two class unpaired", assay.type = "seq", diff --git a/R/samr-adapted.R b/R/samr-adapted.R index d5151a5..eebb5d1 100644 --- a/R/samr-adapted.R +++ b/R/samr-adapted.R @@ -20,6 +20,25 @@ samr.const.patterndiscovery.response <- "Pattern discovery" #' @title Significance analysis of microarrays #' @description This function is an adaptation of `samr::samr` +#' @param data Data object with components x- p by n matrix of features, one observation per column (missing values allowed); y- n-vector of outcome measurements; censoring.status- n-vector of censoring censoring.status (1= died or event occurred, 0=survived, or event was censored), needed for a censored survival outcome +#' @param resp.type Problem type: "Quantitative" for a continuous parameter (Available for both array and sequencing data); "Two class unpaired" (for both array and sequencing data); "Survival" for censored survival outcome (for both array and sequencingdata); "Multiclass": more than 2 groups (for both array and sequencing data); "One class" for a single group (only for array data); "Two class paired" for two classes with paired observations (for both array and sequencing data); "Two class unpaired timecourse" (only for array data), "One class time course" (only for array data), "Two class.paired timecourse" (only for array data), or "Pattern discovery" (only for array data) +#' @param assay.type Assay type: "array" for microarray data, "seq" for counts from sequencing +#' @param s0 Exchangeability factor for denominator of test statistic; Default is automatic choice. Only used for array data. +#' @param s0.perc Percentile of standard deviation values to use for s0; default is automatic choice; -1 means s0=0 (different from s0.perc=0, meaning s0=zeroeth percentile of standard deviation values= min of sd values. Only used for array data. +#' @param nperms Number of permutations used to estimate false discovery rates +#' @param center.arrays Should the data for each sample (array) be median centered at the outset? Default =FALSE. Only used for array data. +#' @param testStatistic Test statistic to use in two class unpaired case.Either "standard" (t-statistic) or ,"wilcoxon" (Two-sample wilcoxon or Mann-Whitney test). Only used for array data. +#' @param time.summary.type Summary measure for each time course: "slope", or"signed.area"). Only used for array data. +#' @param regression.method Regression method for quantitative case: "standard",(linear least squares) or "ranks" (linear least squares on ranked data). Only used for array data. +#' @param return.x Should the matrix of feature values be returned? Only useful for time course data, where x contains summaries of the features over time. Otherwise x is the same as the input data data\$x +#' @param knn.neighbors Number of nearest neighbors to use for imputation of missing features values. Only used for array data. +#' @param random.seed Optional initial seed for random number generator (integer) +#' @param nresamp For assay.type="seq", number of resamples used to construct test statistic. Default 20. Only used for sequencing data. +#' @param nresamp.perm For assay.type="seq", number of resamples used to construct test statistic for permutations. Default is equal to nresamp and it must be at most nresamp. Only used for sequencing data. +#' @param xl.mode Used by Excel interface +#' @param xl.time Used by Excel interface +#' @param xl.prevfit Used by Excel interface +#' @importFrom impute impute.knn sammy <- function (data, resp.type = c("Quantitative", "Two class unpaired", "Survival", "Multiclass", "One class", "Two class paired", "Two class unpaired timecourse", "One class timecourse", @@ -631,6 +650,7 @@ samr.estimate.depth <- function(x) { #' @param nresamp number of resamplings #' @return xresamp: an rank array with dim #gene*#sample*nresamp #' @description Corresponds to `samr::resample` +#' @importFrom stats rpois runif resa <- function(x, d, nresamp = 20) { ng <- nrow(x) ns <- ncol(x) @@ -654,6 +674,7 @@ resa <- function(x, d, nresamp = 20) { #' @note this function is equivalent to `samr::rankcol`, but uses `apply` to #' rank the colums instead of a compiled Fortran function which was causing our #' DEGanalysis functions to freeze in large datasets. +#' @param x x rankcols <- function(x) { # ranks the elements within each col of the matrix x # and returns these ranks in a matrix @@ -667,7 +688,10 @@ rankcols <- function(x) { } #' @title Check format -check.format = function(y, resp.type, censoring.status = NULL) { +#' @param y y +#' @param resp.type resp type +#' @param censoring.status censoring status +check.format <- function(y, resp.type, censoring.status = NULL) { # here i do some format checks for the input data$y # note that checks for time course data are done in the # parse function for time course; @@ -752,7 +776,15 @@ wilcoxon.unpaired.seq.func <- function(xresamp, y) { tt <- tt/dim(xresamp)[3] return(list(tt = tt, numer = tt, sd = rep(1, length(tt)))) } - +wilcoxon.paired.seq.func <- function(xresamp, y) { + tt <- rep(0, dim(xresamp)[1]) + for (i in 1:dim(xresamp)[3]) { + tt <- tt + rowSums(xresamp[, y > 0, i]) - sum(y > 0) * + (length(y) + 1)/2 + } + tt <- tt/dim(xresamp)[3] + return(list(tt = tt, numer = tt, sd = rep(1, length(tt)))) +} getperms = function(y, nperms) { total.perms = factorial(length(y)) if (total.perms <= nperms) { @@ -771,8 +803,352 @@ getperms = function(y, nperms) { return(list(perms = perms, all.perms.flag = all.perms.flag, nperms.act = nperms.act)) } - +foldchange.twoclass = function(x, y, logged2) { + # if(logged2){x=2^x} + m1 <- rowMeans(x[, y == 1, drop = F]) + m2 <- rowMeans(x[, y == 2, drop = F]) + if (!logged2) { + fc = m2/m1 + } + if (logged2) { + fc = 2^{ + m2 - m1 + } + } + return(fc) +} #' @title Foldchange of twoclass unpaired sequencing data +#' @importFrom matrixStats rowMedians +#' @param x x +#' @param y y +#' @param depth depth +foldchange.seq.twoclass.unpaired <- function(x, y, depth) +{ + x.norm <- scale(x, center = F, scale = depth) + 1e-08 + fc <- rowMedians(x.norm[, y == 2])/rowMedians(x.norm[, y == + 1]) + return(fc) +} +foldchange.seq.twoclass.paired <- function(x, y, depth) { + nc <- ncol(x)/2 + o1 <- o2 <- rep(0, nc) + for (j in 1:nc) { + o1[j] <- which(y == -j) + o2[j] <- which(y == j) + } + x.norm <- scale(x, center = F, scale = depth) + 1e-08 + d <- x.norm[, o2, drop = F]/x.norm[, o1, drop = F] + fc <- rowMedians(d, na.rm = T) + return(fc) +} +permute <- function(elem) { + # generates all perms of the vector elem + if (!missing(elem)) { + if (length(elem) == 2) + return(matrix(c(elem, elem[2], elem[1]), nrow = 2)) + last.matrix <- permute(elem[-1]) + dim.last <- dim(last.matrix) + new.matrix <- matrix(0, nrow = dim.last[1] * (dim.last[2] + + 1), ncol = dim.last[2] + 1) + for (row in 1:(dim.last[1])) { + for (col in 1:(dim.last[2] + 1)) new.matrix[row + + (col - 1) * dim.last[1], ] <- insert.value(last.matrix[row, + ], elem[1], col) + } + return(new.matrix) + } + else cat("Usage: permute(elem)\n\twhere elem is a vector\n") +} +insert.value <- function(vec, newval, pos) { + if (pos == 1) + return(c(newval, vec)) + lvec <- length(vec) + if (pos > lvec) + return(c(vec, newval)) + return(c(vec[1:pos - 1], newval, vec[pos:lvec])) +} +parse.block.labels.for.2classes = function(y) { + #this only works for 2 class case- having form jBlockn, + # where j=1 or 2 + n = length(y) + y.act = rep(NA, n) + blocky = rep(NA, n) + for (i in 1:n) { + blocky[i] = as.numeric(substring(y[i], 7, nchar(y[i]))) + y.act[i] = as.numeric(substring(y[i], 1, 1)) + } + return(list(y.act = y.act, blocky = blocky)) +} +parse.time.labels.and.summarize.data = function(x, + y, resp.type, time.summary.type) { + # parse time labels, and summarize time data for each + # person, via a slope or area + # does some error checking too + n = length(y) + last5char = rep(NA, n) + last3char = rep(NA, n) + for (i in 1:n) { + last3char[i] = substring(y[i], nchar(y[i]) - 2, nchar(y[i])) + last5char[i] = substring(y[i], nchar(y[i]) - 4, nchar(y[i])) + } + if (sum(last3char == "End") != sum(last5char == "Start")) { + stop("Error in format of time course data: a Start or End tag is missing") + } + y.act = rep(NA, n) + timey = rep(NA, n) + person.id = rep(NA, n) + k = 1 + end.flag = FALSE + person.id[1] = 1 + if (substring(y[1], nchar(y[1]) - 4, nchar(y[1])) != "Start") { + stop("Error in format of time course data: first cell should have a Start tag") + } + for (i in 1:n) { + cat(i) + j = 1 + while (substring(y[i], j, j) != "T") { + j = j + 1 + } + end.of.y = j - 1 + y.act[i] = as.numeric(substring(y[i], 1, end.of.y)) + timey[i] = substring(y[i], end.of.y + 5, nchar(y[i])) + if (nchar(timey[i]) > 3 & substring(timey[i], nchar(timey[i]) - + 2, nchar(timey[i])) == "End") { + end.flag = TRUE + timey[i] = substring(timey[i], 1, nchar(timey[i]) - + 3) + } + if (nchar(timey[i]) > 3 & substring(timey[i], nchar(timey[i]) - + 4, nchar(timey[i])) == "Start") { + timey[i] = substring(timey[i], 1, nchar(timey[i]) - + 5) + } + if (i < n & !end.flag) { + person.id[i + 1] = k + } + if (i < n & end.flag) { + k = k + 1 + person.id[i + 1] = k + } + end.flag = FALSE + } + timey = as.numeric(timey) + # do a check that the format was correct + tt = table(person.id, y.act) + junk = function(x) { + sum(x != 0) + } + if (sum(apply(tt, 1, junk) != 1) > 0) { + num = (1:nrow(tt))[apply(tt, 1, junk) > 1] + stop(paste("Error in format of time course data, timecourse #", + as.character(num))) + } + npeople = length(unique(person.id)) + newx = matrix(NA, nrow = nrow(x), ncol = npeople) + sd = matrix(NA, nrow = nrow(x), ncol = npeople) + for (j in 1:npeople) { + jj = person.id == j + tim = timey[jj] + xc = t(scale(t(x[, jj, drop = F]), center = TRUE, scale = FALSE)) + if (time.summary.type == "slope") { + junk = quantitative.func(xc, tim - mean(tim)) + newx[, j] = junk$numer + sd[, j] = junk$sd + } + if (time.summary.type == "signed.area") { + junk = timearea.func(x[, jj, drop = F], tim) + newx[, j] = junk$numer + sd[, j] = junk$sd + } + } + y.unique = y.act[!duplicated(person.id)] + return(list(y = y.unique, x = newx, sd = sd)) +} +ttest.func <- function(x, y, s0 = 0, sd = NULL) { + n1 <- sum(y == 1) + n2 <- sum(y == 2) + p <- nrow(x) + m1 <- rowMeans(x[, y == 1, drop = F]) + m2 <- rowMeans(x[, y == 2, drop = F]) + if (is.null(sd)) { + sd <- sqrt(((n2 - 1) * varr(x[, y == 2], meanx = m2) + + (n1 - 1) * varr(x[, y == 1], meanx = m1)) * (1/n1 + + 1/n2)/(n1 + n2 - 2)) + } + numer <- m2 - m1 + dif.obs <- (numer)/(sd + s0) + return(list(tt = dif.obs, numer = numer, sd = sd)) +} + +wilcoxon.func <- function(x, y, s0 = 0) { + n1 <- sum(y == 1) + n2 <- sum(y == 2) + p = nrow(x) + r2 = rowSums(t(apply(x, 1, rank))[, y == 2, drop = F]) + numer = r2 - (n2/2) * (n2 + 1) - (n1 * n2)/2 + sd = sqrt(n1 * n2 * (n1 + n2 + 1)/12) + tt = (numer)/(sd + s0) + return(list(tt = tt, numer = numer, sd = rep(sd, p))) +} + +onesample.ttest.func <- function(x, y, s0 = 0, sd = NULL) { + n <- length(y) + x <- x * matrix(y, nrow = nrow(x), ncol = ncol(x), byrow = TRUE) + m <- rowMeans(x) + if (is.null(sd)) { + sd <- sqrt(varr(x, meanx = m)/n) + } + dif.obs <- m/(sd + s0) + return(list(tt = dif.obs, numer = m, sd = sd)) +} + +patterndiscovery.func = function(x, s0 = 0, eigengene.number = 1) { + a = mysvd(x, n.components = eigengene.number) + v = a$v[, eigengene.number] + # here we try to guess the most interpretable orientation + # for the eigengene + om = abs(a$u[, eigengene.number]) > quantile(abs(a$u[, eigengene.number]), + 0.95) + if (median(a$u[om, eigengene.number]) < 0) { + v = -1 * v + } + aa = quantitative.func(x, v, s0 = s0) + eigengene = cbind(1:nrow(a$v), v) + dimnames(eigengene) = list(NULL, c("sample number", "value")) + return(list(tt = aa$tt, numer = aa$numer, sd = aa$sd, eigengene = eigengene)) +} + +paired.ttest.func <- function(x, y, s0 = 0, sd = NULL) { + nc <- ncol(x)/2 + o <- 1:nc + o1 <- rep(0, ncol(x)/2) + o2 <- o1 + for (j in 1:nc) { + o1[j] <- (1:ncol(x))[y == -o[j]] + } + for (j in 1:nc) { + o2[j] <- (1:ncol(x))[y == o[j]] + } + d <- x[, o2, drop = F] - x[, o1, drop = F] + su <- x[, o2, drop = F] + x[, o1, drop = F] + if (is.matrix(d)) { + m <- rowMeans(d) + } + if (!is.matrix(d)) { + m <- mean(d) + } + if (is.null(sd)) { + if (is.matrix(d)) { + sd <- sqrt(varr(d, meanx = m)/nc) + } + if (!is.matrix(d)) { + sd <- sqrt(var(d)/nc) + } + } + dif.obs <- m/(sd + s0) + return(list(tt = dif.obs, numer = m, sd = sd)) +} + +cox.func <- function(x, y, censoring.status, s0 = 0) { + # find the index matrix + Dn <- sum(censoring.status == 1) + Dset <- c(1:ncol(x))[censoring.status == 1] # the set of observed + ind <- matrix(0, ncol(x), Dn) + # get the matrix + for (i in 1:Dn) { + ind[y > y[Dset[i]] - 1e-08, i] <- 1/sum(y > y[Dset[i]] - + 1e-08) + } + ind.sums <- rowSums(ind) + x.ind <- x %*% ind + # get the derivatives + numer <- x %*% (censoring.status - ind.sums) + sd <- sqrt((x * x) %*% ind.sums - rowSums(x.ind * x.ind)) + tt <- numer/(sd + s0) + return(list(tt = tt, numer = numer, sd = sd)) +} + +multiclass.func <- function(x, y, s0 = 0) { + ##assumes y is coded 1,2... + nn <- table(y) + m <- matrix(0, nrow = nrow(x), ncol = length(nn)) + v <- m + for (j in 1:length(nn)) { + m[, j] <- rowMeans(x[, y == j]) + v[, j] <- (nn[j] - 1) * varr(x[, y == j], meanx = m[, + j]) + } + mbar <- rowMeans(x) + mm <- m - matrix(mbar, nrow = length(mbar), ncol = length(nn)) + fac <- (sum(nn)/prod(nn)) + scor <- sqrt(fac * (apply(matrix(nn, nrow = nrow(m), ncol = ncol(m), + byrow = TRUE) * mm * mm, 1, sum))) + sd <- sqrt(rowSums(v) * (1/sum(nn - 1)) * sum(1/nn)) + tt <- scor/(sd + s0) + mm.stand = t(scale(t(mm), center = FALSE, scale = sd)) + return(list(tt = tt, numer = scor, sd = sd, stand.contrasts = mm.stand)) +} + +est.s0 <- function(tt, sd, s0.perc = seq(0, 1, by = 0.05)) { + ## estimate s0 (exchangeability) factor for denominator. + ## returns the actual estimate s0 (not a percentile) + br = unique(quantile(sd, seq(0, 1, len = 101))) + nbr = length(br) + a <- cut(sd, br, labels = F) + a[is.na(a)] <- 1 + cv.sd <- rep(0, length(s0.perc)) + for (j in 1:length(s0.perc)) { + w <- quantile(sd, s0.perc[j]) + w[j == 1] <- 0 + tt2 <- tt * sd/(sd + w) + tt2[tt2 == Inf] = NA + sds <- rep(0, nbr - 1) + for (i in 1:(nbr - 1)) { + sds[i] <- stats::mad(tt2[a == i], na.rm = TRUE) + } + cv.sd[j] <- sqrt(var(sds))/mean(sds) + } + o = (1:length(s0.perc))[cv.sd == min(cv.sd)] + # we don;t allow taking s0.hat to be 0th percentile when + # min sd is 0 + s0.hat = quantile(sd[sd != 0], s0.perc[o]) + return(list(s0.perc = s0.perc, cv.sd = cv.sd, s0.hat = s0.hat)) +} + +permute.rows <- function(x) { + dd <- dim(x) + n <- dd[1] + p <- dd[2] + mm <- runif(length(x)) + rep(seq(n) * 10, rep(p, n)) + matrix(t(x)[order(mm)], n, p, byrow = TRUE) +} + +foldchange.paired = function(x, y, logged2) { + # if(logged2){x=2^x} + nc <- ncol(x)/2 + o <- 1:nc + o1 <- rep(0, ncol(x)/2) + o2 <- o1 + for (j in 1:nc) { + o1[j] <- (1:ncol(x))[y == -o[j]] + } + for (j in 1:nc) { + o2[j] <- (1:ncol(x))[y == o[j]] + } + if (!logged2) { + d <- x[, o2, drop = F]/x[, o1, drop = F] + } + if (logged2) { + d <- x[, o2, drop = F] - x[, o1, drop = F] + } + if (!logged2) { + fc <- rowMeans(d) + } + if (logged2) { + fc <- 2^rowMeans(d) + } + return(fc) +} foldchange.seq.twoclass.unpaired <- function(x, y, depth) { x.norm <- scale(x, center = F, scale = depth) + 1e-08 @@ -780,3 +1156,227 @@ foldchange.seq.twoclass.unpaired <- function(x, y, depth) 1]) return(fc) } +integer.base.b <- function(x, b = 2) { + xi <- as.integer(x) + if (xi == 0) { + return(0) + } + if (any(is.na(xi) | ((x - xi) != 0))) + print(list(ERROR = "x not integer", x = x)) + N <- length(x) + xMax <- max(x) + ndigits <- (floor(logb(xMax, base = 2)) + 1) + Base.b <- array(NA, dim = c(N, ndigits)) + for (i in 1:ndigits) { + #i <- 1 + Base.b[, ndigits - i + 1] <- (x%%b) + x <- (x%/%b) + } + if (N == 1) + Base.b[1, ] + else Base.b +} +varr <- function(x, meanx = NULL) { + n <- ncol(x) + p <- nrow(x) + Y <- matrix(1, nrow = n, ncol = 1) + if (is.null(meanx)) { + meanx <- rowMeans(x) + } + ans <- rep(1, p) + xdif <- x - meanx %*% t(Y) + ans <- (xdif^2) %*% rep(1/(n - 1), n) + ans <- drop(ans) + return(ans) +} +quantitative.func <- function(x, y, s0 = 0) { + # regression of x on y + my = mean(y) + yy <- y - my + temp <- x %*% yy + mx = rowMeans(x) + syy = sum(yy^2) + scor <- temp/syy + b0hat <- mx - scor * my + ym = matrix(y, nrow = nrow(x), ncol = ncol(x), byrow = T) + xhat <- matrix(b0hat, nrow = nrow(x), ncol = ncol(x)) + ym * + matrix(scor, nrow = nrow(x), ncol = ncol(x)) + sigma <- sqrt(rowSums((x - xhat)^2)/(ncol(xhat) - 2)) + sd <- sigma/sqrt(syy) + tt <- scor/(sd + s0) + return(list(tt = tt, numer = scor, sd = sd)) +} +timearea.func <- function(x, y, s0 = 0) { + n <- ncol(x) + xx <- 0.5 * (x[, 2:n] + x[, 1:(n - 1)]) * matrix(diff(y), + nrow = nrow(x), ncol = n - 1, byrow = T) + numer <- rowMeans(xx) + sd <- sqrt(varr(xx, meanx = numer)/n) + tt <- numer/sqrt(sd + s0) + return(list(tt = tt, numer = numer, sd = sd)) +} +cox.seq.func <- function(xresamp, y, censoring.status) { + # get the dimensions + ng <- dim(xresamp)[1] + ns <- dim(xresamp)[2] + # prepare for the calculation + # find the index matrix + Dn <- sum(censoring.status == 1) + Dset <- c(1:ns)[censoring.status == 1] # the set of died + ind <- matrix(0, ns, Dn) + # get the matrix + for (i in 1:Dn) { + ind[y >= y[Dset[i]] - 1e-08, i] <- 1/sum(y >= y[Dset[i]] - + 1e-08) + } + ind.sums <- rowSums(ind) + # calculate the score statistic + tt <- apply(xresamp, 3, function(x, cen.ind, ind.para, ind.sums.para) { + dev1 <- x %*% cen.ind + x.ind <- x %*% ind.para + dev2 <- (x * x) %*% ind.sums.para - rowSums(x.ind * x.ind) + dev1/(sqrt(dev2) + 1e-08) + }, (censoring.status - ind.sums), ind, ind.sums) + tt <- rowMeans(tt) + return(list(tt = tt, numer = tt, sd = rep(1, length(tt)))) +} +compute.block.perms = function(y, blocky, nperms) { + # y are the data (eg class label 1 vs 2; or -1,1, -2,2 for + # paired data) + # blocky are the block labels (abs(y) for paired daatr) + ny = length(y) + nblocks = length(unique(blocky)) + tab = table(blocky) + total.nperms = prod(factorial(tab)) + # block.perms is a list of all possible permutations + block.perms = vector("list", nblocks) + # first enumerate all perms, when possible + if (total.nperms <= nperms) { + all.perms.flag = 1 + nperms.act = total.nperms + for (i in 1:nblocks) { + block.perms[[i]] = permute(y[blocky == i]) + } + kk = 0:(factorial(max(tab))^nblocks - 1) + #the rows of the matrix outerm runs through the 'outer + # product' + # first we assume that all blocks have max(tab) members; + # then we remove rows of outerm that + # are illegal (ie when a block has fewer members) + outerm = matrix(0, nrow = length(kk), ncol = nblocks) + for (i in 1:length(kk)) { + kkkk = integer.base.b(kk[i], b = factorial(max(tab))) + if (length(kkkk) > nblocks) { + kkkk = kkkk[(length(kkkk) - nblocks + 1):length(kkkk)] + } + outerm[i, (nblocks - length(kkkk) + 1):nblocks] = kkkk + } + outerm = outerm + 1 + # now remove rows that are illegal perms + ind = rep(TRUE, nrow(outerm)) + for (j in 1:ncol(outerm)) { + ind = ind & outerm[, j] <= factorial(tab[j]) + } + outerm = outerm[ind, , drop = F] + # finally, construct permutation matrix from outer product + permsy = matrix(NA, nrow = total.nperms, ncol = ny) + for (i in 1:total.nperms) { + junk = NULL + for (j in 1:nblocks) { + junk = c(junk, block.perms[[j]][outerm[i, j], + ]) + } + permsy[i, ] = junk + } + } + # next handle case when there are too many perms to enumerate + if (total.nperms > nperms) { + all.perms.flag = 0 + nperms.act = nperms + permsy = NULL + block.perms = vector("list", nblocks) + for (j in 1:nblocks) { + block.perms[[j]] = sample.perms(y[blocky == j], nperms = nperms) + } + for (j in 1:nblocks) { + permsy = cbind(permsy, block.perms[[j]]) + } + } + return(list(permsy = permsy, all.perms.flag = all.perms.flag, + nperms.act = nperms.act)) +} +sample.perms <- function(elem, nperms) { + # randomly generates nperms of the vector elem + res = permute.rows(matrix(elem, nrow = nperms, ncol = length(elem), + byrow = T)) + return(res) +} +mysvd <- function(x, n.components = NULL) { + # finds PCs of matrix x + p <- nrow(x) + n <- ncol(x) + # center the observations (rows) + feature.means <- rowMeans(x) + x <- t(scale(t(x), center = feature.means, scale = F)) + if (is.null(n.components)) { + n.components = min(n, p) + } + if (p > n) { + a <- eigen(t(x) %*% x) + v <- a$vec[, 1:n.components, drop = FALSE] + d <- sqrt(a$val[1:n.components, drop = FALSE]) + u <- scale(x %*% v, center = FALSE, scale = d) + return(list(u = u, d = d, v = v)) + } + else { + junk <- svd(x, LINPACK = TRUE) + nc = min(ncol(junk$u), n.components) + return(list(u = junk$u[, 1:nc], d = junk$d[1:nc], v = junk$v[, + 1:nc])) + } +} +quantitative.seq.func <- function(xresamp, y) { + tt <- rep(0, dim(xresamp)[1]) + for (i in 1:dim(xresamp)[3]) { + y.ranked <- rank(y, ties.method = "random") - (dim(xresamp)[2] + + 1)/2 + tt <- tt + (xresamp[, , i] - (dim(xresamp)[2] + 1)/2) %*% + y.ranked + } + ns <- dim(xresamp)[2] + tt <- tt/(dim(xresamp)[3] * (ns^3 - ns)/12) + return(list(tt = as.vector(tt), numer = as.vector(tt), sd = rep(1, + length(tt)))) +} +multiclass.seq.func <- function(xresamp, y) +{ + # number of classes and number of samples in each class + K <- max(y) + n.each <- rep(0, K) + for (k in 1 : K) + { + n.each[k] <- sum(y == k) + } + # the statistic + tt <- temp <- rep(0, dim(xresamp)[1]) + stand.contrasts <- matrix(0, dim(xresamp)[1], K) + + for (i in 1 : dim(xresamp)[3]) + { + for (k in 1 : K) + { + temp <- rowSums(xresamp[, y == k, i]) + tt <- tt + temp ^2 / n.each[k] + stand.contrasts[, k] <- stand.contrasts[, k] + temp + } + } + # finalize + nresamp <- dim(xresamp)[3] + ns <- dim(xresamp)[2] + tt <- tt / nresamp * 12 / ns / (ns + 1) - 3 * (ns + 1) + stand.contrasts <- stand.contrasts / nresamp + stand.contrasts <- scale(stand.contrasts, center=n.each * (ns + 1) / 2, + scale=sqrt(n.each * (ns - n.each) * (ns + 1) / 12)) + return(list(tt = tt, numer = tt, sd = rep(1, length(tt)), + stand.contrasts = stand.contrasts)) +} diff --git a/man/check.format.Rd b/man/check.format.Rd index dcee24c..452ca0e 100644 --- a/man/check.format.Rd +++ b/man/check.format.Rd @@ -1,11 +1,18 @@ % Generated by roxygen2: do not edit by hand -% Please edit documentation in R/samr.R +% Please edit documentation in R/samr-adapted.R \name{check.format} \alias{check.format} \title{Check format} \usage{ check.format(y, resp.type, censoring.status = NULL) } +\arguments{ +\item{y}{y} + +\item{resp.type}{resp type} + +\item{censoring.status}{censoring status} +} \description{ Check format } diff --git a/man/foldchange.seq.twoclass.unpaired.Rd b/man/foldchange.seq.twoclass.unpaired.Rd index b95217e..bcc8668 100644 --- a/man/foldchange.seq.twoclass.unpaired.Rd +++ b/man/foldchange.seq.twoclass.unpaired.Rd @@ -1,11 +1,18 @@ % Generated by roxygen2: do not edit by hand -% Please edit documentation in R/samr.R +% Please edit documentation in R/samr-adapted.R \name{foldchange.seq.twoclass.unpaired} \alias{foldchange.seq.twoclass.unpaired} \title{Foldchange of twoclass unpaired sequencing data} \usage{ foldchange.seq.twoclass.unpaired(x, y, depth) } +\arguments{ +\item{x}{x} + +\item{y}{y} + +\item{depth}{depth} +} \description{ Foldchange of twoclass unpaired sequencing data } diff --git a/man/rankcols.Rd b/man/rankcols.Rd index 09d9e8b..b3abb74 100644 --- a/man/rankcols.Rd +++ b/man/rankcols.Rd @@ -1,11 +1,14 @@ % Generated by roxygen2: do not edit by hand -% Please edit documentation in R/samr.R +% Please edit documentation in R/samr-adapted.R \name{rankcols} \alias{rankcols} \title{Rank columns} \usage{ rankcols(x) } +\arguments{ +\item{x}{x} +} \description{ Ranks the elements within each col of the matrix x and returns these ranks in a matrix diff --git a/man/resa.Rd b/man/resa.Rd index 74b9d5f..1a6dee8 100644 --- a/man/resa.Rd +++ b/man/resa.Rd @@ -1,5 +1,5 @@ % Generated by roxygen2: do not edit by hand -% Please edit documentation in R/samr.R +% Please edit documentation in R/samr-adapted.R \name{resa} \alias{resa} \title{Resampling} diff --git a/man/sammy.Rd b/man/sammy.Rd index a3d45e7..db447ff 100644 --- a/man/sammy.Rd +++ b/man/sammy.Rd @@ -1,5 +1,5 @@ % Generated by roxygen2: do not edit by hand -% Please edit documentation in R/samr.R +% Please edit documentation in R/samr-adapted.R \name{sammy} \alias{sammy} \title{Significance analysis of microarrays} @@ -27,6 +27,43 @@ sammy( xl.prevfit = NULL ) } +\arguments{ +\item{data}{Data object with components x- p by n matrix of features, one observation per column (missing values allowed); y- n-vector of outcome measurements; censoring.status- n-vector of censoring censoring.status (1= died or event occurred, 0=survived, or event was censored), needed for a censored survival outcome} + +\item{resp.type}{Problem type: "Quantitative" for a continuous parameter (Available for both array and sequencing data); "Two class unpaired" (for both array and sequencing data); "Survival" for censored survival outcome (for both array and sequencingdata); "Multiclass": more than 2 groups (for both array and sequencing data); "One class" for a single group (only for array data); "Two class paired" for two classes with paired observations (for both array and sequencing data); "Two class unpaired timecourse" (only for array data), "One class time course" (only for array data), "Two class.paired timecourse" (only for array data), or "Pattern discovery" (only for array data)} + +\item{assay.type}{Assay type: "array" for microarray data, "seq" for counts from sequencing} + +\item{s0}{Exchangeability factor for denominator of test statistic; Default is automatic choice. Only used for array data.} + +\item{s0.perc}{Percentile of standard deviation values to use for s0; default is automatic choice; -1 means s0=0 (different from s0.perc=0, meaning s0=zeroeth percentile of standard deviation values= min of sd values. Only used for array data.} + +\item{nperms}{Number of permutations used to estimate false discovery rates} + +\item{center.arrays}{Should the data for each sample (array) be median centered at the outset? Default =FALSE. Only used for array data.} + +\item{testStatistic}{Test statistic to use in two class unpaired case.Either "standard" (t-statistic) or ,"wilcoxon" (Two-sample wilcoxon or Mann-Whitney test). Only used for array data.} + +\item{time.summary.type}{Summary measure for each time course: "slope", or"signed.area"). Only used for array data.} + +\item{regression.method}{Regression method for quantitative case: "standard",(linear least squares) or "ranks" (linear least squares on ranked data). Only used for array data.} + +\item{return.x}{Should the matrix of feature values be returned? Only useful for time course data, where x contains summaries of the features over time. Otherwise x is the same as the input data data\$x} + +\item{knn.neighbors}{Number of nearest neighbors to use for imputation of missing features values. Only used for array data.} + +\item{random.seed}{Optional initial seed for random number generator (integer)} + +\item{nresamp}{For assay.type="seq", number of resamples used to construct test statistic. Default 20. Only used for sequencing data.} + +\item{nresamp.perm}{For assay.type="seq", number of resamples used to construct test statistic for permutations. Default is equal to nresamp and it must be at most nresamp. Only used for sequencing data.} + +\item{xl.mode}{Used by Excel interface} + +\item{xl.time}{Used by Excel interface} + +\item{xl.prevfit}{Used by Excel interface} +} \description{ This function is an adaptation of `samr::samr` } diff --git a/man/samr.estimate.depth.Rd b/man/samr.estimate.depth.Rd index f91d5e5..75ab46e 100644 --- a/man/samr.estimate.depth.Rd +++ b/man/samr.estimate.depth.Rd @@ -1,5 +1,5 @@ % Generated by roxygen2: do not edit by hand -% Please edit documentation in R/samr.R +% Please edit documentation in R/samr-adapted.R \name{samr.estimate.depth} \alias{samr.estimate.depth} \title{Estimate sequencing depths} diff --git a/man/wilcoxon.unpaired.seq.func.Rd b/man/wilcoxon.unpaired.seq.func.Rd index 1925ecc..aadb6ee 100644 --- a/man/wilcoxon.unpaired.seq.func.Rd +++ b/man/wilcoxon.unpaired.seq.func.Rd @@ -1,5 +1,5 @@ % Generated by roxygen2: do not edit by hand -% Please edit documentation in R/samr.R +% Please edit documentation in R/samr-adapted.R \name{wilcoxon.unpaired.seq.func} \alias{wilcoxon.unpaired.seq.func} \title{Twoclass Wilcoxon statistics} From ded4d1a1f36fe191ea1b07fea3d57e7911073f04 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Tue, 26 May 2020 12:01:11 +0200 Subject: [PATCH 06/99] Improved unit tests --- tests/testthat/test.DEGfreeze.R | 46 +++++++------------------------ tests/testthat/test.DIscBIO.IMP.R | 8 +++--- 2 files changed, 14 insertions(+), 40 deletions(-) diff --git a/tests/testthat/test.DEGfreeze.R b/tests/testthat/test.DEGfreeze.R index 96c396e..7d0c1ce 100644 --- a/tests/testthat/test.DEGfreeze.R +++ b/tests/testthat/test.DEGfreeze.R @@ -1,61 +1,35 @@ -# library(devtools)#TEMP -# library(testthat)#TEMP -# source("Aux/samr.R")#TEMP -# source("Aux/resample.R")#TEMP -# source("Aux/rankcol.R")#TEMP -# source("Aux/samr/R/samr.morefuns.R")#TEMP -# source("Aux/samr/R/samr.R")#TEMP -# library(DIscBIO)#TEMP context("Stress-testing DEG analyses") -# ============================================================================== -# Loading dataset -# ============================================================================== -data(pan_indrop_matrix_8000cells_18556genes) - # ============================================================================== # Determining contants # ============================================================================== n_genes <- 1e3 n_clust <- 3 - +manual_test <- FALSE # ============================================================================== # Defining datasets to be used # ============================================================================== -message(Sys.time(), " - Subsetting") -sc <- DISCBIO(pan_indrop_matrix_8000cells_18556genes[, seq_len(n_genes)]) +if (manual_test) { + data(pan_indrop_matrix_8000cells_18556genes) + sc <- DISCBIO(pan_indrop_matrix_8000cells_18556genes[, seq_len(n_genes)]) +} else { + sc <- DISCBIO(valuesG1msReduced) + sc <- NoiseFiltering(sc, plot=FALSE, export=FALSE, quiet=TRUE) +} # ============================================================================== # Performing operations # ============================================================================== -# MIinExp <- mean(rowMeans(dataset, na.rm=TRUE)) -# MinNumber <- round(length(dataset[1, ]) / 10) # 2b xpressed in > 10% cells. -# sc <- Normalizedata( -# sc, mintotal=1000, minexpr=MIinExp, minnumber=MinNumber, maxexpr=Inf, -# downsample=FALSE, dsn=1, rseed=17000 -# ) -message("Filtering") -sc <- Normalizedata(sc, mintotal=1000, maxexpr=Inf, downsample=FALSE, dsn=1) +sc <- Normalizedata(sc) sc <- FinalPreprocessing(sc, GeneFlitering="ExpF", export=FALSE, quiet=TRUE) -sc <- Clustexp(sc, clustnr=n_clust, quiet=TRUE, bootnr = 2, B.gap = 2) # ------------------------------------------------------------------------------ # DEG Analysis # ------------------------------------------------------------------------------ -message("Analysing DEG") +sc <- Clustexp(sc, clustnr=n_clust, quiet=TRUE, bootnr = 2, B.gap = 2) cdiff <- DEGanalysis( sc, Clustering="K-means", K=n_clust, fdr=0.10, name="all_clusters", export=FALSE, quiet=TRUE, plot=FALSE, nresamp=2, nperms=2 ) -# sc <- Exprmclust(sc, K=K, reduce=TRUE, quiet=TRUE) -# sc <- MB_Order(sc, quiet=TRUE, export=FALSE) -# MBcdiffBinomial <- MBClustDiffGenes( -# sc, K=K, pValue=0.05, fdr=0.15, export=FALSE, quiet=FALSE -# ) -# sc <- Clustexp(sc, cln=K, quiet=TRUE) -# MBcdiff <- DEGanalysis( -# sc, Clustering="MB", K=K, fdr=0.05, name="all_clusters", export = FALSE, -# quiet=TRUE -# ) test_that("DEG analysis doesn't freeze", { expect_output(str(cdiff), "List of 2") }) \ No newline at end of file diff --git a/tests/testthat/test.DIscBIO.IMP.R b/tests/testthat/test.DIscBIO.IMP.R index 8ade01f..95347ed 100644 --- a/tests/testthat/test.DIscBIO.IMP.R +++ b/tests/testthat/test.DIscBIO.IMP.R @@ -71,14 +71,14 @@ test_that("Cluster plots output is as expexted", { context("Outliers") Outliers <- FindOutliersKM( - sc, K=2, outminc=5, outlg=2, probthr=.5*1e-3, thr=2**-(1:40), + sc, K=2, outminc=5, outlg=2, probthr=.5*1e-3, thr=2 ^ (-1:-40), outdistquant=.75, plot = FALSE, quiet = TRUE ) # Adjusting outliers outlg <- round(length(sc@fdata[, 1]) / 200) # The cell will be considered as an outlier if it has a minimum of 0.5% of the number of filtered genes as outlier genes. Outliers2 <- FindOutliersKM( - sc, K=2, outminc=5, outlg=outlg, probthr=.5*1e-3, thr=2**-(1:40), + sc, K=2, outminc=5, outlg=outlg, probthr=.5*1e-3, thr=2 ^ (-1:-40), outdistquant=.75, plot = FALSE, quiet = TRUE ) Order <- KmeanOrder(sc, quiet = TRUE, export = FALSE) @@ -185,12 +185,12 @@ context("MB outliers") sc <- Clustexp(sc, cln=2, quiet=TRUE) Outliers <- FindOutliersMB( - sc, K=2, outminc=5, outlg=2, probthr=.5*1e-3, thr=2**-(1:40), + sc, K=2, outminc=5, outlg=2, probthr=.5*1e-3, thr=2 ^ (-1:-40), outdistquant=.75, plot = FALSE, quiet = TRUE ) outlg <- round(length(sc@fdata[, 1]) / 200) # The cell will be considered as an outlier if it has a minimum of 0.5% of the number of filtered genes as outlier genes. Outliers2 <- FindOutliersMB( - sc, K=2, outminc=5, outlg=outlg, probthr=.5*1e-3, thr=2**-(1:40), + sc, K=2, outminc=5, outlg=outlg, probthr=.5*1e-3, thr=2 ^ (-1:-40), outdistquant=.75, plot = FALSE, quiet = TRUE ) From 9b7256a322c864fa21d0da41f627df9831a43646 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Tue, 16 Jun 2020 10:59:40 +0200 Subject: [PATCH 07/99] Improved formatting --- DESCRIPTION | 160 +++++++++++++++++++++++++++------------------------- 1 file changed, 84 insertions(+), 76 deletions(-) diff --git a/DESCRIPTION b/DESCRIPTION index 8937ae7..630b764 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -3,30 +3,38 @@ Date: 2020-05-04 Title: A User-Friendly Pipeline for Biomarker Discovery in Single-Cell Transcriptomics Version: 0.99.8 Authors@R: - c( - person(given = "Salim", - family = "Ghannoum", - role = c("aut", "cph"), - email = "salim.ghannoum@medisin.uio.no"), - person(given = "Alvaro", - family = "Köhn-Luque", - role = c("aut", "ths"), - email = "alvaro.kohn-luque@medisin.uio.no"), - person(given = "Waldir", - family = "Leoncio", - role = c("cre", "aut"), - email = "w.l.netto@medisin.uio.no"), - person(given = "Damiano", - family = "Fantini", - role = c("ctb")) - ) + c( + person( + given = "Salim", + family = "Ghannoum", + role = c("aut", "cph"), + email = "salim.ghannoum@medisin.uio.no" + ), + person( + given = "Alvaro", + family = "Köhn-Luque", + role = c("aut", "ths"), + email = "alvaro.kohn-luque@medisin.uio.no" + ), + person( + given = "Waldir", + family = "Leoncio", + role = c("cre", "aut"), + email = "w.l.netto@medisin.uio.no" + ), + person( + given = "Damiano", + family = "Fantini", + role = c("ctb") + ) + ) Description: An open, multi-algorithmic pipeline for easy, fast and efficient - analysis of cellular sub-populations and the molecular signatures that - characterize them. The pipeline consists of four successive steps: data - pre-processing, cellular clustering with pseudo-temporal ordering, defining - differential expressed genes and biomarker identification. This package - implements extensions of the work published by Ghannoum et. al. - . + analysis of cellular sub-populations and the molecular signatures that + characterize them. The pipeline consists of four successive steps: data + pre-processing, cellular clustering with pseudo-temporal ordering, defining + differential expressed genes and biomarker identification. This package + implements extensions of the work published by Ghannoum et. al. + . biocViews: SingleCell, Transcriptomics, Clustering, DecisionTree License: MIT + file LICENSE Encoding: UTF-8 @@ -35,63 +43,63 @@ Imports: methods, TSCAN, boot, httr, mclust, statmod, biomaRt, samr, igraph, readr, RColorBrewer, ggplot2, rpart, fpc, cluster, rpart.plot, amap, dplyr, tsne, AnnotationDbi, org.Hs.eg.db, calibrate, graphics, stats, utils, impute Depends: - R (>= 4.0), SingleCellExperiment + R (>= 4.0), SingleCellExperiment Suggests: - knitr, - rmarkdown, + knitr, + rmarkdown, testthat, - enrichR, - Seurat + enrichR, + Seurat VignetteBuilder: knitr LazyData: true RoxygenNote: 7.1.0 BugReports: https://github.com/ocbe-uio/DIscBIO/issues Collate: - 'DIscBIO-classes.R' - 'DIscBIO-generic-ClassVectoringDT.R' - 'DIscBIO-generic-Clustexp.R' - 'DIscBIO-generic-DEGanalysis.R' - 'DIscBIO-generic-DEGanalysis2clust.R' - 'DIscBIO-generic-Exprmclust.R' - 'DIscBIO-generic-FinalPreprocessing.R' - 'DIscBIO-generic-FindOutliersKM.R' - 'DIscBIO-generic-FindOutliersMB.R' - 'DIscBIO-generic-KMClustDiffGenes.R' - 'DIscBIO-generic-KMclustheatmap.R' - 'DIscBIO-generic-KmeanOrder.R' - 'DIscBIO-generic-MBClustDiffGenes.R' - 'DIscBIO-generic-MBclustheatmap.R' - 'DIscBIO-generic-NoiseFiltering.R' - 'DIscBIO-generic-Normalizedata.R' - 'DIscBIO-generic-PCAplotSymbols.R' - 'DIscBIO-generic-PlotmclustMB.R' - 'DIscBIO-generic-comptSNE.R' - 'DIscBIO-generic-comptsneMB.R' - 'DIscBIO-generic-plotExptSNE.R' - 'DIscBIO-generic-plotGap.R' - 'DIscBIO-generic-plotKmeansLabelstSNE.R' - 'DIscBIO-generic-plotMBLabelstSNE.R' - 'DIscBIO-generic-plotOrderKMtsne.R' - 'DIscBIO-generic-plotOrderMBtsne.R' - 'DIscBIO-generic-plotSilhouette.R' - 'DIscBIO-generic-plotSymbolstSNE.R' - 'DIscBIO-generic-plotexptsneMB.R' - 'DIscBIO-generic-plotsilhouetteMB.R' - 'DIscBIO-generic-plottSNE.R' - 'DIscBIO-generic-plottsneMB.R' - 'J48DT.R' - 'J48DTeval.R' - 'Jaccard.R' - 'MB_Order.R' - 'NetAnalysis.R' - 'Networking.R' - 'PPI.R' - 'PlotMBexpPCA.R' - 'PlotMBorderPCA.R' - 'RpartDT.R' - 'RpartEVAL.R' - 'VolcanoPlot.R' - 'customConverters.R' - 'datasets.R' - 'retrieveBiomart.R' - 'samr-adapted.R' + 'DIscBIO-classes.R' + 'DIscBIO-generic-ClassVectoringDT.R' + 'DIscBIO-generic-Clustexp.R' + 'DIscBIO-generic-DEGanalysis.R' + 'DIscBIO-generic-DEGanalysis2clust.R' + 'DIscBIO-generic-Exprmclust.R' + 'DIscBIO-generic-FinalPreprocessing.R' + 'DIscBIO-generic-FindOutliersKM.R' + 'DIscBIO-generic-FindOutliersMB.R' + 'DIscBIO-generic-KMClustDiffGenes.R' + 'DIscBIO-generic-KMclustheatmap.R' + 'DIscBIO-generic-KmeanOrder.R' + 'DIscBIO-generic-MBClustDiffGenes.R' + 'DIscBIO-generic-MBclustheatmap.R' + 'DIscBIO-generic-NoiseFiltering.R' + 'DIscBIO-generic-Normalizedata.R' + 'DIscBIO-generic-PCAplotSymbols.R' + 'DIscBIO-generic-PlotmclustMB.R' + 'DIscBIO-generic-comptSNE.R' + 'DIscBIO-generic-comptsneMB.R' + 'DIscBIO-generic-plotExptSNE.R' + 'DIscBIO-generic-plotGap.R' + 'DIscBIO-generic-plotKmeansLabelstSNE.R' + 'DIscBIO-generic-plotMBLabelstSNE.R' + 'DIscBIO-generic-plotOrderKMtsne.R' + 'DIscBIO-generic-plotOrderMBtsne.R' + 'DIscBIO-generic-plotSilhouette.R' + 'DIscBIO-generic-plotSymbolstSNE.R' + 'DIscBIO-generic-plotexptsneMB.R' + 'DIscBIO-generic-plotsilhouetteMB.R' + 'DIscBIO-generic-plottSNE.R' + 'DIscBIO-generic-plottsneMB.R' + 'J48DT.R' + 'J48DTeval.R' + 'Jaccard.R' + 'MB_Order.R' + 'NetAnalysis.R' + 'Networking.R' + 'PPI.R' + 'PlotMBexpPCA.R' + 'PlotMBorderPCA.R' + 'RpartDT.R' + 'RpartEVAL.R' + 'VolcanoPlot.R' + 'customConverters.R' + 'datasets.R' + 'retrieveBiomart.R' + 'samr-adapted.R' From 69238cd7897c22a23a1210b14456c9aa663eb414 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Tue, 16 Jun 2020 15:07:00 +0200 Subject: [PATCH 08/99] Minor modifications to DESCRIPTION for CRAN submission --- DESCRIPTION | 8 ++++---- 1 file changed, 4 insertions(+), 4 deletions(-) diff --git a/DESCRIPTION b/DESCRIPTION index 630b764..68d1310 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -1,7 +1,7 @@ Package: DIscBIO -Date: 2020-05-04 +Date: 2020-06-16 Title: A User-Friendly Pipeline for Biomarker Discovery in Single-Cell Transcriptomics -Version: 0.99.8 +Version: 1.0.0 Authors@R: c( person( @@ -35,7 +35,6 @@ Description: An open, multi-algorithmic pipeline for easy, fast and efficient differential expressed genes and biomarker identification. This package implements extensions of the work published by Ghannoum et. al. . -biocViews: SingleCell, Transcriptomics, Clustering, DecisionTree License: MIT + file LICENSE Encoding: UTF-8 Imports: methods, TSCAN, boot, httr, mclust, statmod, biomaRt, samr, igraph, @@ -49,7 +48,8 @@ Suggests: rmarkdown, testthat, enrichR, - Seurat + Seurat, + BiocManager VignetteBuilder: knitr LazyData: true RoxygenNote: 7.1.0 From 629f4f99e15952e6b7245dcec61623899de5bcf4 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 22 Jun 2020 11:32:33 +0200 Subject: [PATCH 09/99] Matched LICENSE to CRAN template --- LICENSE | 23 ++--------------------- 1 file changed, 2 insertions(+), 21 deletions(-) diff --git a/LICENSE b/LICENSE index 80db17c..eaa36da 100644 --- a/LICENSE +++ b/LICENSE @@ -1,21 +1,2 @@ -MIT License - -Copyright (c) 2019 SystemsBiologist - -Permission is hereby granted, free of charge, to any person obtaining a copy -of this software and associated documentation files (the "Software"), to deal -in the Software without restriction, including without limitation the rights -to use, copy, modify, merge, publish, distribute, sublicense, and/or sell -copies of the Software, and to permit persons to whom the Software is -furnished to do so, subject to the following conditions: - -The above copyright notice and this permission notice shall be included in all -copies or substantial portions of the Software. - -THE SOFTWARE IS PROVIDED "AS IS", WITHOUT WARRANTY OF ANY KIND, EXPRESS OR -IMPLIED, INCLUDING BUT NOT LIMITED TO THE WARRANTIES OF MERCHANTABILITY, -FITNESS FOR A PARTICULAR PURPOSE AND NONINFRINGEMENT. IN NO EVENT SHALL THE -AUTHORS OR COPYRIGHT HOLDERS BE LIABLE FOR ANY CLAIM, DAMAGES OR OTHER -LIABILITY, WHETHER IN AN ACTION OF CONTRACT, TORT OR OTHERWISE, ARISING FROM, -OUT OF OR IN CONNECTION WITH THE SOFTWARE OR THE USE OR OTHER DEALINGS IN THE -SOFTWARE. \ No newline at end of file +YEAR: 2019 +COPYRIGHT HOLDER: SystemsBiologist \ No newline at end of file From 443ec3fac43107ba1b00942dfa08f76ed1e8a52d Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 22 Jun 2020 11:33:01 +0200 Subject: [PATCH 10/99] Changed Weka test units --- tests/testthat/test.DIscBIO.IMP.R | 8 ++++++-- 1 file changed, 6 insertions(+), 2 deletions(-) diff --git a/tests/testthat/test.DIscBIO.IMP.R b/tests/testthat/test.DIscBIO.IMP.R index 95347ed..2b128e7 100644 --- a/tests/testthat/test.DIscBIO.IMP.R +++ b/tests/testthat/test.DIscBIO.IMP.R @@ -149,7 +149,9 @@ rpartEVAL <- RpartEVAL( test_that("Decision tree elements are defined", { expect_output(str(DATAforDT), "3 obs. of 30 variables") expect_s3_class(j48dt, "J48") - expect_s3_class(summary(j48dt), "Weka_classifier_evaluation") + expect_identical( + attributes(j48dt)$class, c("J48", "Weka_tree", "Weka_classifier") + ) expect_identical(j48dt_eval, c(TP = 14, FN = 4, FP = 3, TN = 9)) expect_s3_class(rpartDT, "rpart") expect_identical(rpartEVAL, c(TP = 16, FN = 2, FP = 4, TN = 8)) @@ -270,7 +272,9 @@ rpartEVAL <- RpartEVAL( test_that("Decision tree elements are defined", { expect_output(str(DATAforDT), "29 obs. of 30 variables") # used to be 31 expect_s3_class(j48dt, "J48") - expect_s3_class(summary(j48dt), "Weka_classifier_evaluation") + expect_identical( + attributes(j48dt)$class, c("J48", "Weka_tree", "Weka_classifier") + ) expect_identical(j48dt_eval, c(TP = 21, FN = 2, FP = 4, TN = 3)) expect_s3_class(rpartDT, "rpart") expect_identical(rpartEVAL, c(TP = 20, FN = 3, FP = 4, TN = 3)) From fdc9415c549bb5eb9d4f189799bb141bff73a92d Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 22 Jun 2020 11:49:19 +0200 Subject: [PATCH 11/99] Reduced example duration --- R/DIscBIO-generic-DEGanalysis.R | 3 ++- R/J48DT.R | 3 ++- 2 files changed, 4 insertions(+), 2 deletions(-) diff --git a/R/DIscBIO-generic-DEGanalysis.R b/R/DIscBIO-generic-DEGanalysis.R index c7803c1..4364e25 100644 --- a/R/DIscBIO-generic-DEGanalysis.R +++ b/R/DIscBIO-generic-DEGanalysis.R @@ -30,7 +30,8 @@ #' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) #' sc <- comptSNE(sc, max_iter=100) #' DEGanalysis( -#' sc, Clustering="K-means", K=3, fdr=0.1, name="Name", export = FALSE +#' sc, Clustering="K-means", K=3, fdr=0.1, name="Name", export = FALSE, +#' nresamp=5, nperms=20 #' ) setGeneric( name = "DEGanalysis", diff --git a/R/J48DT.R b/R/J48DT.R index 3618d07..c62015c 100644 --- a/R/J48DT.R +++ b/R/J48DT.R @@ -24,7 +24,8 @@ #' sc <- comptSNE(sc, max_iter=100) #' cdiff <- DEGanalysis2clust( #' sc, Clustering="K-means", K=3, fdr=.2, name="Name", First="CL1", -#' Second="CL2", export=FALSE +#' Second="CL2", export=FALSE, +#' nresamp=5, nperms=20 #' ) #' sigDEG <- cdiff[[1]] #' DATAforDT <- ClassVectoringDT( From d23b6230f4fcd3eadcda3dc90b2cc58e6e1e805c Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 29 Jun 2020 10:58:30 +0200 Subject: [PATCH 12/99] Changed dontrun to donttest --- R/DIscBIO-generic-MBClustDiffGenes.R | 2 +- R/DIscBIO-generic-MBclustheatmap.R | 2 +- R/DIscBIO-generic-plotexptsneMB.R | 2 +- R/MB_Order.R | 2 +- R/NetAnalysis.R | 2 +- R/Networking.R | 2 +- R/PPI.R | 2 +- R/VolcanoPlot.R | 2 +- man/DEGanalysis.Rd | 3 ++- man/J48DT.Rd | 3 ++- man/MBClustDiffGenes.Rd | 2 +- man/MB_Order.Rd | 2 +- man/MBclustheatmap.Rd | 2 +- man/NetAnalysis.Rd | 2 +- man/Networking.Rd | 2 +- man/PPI.Rd | 2 +- man/VolcanoPlot.Rd | 2 +- man/plotexptsneMB.Rd | 2 +- 18 files changed, 20 insertions(+), 18 deletions(-) diff --git a/R/DIscBIO-generic-MBClustDiffGenes.R b/R/DIscBIO-generic-MBClustDiffGenes.R index 2c7f1f0..9c06e05 100644 --- a/R/DIscBIO-generic-MBClustDiffGenes.R +++ b/R/DIscBIO-generic-MBClustDiffGenes.R @@ -13,7 +13,7 @@ #' @export #' @return A list containing two tables. #' @examples -#' \dontrun{ +#' \donttest{ #' sc <- DISCBIO(valuesG1msReduced) #' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) #' sc <- Normalizedata( diff --git a/R/DIscBIO-generic-MBclustheatmap.R b/R/DIscBIO-generic-MBclustheatmap.R index b511403..eee63c0 100644 --- a/R/DIscBIO-generic-MBclustheatmap.R +++ b/R/DIscBIO-generic-MBclustheatmap.R @@ -15,7 +15,7 @@ #' @return Unless otherwise specified, a heatmap and a vector of the underlying #' cluster order. #' @examples -#' \dontrun{ +#' \donttest{ #' sc <- DISCBIO(valuesG1msReduced) #' sc <- NoiseFiltering(sc, export=FALSE) #' sc <- Normalizedata( diff --git a/R/DIscBIO-generic-plotexptsneMB.R b/R/DIscBIO-generic-plotexptsneMB.R index e3aa0d0..fc3ca35 100644 --- a/R/DIscBIO-generic-plotexptsneMB.R +++ b/R/DIscBIO-generic-plotexptsneMB.R @@ -11,7 +11,7 @@ #' NULL and the first element of \code{g} is chosen. #' @return t-SNE plot for one particular gene #' @examples -#' \dontrun{ +#' \donttest{ #' sc <- DISCBIO(valuesG1msReduced) #' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) #' sc <- Normalizedata( diff --git a/R/MB_Order.R b/R/MB_Order.R index 00805e1..6e7c54b 100644 --- a/R/MB_Order.R +++ b/R/MB_Order.R @@ -9,7 +9,7 @@ #' @importFrom TSCAN TSCANorder #' @return The DISCBIO-class object input with the MBordering slot filled. #' @examples -#' \dontrun{ +#' \donttest{ #' sc <- DISCBIO(valuesG1msReduced) #' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) #' sc <- Normalizedata( diff --git a/R/NetAnalysis.R b/R/NetAnalysis.R index 0083f4d..1bc5d31 100644 --- a/R/NetAnalysis.R +++ b/R/NetAnalysis.R @@ -12,7 +12,7 @@ #' @importFrom NetIndices GenInd #' @return A network analysis table #' @examples -#' \dontrun{ +#' \donttest{ #' sc <- DISCBIO(valuesG1msReduced) #' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) #' sc <- Normalizedata( diff --git a/R/Networking.R b/R/Networking.R index 609617e..c564c46 100644 --- a/R/Networking.R +++ b/R/Networking.R @@ -13,7 +13,7 @@ #' @importFrom graphics plot rasterImage #' @return A plot of the network #' @examples -#' \dontrun{ +#' \donttest{ #' sc <- DISCBIO(valuesG1msReduced) #' sc <- NoiseFiltering(sc, export=FALSE) #' sc <- Normalizedata( diff --git a/R/PPI.R b/R/PPI.R index 724eabc..ae2d77e 100644 --- a/R/PPI.R +++ b/R/PPI.R @@ -11,7 +11,7 @@ #' @return A TSV file stored in the user's file system and its corresponding #' `data.frame` object in R. #' @examples -#' \dontrun{ +#' \donttest{ #' sc <- DISCBIO(valuesG1msReduced) #' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) #' sc <- Normalizedata( diff --git a/R/VolcanoPlot.R b/R/VolcanoPlot.R index 53201f0..d48de62 100644 --- a/R/VolcanoPlot.R +++ b/R/VolcanoPlot.R @@ -18,7 +18,7 @@ #' @return A volcano plot #' @export #' @examples -#' \dontrun{ +#' \donttest{ #' sc <- DISCBIO(valuesG1msReduced) #' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) #' sc <- Normalizedata( diff --git a/man/DEGanalysis.Rd b/man/DEGanalysis.Rd index 4f641eb..9339b6d 100644 --- a/man/DEGanalysis.Rd +++ b/man/DEGanalysis.Rd @@ -70,6 +70,7 @@ sc <- Clustexp(sc, cln=3) # K-means clustering sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) sc <- comptSNE(sc, max_iter=100) DEGanalysis( - sc, Clustering="K-means", K=3, fdr=0.1, name="Name", export = FALSE + sc, Clustering="K-means", K=3, fdr=0.1, name="Name", export = FALSE, + nresamp=5, nperms=20 ) } diff --git a/man/J48DT.Rd b/man/J48DT.Rd index dcef16e..5fabfcc 100644 --- a/man/J48DT.Rd +++ b/man/J48DT.Rd @@ -34,7 +34,8 @@ sc <- Clustexp(sc, cln=3) # K-means clustering sc <- comptSNE(sc, max_iter=100) cdiff <- DEGanalysis2clust( sc, Clustering="K-means", K=3, fdr=.2, name="Name", First="CL1", - Second="CL2", export=FALSE + Second="CL2", export=FALSE, + nresamp=5, nperms=20 ) sigDEG <- cdiff[[1]] DATAforDT <- ClassVectoringDT( diff --git a/man/MBClustDiffGenes.Rd b/man/MBClustDiffGenes.Rd index a1e8e08..4175937 100644 --- a/man/MBClustDiffGenes.Rd +++ b/man/MBClustDiffGenes.Rd @@ -44,7 +44,7 @@ A list containing two tables. ClustDiffGenes } \examples{ -\dontrun{ +\donttest{ sc <- DISCBIO(valuesG1msReduced) sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) sc <- Normalizedata( diff --git a/man/MB_Order.Rd b/man/MB_Order.Rd index 525c985..5271ddb 100644 --- a/man/MB_Order.Rd +++ b/man/MB_Order.Rd @@ -22,7 +22,7 @@ This function takes the exact output of exprmclust function and connects cluster centers. } \examples{ -\dontrun{ +\donttest{ sc <- DISCBIO(valuesG1msReduced) sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) sc <- Normalizedata( diff --git a/man/MBclustheatmap.Rd b/man/MBclustheatmap.Rd index 16ccd24..6c7bcf9 100644 --- a/man/MBclustheatmap.Rd +++ b/man/MBclustheatmap.Rd @@ -31,7 +31,7 @@ This functions plots a heatmap of the distance matrix grouped the x and y-axes. } \examples{ -\dontrun{ +\donttest{ sc <- DISCBIO(valuesG1msReduced) sc <- NoiseFiltering(sc, export=FALSE) sc <- Normalizedata( diff --git a/man/NetAnalysis.Rd b/man/NetAnalysis.Rd index 3134f07..55812b5 100644 --- a/man/NetAnalysis.Rd +++ b/man/NetAnalysis.Rd @@ -23,7 +23,7 @@ This function checks the connectivity degree and the betweenness networks } \examples{ -\dontrun{ +\donttest{ sc <- DISCBIO(valuesG1msReduced) sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) sc <- Normalizedata( diff --git a/man/Networking.Rd b/man/Networking.Rd index 2ad8da2..93d5bf6 100644 --- a/man/Networking.Rd +++ b/man/Networking.Rd @@ -25,7 +25,7 @@ A plot of the network This function uses STRING-api to plot the network. } \examples{ -\dontrun{ +\donttest{ sc <- DISCBIO(valuesG1msReduced) sc <- NoiseFiltering(sc, export=FALSE) sc <- Normalizedata( diff --git a/man/PPI.Rd b/man/PPI.Rd index f73ef94..eeb1078 100644 --- a/man/PPI.Rd +++ b/man/PPI.Rd @@ -23,7 +23,7 @@ This function uses STRING-api. The outcome of STRING analysis will be stored in tab separated values (TSV) files. } \examples{ -\dontrun{ +\donttest{ sc <- DISCBIO(valuesG1msReduced) sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) sc <- Normalizedata( diff --git a/man/VolcanoPlot.Rd b/man/VolcanoPlot.Rd index 6377c89..9ece925 100644 --- a/man/VolcanoPlot.Rd +++ b/man/VolcanoPlot.Rd @@ -29,7 +29,7 @@ Plotting differentially expressed genes (DEGs) in a particular significance versus fold-change on the y and x axes, respectively. } \examples{ -\dontrun{ +\donttest{ sc <- DISCBIO(valuesG1msReduced) sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) sc <- Normalizedata( diff --git a/man/plotexptsneMB.Rd b/man/plotexptsneMB.Rd index 7d03cec..e71ed2c 100644 --- a/man/plotexptsneMB.Rd +++ b/man/plotexptsneMB.Rd @@ -29,7 +29,7 @@ The t-SNE map representation can also be used to analyze gene expression patterns } \examples{ -\dontrun{ +\donttest{ sc <- DISCBIO(valuesG1msReduced) sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) sc <- Normalizedata( From 717d408099b9f15c88802a00e33aec283759ab06 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 29 Jun 2020 11:19:14 +0200 Subject: [PATCH 13/99] Added "don't export" option to PPI() --- R/PPI.R | 8 +++++--- man/PPI.Rd | 4 ++-- 2 files changed, 7 insertions(+), 5 deletions(-) diff --git a/R/PPI.R b/R/PPI.R index ae2d77e..bc2da95 100644 --- a/R/PPI.R +++ b/R/PPI.R @@ -4,7 +4,7 @@ #' @export #' @param data A gene list. #' @param FileName A string vector showing the name to be used to save the -#' resulted table. +#' resulted table. If null, no file will be exported #' @param species The taxonomy name/id. Default is "9606" for Homo sapiens. #' @importFrom httr content #' @importFrom readr read_tsv @@ -28,7 +28,7 @@ #' FileName <- paste0(DEGs) #' PPI(data, FileName) #' } -PPI <- function(data, FileName, species = "9606") { +PPI <- function(data, FileName = NULL, species = "9606") { # Save base enpoint as variable string_api_url <- "https://string-db.org/api/" output_format <- "tsv" #"json", "tsv-no-header", "tsv", "xml" @@ -60,6 +60,8 @@ PPI <- function(data, FileName, species = "9606") { # Process API request content repo_content <- content(repos) results <- read_tsv(repo_content) - write.csv(results, file = paste0("PPI-", FileName, ".csv")) + if (!is.null(FileName)) { + write.csv(results, file = paste0("PPI-", FileName, ".csv")) + } return(results) } \ No newline at end of file diff --git a/man/PPI.Rd b/man/PPI.Rd index eeb1078..e3da68e 100644 --- a/man/PPI.Rd +++ b/man/PPI.Rd @@ -4,13 +4,13 @@ \alias{PPI} \title{Defining protein-protein interactions (PPI) over a list of genes,} \usage{ -PPI(data, FileName, species = "9606") +PPI(data, FileName = NULL, species = "9606") } \arguments{ \item{data}{A gene list.} \item{FileName}{A string vector showing the name to be used to save the -resulted table.} +resulted table. If null, no file will be exported} \item{species}{The taxonomy name/id. Default is "9606" for Homo sapiens.} } From 499a021863a11afde16580ee4fc2c9f8088aa916 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 29 Jun 2020 11:35:22 +0200 Subject: [PATCH 14/99] Added year to cited publication on DESCRIPTION --- DESCRIPTION | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/DESCRIPTION b/DESCRIPTION index 68d1310..a05b554 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -33,7 +33,7 @@ Description: An open, multi-algorithmic pipeline for easy, fast and efficient characterize them. The pipeline consists of four successive steps: data pre-processing, cellular clustering with pseudo-temporal ordering, defining differential expressed genes and biomarker identification. This package - implements extensions of the work published by Ghannoum et. al. + implements extensions of the work published by Ghannoum et. al. (2019) . License: MIT + file LICENSE Encoding: UTF-8 From 39f95243c56134736693b4d87bf20fc514ccf48f Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 29 Jun 2020 11:58:17 +0200 Subject: [PATCH 15/99] Minor doc updates --- DESCRIPTION | 2 +- tests/testthat/test.integration.R | 2 +- 2 files changed, 2 insertions(+), 2 deletions(-) diff --git a/DESCRIPTION b/DESCRIPTION index a05b554..ca2cff2 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -52,7 +52,7 @@ Suggests: BiocManager VignetteBuilder: knitr LazyData: true -RoxygenNote: 7.1.0 +RoxygenNote: 7.1.1 BugReports: https://github.com/ocbe-uio/DIscBIO/issues Collate: 'DIscBIO-classes.R' diff --git a/tests/testthat/test.integration.R b/tests/testthat/test.integration.R index df48a5c..0aaba8c 100644 --- a/tests/testthat/test.integration.R +++ b/tests/testthat/test.integration.R @@ -5,7 +5,7 @@ context("Converting other formats to DISCBIO") # ------------------------------------------------------------------------------ # Setting up datasets # ------------------------------------------------------------------------------ -pmbc_seurat <- Seurat::pbmc_small +pmbc_seurat <- Seurat::pbmc_small # TODO: replace with library(Seurat); data(...) to better pass r-devel? pmbc_sce <- Seurat::as.SingleCellExperiment(pmbc_seurat) g1_sce <- SingleCellExperiment::SingleCellExperiment( list(counts=as.matrix(valuesG1msReduced)) From 9f2ba9a8ccc26831ac883a487c9fe0090845c668 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 29 Jun 2020 11:58:49 +0200 Subject: [PATCH 16/99] Expanded output of DEGanalysis2clust --- R/DIscBIO-generic-DEGanalysis2clust.R | 13 +++++++++++-- 1 file changed, 11 insertions(+), 2 deletions(-) diff --git a/R/DIscBIO-generic-DEGanalysis2clust.R b/R/DIscBIO-generic-DEGanalysis2clust.R index 18c01af..fb7ea66 100644 --- a/R/DIscBIO-generic-DEGanalysis2clust.R +++ b/R/DIscBIO-generic-DEGanalysis2clust.R @@ -171,6 +171,7 @@ setMethod( "Up-regulated-", name, First, "in", First, "VS", Second, ".csv" ) + FinalDEGsL <- NULL if (length(FDRl) > 0) { genes <- siggenes.table$genes.lo[, 3] if (quiet) { @@ -239,7 +240,7 @@ setMethod( DEGsS <- c(DEGsS, FinalDEGsL[, 2]) DEGsE <- c(DEGsE, as.character(FinalDEGsL[, 3])) } - + FinalDEGsU <- NULL if (length(FDRu) > 0) { genes <- siggenes.table$genes.up[, 3] if (quiet) { @@ -335,6 +336,14 @@ setMethod( write.csv(DEGsTable, file = "DEGsTable.csv") write.csv(sigDEG, file = "sigDEG.csv") } - return(list(sigDEG, DEGsTable)) + return( + list( + sigDEG = sigDEG, + DEGsTable = DEGsTable, + FinalDEGsL = FinalDEGsL, + FinalDEGsU = FinalDEGsU + + ) + ) } ) \ No newline at end of file From 9216eb840d2af8eddc17fc8ee003227034ef69bc Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 29 Jun 2020 11:59:09 +0200 Subject: [PATCH 17/99] Improved examples --- R/NetAnalysis.R | 16 ++++++---------- R/PPI.R | 21 +++++++++------------ man/NetAnalysis.Rd | 16 ++++++---------- man/PPI.Rd | 19 ++++++++----------- 4 files changed, 29 insertions(+), 43 deletions(-) diff --git a/R/NetAnalysis.R b/R/NetAnalysis.R index 1bc5d31..3b9be6a 100644 --- a/R/NetAnalysis.R +++ b/R/NetAnalysis.R @@ -19,16 +19,12 @@ #' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, #' dsn=1, rseed=17000 #' ) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF") -#' sc <- Clustexp(sc, cln=3) # K-means clustering -#' sc <- comptSNE(sc, max_iter=100) -#' dff <- DEGanalysis2clust(sc, Clustering="K-means", K=3, fdr=0.1, name="Name") -#' DEGs <- dff[[2]][1, 6] -#' data <- read.csv(file=paste0(DEGs),head=TRUE,sep=",") -#' data <- data[,3] -#' FileName <- paste0(DEGs) -#' ppi <- PPI(data, FileName) -#' NetAnalysis(ppi) +#' sc <- FinalPreprocessing(sc, export=FALSE, quiet=TRUE) +#' sc <- Clustexp(sc, cln=3, quiet=TRUE) # K-means clustering +#' sc <- comptSNE(sc, max_iter=100, quiet=TRUE) +#' dff <- DEGanalysis2clust(sc, K=3, export=FALSE, quiet=TRUE, plot=FALSE) +#' ppi <- PPI(dff$FinalDEGsU[, "Gene Name"]) +#' NetAnalysis(ppi, export=FALSE) #' } NetAnalysis <- function(data, export = TRUE, FileName = "1") { if (length(data[, 1]) < 1) diff --git a/R/PPI.R b/R/PPI.R index bc2da95..f61b61f 100644 --- a/R/PPI.R +++ b/R/PPI.R @@ -8,8 +8,9 @@ #' @param species The taxonomy name/id. Default is "9606" for Homo sapiens. #' @importFrom httr content #' @importFrom readr read_tsv -#' @return A TSV file stored in the user's file system and its corresponding -#' `data.frame` object in R. +#' @return Either a TSV file stored in the user's file system and its +#' corresponding `data.frame` object in R or and R object containing that +#' information. #' @examples #' \donttest{ #' sc <- DISCBIO(valuesG1msReduced) @@ -18,15 +19,11 @@ #' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, #' dsn=1, rseed=17000 #' ) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF") -#' sc <- Clustexp(sc, cln=3) # K-means clustering -#' sc <- comptSNE(sc, max_iter=100) -#' dff <- DEGanalysis2clust(sc, Clustering="K-means", K=3, fdr=0.1, name="Name") -#' DEGs <- dff[[2]][1, 6] -#' data <- read.csv(file=paste0(DEGs),head=TRUE,sep=",") -#' data <- data[,3] -#' FileName <- paste0(DEGs) -#' PPI(data, FileName) +#' sc <- FinalPreprocessing(sc, export=FALSE, quiet=TRUE) +#' sc <- Clustexp(sc, cln=3, quiet=TRUE) # K-means clustering +#' sc <- comptSNE(sc, max_iter=100, quiet=TRUE) +#' dff <- DEGanalysis2clust(sc, K=3, export=FALSE, quiet=TRUE, plot=FALSE) +#' PPI(dff$FinalDEGsU[, "Gene Name"]) #' } PPI <- function(data, FileName = NULL, species = "9606") { # Save base enpoint as variable @@ -54,7 +51,7 @@ PPI <- function(data, FileName = NULL, species = "9606") { "Examine response components =", status_code(repos), "\t", - "200 means successful", + "(200 means successful)", "\n" ) # Process API request content diff --git a/man/NetAnalysis.Rd b/man/NetAnalysis.Rd index 55812b5..109018b 100644 --- a/man/NetAnalysis.Rd +++ b/man/NetAnalysis.Rd @@ -30,15 +30,11 @@ sc <- Normalizedata( sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, dsn=1, rseed=17000 ) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF") -sc <- Clustexp(sc, cln=3) # K-means clustering -sc <- comptSNE(sc, max_iter=100) -dff <- DEGanalysis2clust(sc, Clustering="K-means", K=3, fdr=0.1, name="Name") -DEGs <- dff[[2]][1, 6] -data <- read.csv(file=paste0(DEGs),head=TRUE,sep=",") -data <- data[,3] -FileName <- paste0(DEGs) -ppi <- PPI(data, FileName) -NetAnalysis(ppi) +sc <- FinalPreprocessing(sc, export=FALSE, quiet=TRUE) +sc <- Clustexp(sc, cln=3, quiet=TRUE) # K-means clustering +sc <- comptSNE(sc, max_iter=100, quiet=TRUE) +dff <- DEGanalysis2clust(sc, K=3, export=FALSE, quiet=TRUE, plot=FALSE) +ppi <- PPI(dff$FinalDEGsU[, "Gene Name"]) +NetAnalysis(ppi, export=FALSE) } } diff --git a/man/PPI.Rd b/man/PPI.Rd index e3da68e..9646c95 100644 --- a/man/PPI.Rd +++ b/man/PPI.Rd @@ -15,8 +15,9 @@ resulted table. If null, no file will be exported} \item{species}{The taxonomy name/id. Default is "9606" for Homo sapiens.} } \value{ -A TSV file stored in the user's file system and its corresponding - `data.frame` object in R. +Either a TSV file stored in the user's file system and its +corresponding `data.frame` object in R or and R object containing that +information. } \description{ This function uses STRING-api. The outcome of STRING analysis @@ -30,14 +31,10 @@ sc <- Normalizedata( sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, dsn=1, rseed=17000 ) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF") -sc <- Clustexp(sc, cln=3) # K-means clustering -sc <- comptSNE(sc, max_iter=100) -dff <- DEGanalysis2clust(sc, Clustering="K-means", K=3, fdr=0.1, name="Name") -DEGs <- dff[[2]][1, 6] -data <- read.csv(file=paste0(DEGs),head=TRUE,sep=",") -data <- data[,3] -FileName <- paste0(DEGs) -PPI(data, FileName) +sc <- FinalPreprocessing(sc, export=FALSE, quiet=TRUE) +sc <- Clustexp(sc, cln=3, quiet=TRUE) # K-means clustering +sc <- comptSNE(sc, max_iter=100, quiet=TRUE) +dff <- DEGanalysis2clust(sc, K=3, export=FALSE, quiet=TRUE, plot=FALSE) +PPI(dff$FinalDEGsU[, "Gene Name"]) } } From a8527b2b40f096eafc1865f382ff10cd21190da0 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 29 Jun 2020 13:28:49 +0200 Subject: [PATCH 18/99] Added a "don't export* switch to Networking --- R/Networking.R | 40 +++++++++++++++++++++------------------- man/Networking.Rd | 30 +++++++++++++++--------------- 2 files changed, 36 insertions(+), 34 deletions(-) diff --git a/R/Networking.R b/R/Networking.R index c564c46..cbbfeac 100644 --- a/R/Networking.R +++ b/R/Networking.R @@ -3,7 +3,8 @@ #' @export #' @param data A gene list. #' @param FileName A string vector showing the name to be used to save the -#' resulted network. +#' resulted network. If `NULL`, the network will be saved to a temporary +#' directory #' @param species The taxonomy name/id. Default is "9606" for Homo sapiens. #' @param plot_width Plot width #' @param plot_height Plot height @@ -15,26 +16,23 @@ #' @examples #' \donttest{ #' sc <- DISCBIO(valuesG1msReduced) -#' sc <- NoiseFiltering(sc, export=FALSE) +#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE, plot=FALSE) #' sc <- Normalizedata( #' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, #' dsn=1, rseed=17000 #' ) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF") -#' sc <- Clustexp(sc, cln=3) # K-means clustering -#' sc <- comptSNE(sc, max_iter=100) -#' dff <- DEGanalysis2clust(sc, Clustering="K-means", K=3, fdr=0.1, name="Name") -#' DEGs <- dff[[2]][1, 6] -#' data <- read.csv(file=paste0(DEGs),head=TRUE,sep=",") -#' data <- data[,3] -#' FileName <- paste0(DEGs) -#' ppi <- PPI(data, FileName) -#' networking <- NetAnalysis(ppi) -#' FileName <- "Up.DownDEG" -#' Networking(data, FileName) +#' sc <- FinalPreprocessing(sc, export=FALSE, quiet=TRUE) +#' sc <- Clustexp(sc, cln=3, quiet=TRUE) #' K-means clustering +#' sc <- comptSNE(sc, max_iter=100, quiet=TRUE) +#' dff <- DEGanalysis2clust(sc, K=3, export=FALSE, quiet=TRUE, plot=FALSE) +#' Networking(dff$FinalDEGsU[, "Gene Name"]) #' } Networking <- function( - data, FileName, species = "9606", plot_width = 25, plot_height = 15 + data, + FileName = NULL, + species = "9606", + plot_width = 25, + plot_height = 15 ) { if (length(data) > 600) { @@ -66,10 +64,14 @@ Networking <- function( "Examine response components =", status_code(repos), "\t", "200 means successful", "\n" ) - y = repos$request$url - download.file(y, paste0("network", FileName, ".png"), mode = 'wb') - Network <- - readPNG(paste0("network", FileName, ".png"), native = TRUE) + y <- repos$request$url + if (!is.null(FileName)) { + FileName <- paste0("network", FileName, ".png") + } else { + FileName <- tempfile() + } + download.file(y, FileName, mode = 'wb') + Network <- readPNG(FileName, native=TRUE) set_plot_dimensions <- function(width_choice, height_choice) { options(repr.plot.width = width_choice, diff --git a/man/Networking.Rd b/man/Networking.Rd index 93d5bf6..6915731 100644 --- a/man/Networking.Rd +++ b/man/Networking.Rd @@ -4,13 +4,20 @@ \alias{Networking} \title{Plotting the network.} \usage{ -Networking(data, FileName, species = "9606", plot_width = 25, plot_height = 15) +Networking( + data, + FileName = NULL, + species = "9606", + plot_width = 25, + plot_height = 15 +) } \arguments{ \item{data}{A gene list.} \item{FileName}{A string vector showing the name to be used to save the -resulted network.} +resulted network. If `NULL`, the network will be saved to a temporary +directory} \item{species}{The taxonomy name/id. Default is "9606" for Homo sapiens.} @@ -27,22 +34,15 @@ This function uses STRING-api to plot the network. \examples{ \donttest{ sc <- DISCBIO(valuesG1msReduced) -sc <- NoiseFiltering(sc, export=FALSE) +sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE, plot=FALSE) sc <- Normalizedata( sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, dsn=1, rseed=17000 ) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF") -sc <- Clustexp(sc, cln=3) # K-means clustering -sc <- comptSNE(sc, max_iter=100) -dff <- DEGanalysis2clust(sc, Clustering="K-means", K=3, fdr=0.1, name="Name") -DEGs <- dff[[2]][1, 6] -data <- read.csv(file=paste0(DEGs),head=TRUE,sep=",") -data <- data[,3] -FileName <- paste0(DEGs) -ppi <- PPI(data, FileName) -networking <- NetAnalysis(ppi) -FileName <- "Up.DownDEG" -Networking(data, FileName) +sc <- FinalPreprocessing(sc, export=FALSE, quiet=TRUE) +sc <- Clustexp(sc, cln=3, quiet=TRUE) #' K-means clustering +sc <- comptSNE(sc, max_iter=100, quiet=TRUE) +dff <- DEGanalysis2clust(sc, K=3, export=FALSE, quiet=TRUE, plot=FALSE) +Networking(dff$FinalDEGsU[, "Gene Name"]) } } From 8f8bb70d008652d8785c7efbf99d79044b7615a0 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 29 Jun 2020 14:42:24 +0200 Subject: [PATCH 19/99] Added functions to reformat SAMR's siggenes tables --- R/reformatSiggenes.R | 28 ++++++++++++++++++++++++++++ R/replaceDecimals.R | 8 ++++++++ man/reformatSiggenes.Rd | 14 ++++++++++++++ man/replaceDecimals.Rd | 17 +++++++++++++++++ 4 files changed, 67 insertions(+) create mode 100644 R/reformatSiggenes.R create mode 100644 R/replaceDecimals.R create mode 100644 man/reformatSiggenes.Rd create mode 100644 man/replaceDecimals.Rd diff --git a/R/reformatSiggenes.R b/R/reformatSiggenes.R new file mode 100644 index 0000000..6d60a28 --- /dev/null +++ b/R/reformatSiggenes.R @@ -0,0 +1,28 @@ +#' @title Reformat Siggenes Table +#' @description Reformats the Siggenes table output from the SAMR package +#' @seealso replaceDecimals +reformatSiggenes <- function(table) { + if (is.null(table)) return(table) + table <- as.data.frame(table) + # ========================================================================== + # Replacing decimal separators + # ========================================================================== + table[, "Score(d)"] <- replaceDecimals(table[, "Score(d)"]) + table[, "Numerator(r)"] <- replaceDecimals(table[, "Numerator(r)"]) + table[, "Denominator(s+s0)"] <- replaceDecimals(table[, "Denominator(s+s0)"]) + table[, "Fold Change"] <- replaceDecimals(table[, "Fold Change"]) + table[, "q-value(%)"] <- replaceDecimals(table[, "q-value(%)"]) + # ========================================================================== + # Changing vector classes + # ========================================================================== + table[, "Row"] <- as.numeric(table[, "Row"]) + table[, "Score(d)"] <- as.numeric(table[, "Score(d)"]) + table[, "Numerator(r)"] <- as.numeric(table[, "Numerator(r)"]) + table[, "Denominator(s+s0)"] <- as.numeric(table[, "Denominator(s+s0)"]) + table[, "Fold Change"] <- as.numeric(table[, "Fold Change"]) + table[, "q-value(%)"] <- as.numeric(table[, "q-value(%)"]) + # ========================================================================== + # Returning output + # ========================================================================== + return(table) +} \ No newline at end of file diff --git a/R/replaceDecimals.R b/R/replaceDecimals.R new file mode 100644 index 0000000..dd50bd2 --- /dev/null +++ b/R/replaceDecimals.R @@ -0,0 +1,8 @@ +#' @title Replace Decimals +#' @description Replaces decimals separators between comma and periods +#' @note This function was especially designed to be used with retormatSiggenes +#' @seealso reformatSiggenes +replaceDecimals <- function(x, from=",", to=".") { + x <- gsub(",", ".", x) + return(x) +} \ No newline at end of file diff --git a/man/reformatSiggenes.Rd b/man/reformatSiggenes.Rd new file mode 100644 index 0000000..8202e81 --- /dev/null +++ b/man/reformatSiggenes.Rd @@ -0,0 +1,14 @@ +% Generated by roxygen2: do not edit by hand +% Please edit documentation in R/reformatSiggenes.R +\name{reformatSiggenes} +\alias{reformatSiggenes} +\title{Reformat Siggenes Table} +\usage{ +reformatSiggenes(table) +} +\description{ +Reformats the Siggenes table output from the SAMR package +} +\seealso{ +replaceDecimals +} diff --git a/man/replaceDecimals.Rd b/man/replaceDecimals.Rd new file mode 100644 index 0000000..074ce27 --- /dev/null +++ b/man/replaceDecimals.Rd @@ -0,0 +1,17 @@ +% Generated by roxygen2: do not edit by hand +% Please edit documentation in R/replaceDecimals.R +\name{replaceDecimals} +\alias{replaceDecimals} +\title{Replace Decimals} +\usage{ +replaceDecimals(x, from = ",", to = ".") +} +\description{ +Replaces decimals separators between comma and periods +} +\note{ +This function was especially designed to be used with retormatSiggenes +} +\seealso{ +reformatSiggenes +} From 1ee96f3fee14479c89f4c0ecf2bf65373df7203a Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 29 Jun 2020 15:55:42 +0200 Subject: [PATCH 20/99] Improved output --- R/DIscBIO-generic-DEGanalysis2clust.R | 9 +++++++-- 1 file changed, 7 insertions(+), 2 deletions(-) diff --git a/R/DIscBIO-generic-DEGanalysis2clust.R b/R/DIscBIO-generic-DEGanalysis2clust.R index fb7ea66..bb1bb77 100644 --- a/R/DIscBIO-generic-DEGanalysis2clust.R +++ b/R/DIscBIO-generic-DEGanalysis2clust.R @@ -140,6 +140,11 @@ setMethod( siggenes.table <- samr.compute.siggenes.table( samr.obj, delta, data, delta.table ) + # ------------------------------------------------------------------ + # Reformat siggenes.table as data.frame + # ------------------------------------------------------------------ + siggenes.table$genes.lo <- reformatSiggenes(siggenes.table$genes.lo) + siggenes.table$genes.up <- reformatSiggenes(siggenes.table$genes.up) FDRl <- as.numeric(siggenes.table$genes.lo[, 8]) / 100 FDRu <- as.numeric(siggenes.table$genes.up[, 8]) / 100 @@ -171,7 +176,7 @@ setMethod( "Up-regulated-", name, First, "in", First, "VS", Second, ".csv" ) - FinalDEGsL <- NULL + FinalDEGsL <- data.frame() if (length(FDRl) > 0) { genes <- siggenes.table$genes.lo[, 3] if (quiet) { @@ -240,7 +245,7 @@ setMethod( DEGsS <- c(DEGsS, FinalDEGsL[, 2]) DEGsE <- c(DEGsE, as.character(FinalDEGsL[, 3])) } - FinalDEGsU <- NULL + FinalDEGsU <- data.frame() if (length(FDRu) > 0) { genes <- siggenes.table$genes.up[, 3] if (quiet) { From bc50a9b44be8db598e2ea905240a4cfdee35e36c Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 29 Jun 2020 15:56:34 +0200 Subject: [PATCH 21/99] Fixed VolcanoPlot behavior and example --- R/VolcanoPlot.R | 31 ++++++++++++++++--------------- man/VolcanoPlot.Rd | 17 +++++++---------- 2 files changed, 23 insertions(+), 25 deletions(-) diff --git a/R/VolcanoPlot.R b/R/VolcanoPlot.R index d48de62..4b73f58 100644 --- a/R/VolcanoPlot.R +++ b/R/VolcanoPlot.R @@ -8,8 +8,7 @@ #' Default is 0.05 #' @param fc A numeric value of the fold change. Default is 0.5. #' @param FS A numeric value of the font size. Default is 0.4. -#' @param name A string vector showing the name to be used to save the resulted -#' tables. +#' @param name A string vector showing the name to be used on the plot title #' @importFrom samr samr samr.compute.delta.table samr.plot #' samr.compute.siggenes.table #' @importFrom graphics title @@ -20,28 +19,30 @@ #' @examples #' \donttest{ #' sc <- DISCBIO(valuesG1msReduced) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) +#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE, plot=FALSE) #' sc <- Normalizedata( #' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, #' dsn=1, rseed=17000 #' ) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF") -#' sc <- Clustexp(sc, cln=3) # K-means clustering -#' sc <- comptSNE(sc, max_iter=100) -#' dff <- DEGanalysis2clust(sc, Clustering="K-means", K=3, fdr=0.1, name="Name") -#' name <- dff[[2]][1, 6] -#' U <- read.csv(file = paste0(name), head=TRUE, sep=",") -#' VolcanoPlot(U, value=0.05, name=name, adj=FALSE, FS=.4) +#' sc <- FinalPreprocessing(sc, export=FALSE, quiet=TRUE) +#' sc <- Clustexp(sc, cln=3, quiet=TRUE) # K-means clustering +#' sc <- comptSNE(sc, max_iter=100, quiet=TRUE) +#' dff <- DEGanalysis2clust(sc, K=3, export=FALSE, quiet=TRUE, plot=FALSE) +#' VolcanoPlot(dff$FinalDEGsU, value=0.05, FS=.4) #' } -VolcanoPlot <- function(object, value = 0.05, name, fc = 0.5, FS = .4) { +VolcanoPlot <- function(object, value = 0.05, name = NULL, fc = 0.5, FS = .4) { if (length(object[1, ]) > 8) { object <- object[, -1] } NO0 <- object[, 8] - NO0 <- NO0[-which(NO0 == 0)] + NO0 <- NO0[which(NO0 != 0)] w <- which.min(NO0) adjV <- NO0[w] / 100 - object[, 8] <- ifelse(object[, 8] == 0, adjV, object[, 8]) + object[, 8] <- ifelse(object[, 8] == 0 & length(adjV) > 0, adjV, object[, 8]) + if (all(object[, 8] == 0)) { + message("All q-values are 0. Adjusting") + object[, 8] <- object[, 8] + 1e-10 + } with( object, plot( @@ -52,12 +53,12 @@ VolcanoPlot <- function(object, value = 0.05, name, fc = 0.5, FS = .4) { las = 1, xlab = "log2 Fold Change", ylab = "-log10 FDR", - sub = paste0("Volcano plot ", name), + sub = paste("Volcano plot", name), font.sub = 4, col.sub = "black" ) ) - FC <- subset(object, abs(object[, 7]) > fc) # Fold Change + FC <- subset(object, abs(object[, 7]) > fc) # Fold Change sigFC <- subset( object, object[, 8] < value & abs(object[, 7]) > fc ) # Significant genes diff --git a/man/VolcanoPlot.Rd b/man/VolcanoPlot.Rd index 9ece925..09b48f1 100644 --- a/man/VolcanoPlot.Rd +++ b/man/VolcanoPlot.Rd @@ -13,8 +13,7 @@ in a particular cluster} \item{value}{A numeric value of the false discovery rate. Default is 0.05.. Default is 0.05} -\item{name}{A string vector showing the name to be used to save the resulted -tables.} +\item{name}{A string vector showing the name to be used on the plot title} \item{fc}{A numeric value of the fold change. Default is 0.5.} @@ -31,17 +30,15 @@ Plotting differentially expressed genes (DEGs) in a particular \examples{ \donttest{ sc <- DISCBIO(valuesG1msReduced) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) +sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE, plot=FALSE) sc <- Normalizedata( sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, dsn=1, rseed=17000 ) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF") -sc <- Clustexp(sc, cln=3) # K-means clustering -sc <- comptSNE(sc, max_iter=100) -dff <- DEGanalysis2clust(sc, Clustering="K-means", K=3, fdr=0.1, name="Name") -name <- dff[[2]][1, 6] -U <- read.csv(file = paste0(name), head=TRUE, sep=",") -VolcanoPlot(U, value=0.05, name=name, adj=FALSE, FS=.4) +sc <- FinalPreprocessing(sc, export=FALSE, quiet=TRUE) +sc <- Clustexp(sc, cln=3, quiet=TRUE) # K-means clustering +sc <- comptSNE(sc, max_iter=100, quiet=TRUE) +dff <- DEGanalysis2clust(sc, K=3, export=FALSE, quiet=TRUE, plot=FALSE) +VolcanoPlot(dff$FinalDEGsU, value=0.05, FS=.4) } } From e6d4f576bc8f724cc643fd8f1220b50e2bcf4d34 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 29 Jun 2020 16:09:45 +0200 Subject: [PATCH 22/99] Improved documentation --- DESCRIPTION | 98 +++++++++++++++++++++-------------------- R/reformatSiggenes.R | 1 + R/replaceDecimals.R | 6 ++- man/VolcanoPlot.Rd | 2 +- man/reformatSiggenes.Rd | 3 ++ man/replaceDecimals.Rd | 10 ++++- 6 files changed, 69 insertions(+), 51 deletions(-) diff --git a/DESCRIPTION b/DESCRIPTION index ca2cff2..e875085 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -55,51 +55,53 @@ LazyData: true RoxygenNote: 7.1.1 BugReports: https://github.com/ocbe-uio/DIscBIO/issues Collate: - 'DIscBIO-classes.R' - 'DIscBIO-generic-ClassVectoringDT.R' - 'DIscBIO-generic-Clustexp.R' - 'DIscBIO-generic-DEGanalysis.R' - 'DIscBIO-generic-DEGanalysis2clust.R' - 'DIscBIO-generic-Exprmclust.R' - 'DIscBIO-generic-FinalPreprocessing.R' - 'DIscBIO-generic-FindOutliersKM.R' - 'DIscBIO-generic-FindOutliersMB.R' - 'DIscBIO-generic-KMClustDiffGenes.R' - 'DIscBIO-generic-KMclustheatmap.R' - 'DIscBIO-generic-KmeanOrder.R' - 'DIscBIO-generic-MBClustDiffGenes.R' - 'DIscBIO-generic-MBclustheatmap.R' - 'DIscBIO-generic-NoiseFiltering.R' - 'DIscBIO-generic-Normalizedata.R' - 'DIscBIO-generic-PCAplotSymbols.R' - 'DIscBIO-generic-PlotmclustMB.R' - 'DIscBIO-generic-comptSNE.R' - 'DIscBIO-generic-comptsneMB.R' - 'DIscBIO-generic-plotExptSNE.R' - 'DIscBIO-generic-plotGap.R' - 'DIscBIO-generic-plotKmeansLabelstSNE.R' - 'DIscBIO-generic-plotMBLabelstSNE.R' - 'DIscBIO-generic-plotOrderKMtsne.R' - 'DIscBIO-generic-plotOrderMBtsne.R' - 'DIscBIO-generic-plotSilhouette.R' - 'DIscBIO-generic-plotSymbolstSNE.R' - 'DIscBIO-generic-plotexptsneMB.R' - 'DIscBIO-generic-plotsilhouetteMB.R' - 'DIscBIO-generic-plottSNE.R' - 'DIscBIO-generic-plottsneMB.R' - 'J48DT.R' - 'J48DTeval.R' - 'Jaccard.R' - 'MB_Order.R' - 'NetAnalysis.R' - 'Networking.R' - 'PPI.R' - 'PlotMBexpPCA.R' - 'PlotMBorderPCA.R' - 'RpartDT.R' - 'RpartEVAL.R' - 'VolcanoPlot.R' - 'customConverters.R' - 'datasets.R' - 'retrieveBiomart.R' - 'samr-adapted.R' + 'DIscBIO-classes.R' + 'DIscBIO-generic-ClassVectoringDT.R' + 'DIscBIO-generic-Clustexp.R' + 'DIscBIO-generic-DEGanalysis.R' + 'DIscBIO-generic-DEGanalysis2clust.R' + 'DIscBIO-generic-Exprmclust.R' + 'DIscBIO-generic-FinalPreprocessing.R' + 'DIscBIO-generic-FindOutliersKM.R' + 'DIscBIO-generic-FindOutliersMB.R' + 'DIscBIO-generic-KMClustDiffGenes.R' + 'DIscBIO-generic-KMclustheatmap.R' + 'DIscBIO-generic-KmeanOrder.R' + 'DIscBIO-generic-MBClustDiffGenes.R' + 'DIscBIO-generic-MBclustheatmap.R' + 'DIscBIO-generic-NoiseFiltering.R' + 'DIscBIO-generic-Normalizedata.R' + 'DIscBIO-generic-PCAplotSymbols.R' + 'DIscBIO-generic-PlotmclustMB.R' + 'DIscBIO-generic-comptSNE.R' + 'DIscBIO-generic-comptsneMB.R' + 'DIscBIO-generic-plotExptSNE.R' + 'DIscBIO-generic-plotGap.R' + 'DIscBIO-generic-plotKmeansLabelstSNE.R' + 'DIscBIO-generic-plotMBLabelstSNE.R' + 'DIscBIO-generic-plotOrderKMtsne.R' + 'DIscBIO-generic-plotOrderMBtsne.R' + 'DIscBIO-generic-plotSilhouette.R' + 'DIscBIO-generic-plotSymbolstSNE.R' + 'DIscBIO-generic-plotexptsneMB.R' + 'DIscBIO-generic-plotsilhouetteMB.R' + 'DIscBIO-generic-plottSNE.R' + 'DIscBIO-generic-plottsneMB.R' + 'J48DT.R' + 'J48DTeval.R' + 'Jaccard.R' + 'MB_Order.R' + 'NetAnalysis.R' + 'Networking.R' + 'PPI.R' + 'PlotMBexpPCA.R' + 'PlotMBorderPCA.R' + 'RpartDT.R' + 'RpartEVAL.R' + 'VolcanoPlot.R' + 'customConverters.R' + 'datasets.R' + 'reformatSiggenes.R' + 'replaceDecimals.R' + 'retrieveBiomart.R' + 'samr-adapted.R' diff --git a/R/reformatSiggenes.R b/R/reformatSiggenes.R index 6d60a28..49e28fd 100644 --- a/R/reformatSiggenes.R +++ b/R/reformatSiggenes.R @@ -1,5 +1,6 @@ #' @title Reformat Siggenes Table #' @description Reformats the Siggenes table output from the SAMR package +#' @param table output from `samr::samr.compute.siggenes.table` #' @seealso replaceDecimals reformatSiggenes <- function(table) { if (is.null(table)) return(table) diff --git a/R/replaceDecimals.R b/R/replaceDecimals.R index dd50bd2..6da146a 100644 --- a/R/replaceDecimals.R +++ b/R/replaceDecimals.R @@ -1,6 +1,10 @@ #' @title Replace Decimals -#' @description Replaces decimals separators between comma and periods +#' @description Replaces decimals separators between comma and periods on a +#' character vector #' @note This function was especially designed to be used with retormatSiggenes +#' @param x vector of characters +#' @param from decimal separator on input file +#' @param to decimal separator for output file #' @seealso reformatSiggenes replaceDecimals <- function(x, from=",", to=".") { x <- gsub(",", ".", x) diff --git a/man/VolcanoPlot.Rd b/man/VolcanoPlot.Rd index 09b48f1..f733247 100644 --- a/man/VolcanoPlot.Rd +++ b/man/VolcanoPlot.Rd @@ -4,7 +4,7 @@ \alias{VolcanoPlot} \title{Volcano Plot} \usage{ -VolcanoPlot(object, value = 0.05, name, fc = 0.5, FS = 0.4) +VolcanoPlot(object, value = 0.05, name = NULL, fc = 0.5, FS = 0.4) } \arguments{ \item{object}{A data frame showing the differentially expressed genes (DEGs) diff --git a/man/reformatSiggenes.Rd b/man/reformatSiggenes.Rd index 8202e81..279842e 100644 --- a/man/reformatSiggenes.Rd +++ b/man/reformatSiggenes.Rd @@ -6,6 +6,9 @@ \usage{ reformatSiggenes(table) } +\arguments{ +\item{table}{output from `samr::samr.compute.siggenes.table`} +} \description{ Reformats the Siggenes table output from the SAMR package } diff --git a/man/replaceDecimals.Rd b/man/replaceDecimals.Rd index 074ce27..6e914bf 100644 --- a/man/replaceDecimals.Rd +++ b/man/replaceDecimals.Rd @@ -6,8 +6,16 @@ \usage{ replaceDecimals(x, from = ",", to = ".") } +\arguments{ +\item{x}{vector of characters} + +\item{from}{decimal separator on input file} + +\item{to}{decimal separator for output file} +} \description{ -Replaces decimals separators between comma and periods +Replaces decimals separators between comma and periods on a +character vector } \note{ This function was especially designed to be used with retormatSiggenes From 5e8cc5ee3eea5a4c7c550be8974865d1c671915a Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 29 Jun 2020 16:16:43 +0200 Subject: [PATCH 23/99] Fixed unit tests --- tests/testthat/test.DIscBIO.IMP.R | 8 ++++---- 1 file changed, 4 insertions(+), 4 deletions(-) diff --git a/tests/testthat/test.DIscBIO.IMP.R b/tests/testthat/test.DIscBIO.IMP.R index 2b128e7..b5e3925 100644 --- a/tests/testthat/test.DIscBIO.IMP.R +++ b/tests/testthat/test.DIscBIO.IMP.R @@ -123,9 +123,9 @@ test_that("DEGs are calculated", { object = sapply(cdiff2, function(x) class(x)[1]), expected = c("matrix", "data.frame") ) - expect_identical( + expect_equivalent( object = sapply(cdiff3, function(x) class(x)[1]), - expected = c("matrix", "data.frame") + expected = c("matrix", "data.frame", "data.frame", "data.frame") ) }) @@ -249,9 +249,9 @@ test_that("DEGs are calculated", { object = sapply(cdiff2, function(x) class(x)[1]), expected = c("matrix", "data.frame") ) - expect_identical( + expect_equivalent( object = sapply(cdiff3, function(x) class(x)[1]), - expected = c("matrix", "data.frame") + expected = c("matrix", "data.frame", "data.frame", "data.frame") ) }) From 1dcd0f431be882efb42969128d0dd4000d3dad2f Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Wed, 1 Jul 2020 14:16:21 +0200 Subject: [PATCH 24/99] Removed code execution on vignette --- vignettes/DIscBIO-vignette.Rmd | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/vignettes/DIscBIO-vignette.Rmd b/vignettes/DIscBIO-vignette.Rmd index e6d346f..06bdba8 100644 --- a/vignettes/DIscBIO-vignette.Rmd +++ b/vignettes/DIscBIO-vignette.Rmd @@ -16,7 +16,7 @@ The pipeline consists of four successive steps: data pre-processing, cellular cl ## Required Packages ```{r options, echo=FALSE} -knitr::opts_chunk$set(fig.width=7, fig.height=7) +knitr::opts_chunk$set(fig.width=7, fig.height=7, eval=FALSE) ``` ```{r} From 519af8eebb31f26cd98572015a9e662c2d3fcb2d Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Wed, 1 Jul 2020 14:30:53 +0200 Subject: [PATCH 25/99] Fixed exporting example --- R/DIscBIO-generic-MBClustDiffGenes.R | 2 +- man/MBClustDiffGenes.Rd | 2 +- 2 files changed, 2 insertions(+), 2 deletions(-) diff --git a/R/DIscBIO-generic-MBClustDiffGenes.R b/R/DIscBIO-generic-MBClustDiffGenes.R index 9c06e05..317bd9c 100644 --- a/R/DIscBIO-generic-MBClustDiffGenes.R +++ b/R/DIscBIO-generic-MBClustDiffGenes.R @@ -25,7 +25,7 @@ #' sc <- comptsneMB(sc, max_iter=100) #' sc <- Clustexp(sc, cln=3) #' sc <- MB_Order(sc, export = FALSE) -#' cdiff <- MBClustDiffGenes(sc, K=3, fdr=.1) +#' cdiff <- MBClustDiffGenes(sc, K=3, fdr=.1, export=FALSE) #' str(cdiff) #' } setGeneric( diff --git a/man/MBClustDiffGenes.Rd b/man/MBClustDiffGenes.Rd index 4175937..910f567 100644 --- a/man/MBClustDiffGenes.Rd +++ b/man/MBClustDiffGenes.Rd @@ -56,7 +56,7 @@ sc <- Exprmclust(sc, K=3) sc <- comptsneMB(sc, max_iter=100) sc <- Clustexp(sc, cln=3) sc <- MB_Order(sc, export = FALSE) -cdiff <- MBClustDiffGenes(sc, K=3, fdr=.1) +cdiff <- MBClustDiffGenes(sc, K=3, fdr=.1, export=FALSE) str(cdiff) } } From 04131d54f55137c9c77fc096a6c889f5e9c27c69 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Thu, 2 Jul 2020 08:27:13 +0200 Subject: [PATCH 26/99] Added prepped dataset --- DESCRIPTION | 1 + R/prepExampleDataset.R | 36 ++++++++++++++++++++++++++ data/valuesG1msReduced_treated_K.rda | Bin 0 -> 393192 bytes data/valuesG1msReduced_treated_MB.rda | Bin 0 -> 378562 bytes man/prepExampleDataset.Rd | 18 +++++++++++++ 5 files changed, 55 insertions(+) create mode 100644 R/prepExampleDataset.R create mode 100644 data/valuesG1msReduced_treated_K.rda create mode 100644 data/valuesG1msReduced_treated_MB.rda create mode 100644 man/prepExampleDataset.Rd diff --git a/DESCRIPTION b/DESCRIPTION index e875085..373e866 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -101,6 +101,7 @@ Collate: 'VolcanoPlot.R' 'customConverters.R' 'datasets.R' + 'prepExampleDataset.R' 'reformatSiggenes.R' 'replaceDecimals.R' 'retrieveBiomart.R' diff --git a/R/prepExampleDataset.R b/R/prepExampleDataset.R new file mode 100644 index 0000000..c53cacd --- /dev/null +++ b/R/prepExampleDataset.R @@ -0,0 +1,36 @@ +#' @title Prepare Example Dataset +#' @description Internal function that prepares a pre-treated dataset for use in +#' several examples +#' @param clustering determines the clustering method (either "K" or "MB"). +#' @note This is an internal function, reachable through the terminal by using `DIscBIO:::prepExampleDataset` +#' @return rda file +prepExampleDataset <- function(clustering) { + # ========================================================================== + # Initial data treatment + # ========================================================================== + message("Treating dataset") + sc <- DISCBIO(valuesG1msReduced) + sc <- NoiseFiltering( + sc, percentile=0.9, CV=0.2, export=FALSE, plot=FALSE, quiet=TRUE + ) + sc <- Normalizedata(sc) + sc <- FinalPreprocessing(sc, export=FALSE, quiet=TRUE) + # ========================================================================== + # Clustering + # ========================================================================== + message("Clustering") + if (clustering == "K") { + sc <- Clustexp(sc, cln=3, quiet=TRUE) + sc <- comptSNE(sc, quiet=TRUE) + output_name <- "valuesG1msReduced_treated_K" + } else if (clustering == "MB") { + sc <- Exprmclust(sc, quiet=TRUE) + sc <- comptsneMB(sc, rseed=15555, quiet=TRUE) + output_name <- "valuesG1msReduced_treated_MB" + } + # ========================================================================== + # Output + # ========================================================================== + message("Saving") + save(sc, file = paste0("data/", output_name, ".rda")) +} \ No newline at end of file diff --git a/data/valuesG1msReduced_treated_K.rda b/data/valuesG1msReduced_treated_K.rda new file mode 100644 index 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zrgxn{Az5=b$c&o%tJo3&Qen^^$HGK!O70ASoqa-OIiTR^#+}t#_vmFr_YjmFRHTl4%_@^+;Y_j%*u(b(G9K#)E=la zV)Cc8${0rbDN8oQcH31Q|IC_6!=33!`-WCLfDG5{ai;nL3?A*<^aeG~lf}GP6uIyp zT23V*H-O!Jtu}G4E3#A?{s&RXFRo`%TJ6(kcDQ|Ls$2Z9V{N4>`{6&2H=Pd>XQFY; zceCgvf&!m6<1dsgOFhPsixiTOS=Z1)8HnsCG_%Pf92SuMuK-uIRD+V`fm!$@QogX1-Vi<{fP zOvvmozC_}im&72Q$8$s(X8?kY>UwnG=(^2L~btG0xX#h?bkOW?g_sH)L8IXu~@s(X>d_J0U}i=lu1A&b8f@BU%?w;1}5{QpM$EBG(s zzpO~$zxbt*L&%3_j%99o`SG8E5NT3O?3@YGx(`z3(Xp{}f&aNu`j5!}vG;#lr(^bi z8u>3U676r332zXobDRLrT-qpHr%m1BR+rekUaV+B;X1R~ZnhyE%+LA5ol^M~TjzA4 M8)alGudtu|KWB*qLI3~& literal 0 HcmV?d00001 diff --git a/man/prepExampleDataset.Rd b/man/prepExampleDataset.Rd new file mode 100644 index 0000000..71d3b61 --- /dev/null +++ b/man/prepExampleDataset.Rd @@ -0,0 +1,18 @@ +% Generated by roxygen2: do not edit by hand +% Please edit documentation in R/prepExampleDataset.R +\name{prepExampleDataset} +\alias{prepExampleDataset} +\title{Prepare Example Dataset} +\usage{ +prepExampleDataset(clustering) +} +\arguments{ +\item{clustering}{determines the clustering method (either "K" or "MB").} +} +\value{ +rda file +} +\description{ +Internal function that prepares a pre-treated dataset for use in +several examples +} From 27197ebf139af13907a20d44b1ce5432806a3332 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Thu, 2 Jul 2020 08:46:58 +0200 Subject: [PATCH 27/99] Improved treated datasets --- R/datasets.R | 14 ++++++++ R/prepExampleDataset.R | 45 +++++++++++++++++--------- data/valuesG1msReduced_treated_K.rda | Bin 393192 -> 393211 bytes data/valuesG1msReduced_treated_MB.rda | Bin 378562 -> 378583 bytes man/prepExampleDataset.Rd | 11 +++++-- man/valuesG1msReduced_treated_K.Rd | 9 ++++++ man/valuesG1msReduced_treated_MB.Rd | 9 ++++++ 7 files changed, 70 insertions(+), 18 deletions(-) create mode 100644 man/valuesG1msReduced_treated_K.Rd create mode 100644 man/valuesG1msReduced_treated_MB.Rd diff --git a/R/datasets.R b/R/datasets.R index 0424018..87ffd2f 100644 --- a/R/datasets.R +++ b/R/datasets.R @@ -25,3 +25,17 @@ NULL #' @name HumanMouseGeneIds #' @docType data NULL + +#' @title Single-cells data from a myxoid liposarcoma cell line (K variant) +#' +#' @description Equivalent to valuesG1msReduced after treatment and K-means clustering. +#' @name valuesG1msReduced_treated_K +#' @docType data +NULL + +#' @title Single-cells data from a myxoid liposarcoma cell line (MB variant) +#' +#' @description Equivalent to valuesG1msReduced after treatment and model-based clustering. +#' @name valuesG1msReduced_treated_MB +#' @docType data +NULL \ No newline at end of file diff --git a/R/prepExampleDataset.R b/R/prepExampleDataset.R index c53cacd..5641e9e 100644 --- a/R/prepExampleDataset.R +++ b/R/prepExampleDataset.R @@ -1,15 +1,19 @@ #' @title Prepare Example Dataset #' @description Internal function that prepares a pre-treated dataset for use in #' several examples -#' @param clustering determines the clustering method (either "K" or "MB"). -#' @note This is an internal function, reachable through the terminal by using `DIscBIO:::prepExampleDataset` +#' @param dataset Dataset used for transformation +#' @param save save results? +#' @examples +#' \donttest{ +#' DIscBIO:::prepExampleDataset(valuesG1msReduced, save=FALSE) +#' } #' @return rda file -prepExampleDataset <- function(clustering) { +prepExampleDataset <- function(dataset, save=TRUE) { # ========================================================================== # Initial data treatment # ========================================================================== message("Treating dataset") - sc <- DISCBIO(valuesG1msReduced) + sc <- DISCBIO(dataset) sc <- NoiseFiltering( sc, percentile=0.9, CV=0.2, export=FALSE, plot=FALSE, quiet=TRUE ) @@ -18,19 +22,28 @@ prepExampleDataset <- function(clustering) { # ========================================================================== # Clustering # ========================================================================== - message("Clustering") - if (clustering == "K") { - sc <- Clustexp(sc, cln=3, quiet=TRUE) - sc <- comptSNE(sc, quiet=TRUE) - output_name <- "valuesG1msReduced_treated_K" - } else if (clustering == "MB") { - sc <- Exprmclust(sc, quiet=TRUE) - sc <- comptsneMB(sc, rseed=15555, quiet=TRUE) - output_name <- "valuesG1msReduced_treated_MB" - } + message("K-means clustering") + sc_k <- Clustexp(sc, cln=3, quiet=TRUE) + sc_k <- comptSNE(sc_k, quiet=TRUE) + valuesG1msReduced_treated_K <- sc_k + message("Model-based clustering") + sc_mb <- Exprmclust(sc, quiet=TRUE) + sc_mb <- comptsneMB(sc_mb, rseed=15555, quiet=TRUE) + valuesG1msReduced_treated_MB <- sc_mb # ========================================================================== # Output # ========================================================================== - message("Saving") - save(sc, file = paste0("data/", output_name, ".rda")) + message("Saving datasets") + if (save) { + save( + valuesG1msReduced_treated_K, + file = "data/valuesG1msReduced_treated_K.rda" + ) + save( + valuesG1msReduced_treated_MB, + file = "data/valuesG1msReduced_treated_MB.rda" + ) + } else { + message("Not saving dataset because (save == FALSE)") + } } \ No newline at end of file diff --git a/data/valuesG1msReduced_treated_K.rda b/data/valuesG1msReduced_treated_K.rda index 040b0ae45ee0b374634cce168f9ab7937a69ba0d..0daf66ba254beab231335d60d7b5d1d8c99ae75c 100644 GIT binary patch delta 365330 zcmaHRi96Jb_kZP@*Xwm&t3qj>P&x?mtqXa+rl?f)LS zbL8B!JBR;%@FVH9qug3{uGioHdl9rJ^XXyApsvJyvzSAFB^MmuZ*~{G(aN-4p+szk zpZFUsBno-LYIr7uwHYx0Yc8SNxMEO+S(sas=y_)9WALtdcDOhFTIuxCKt04S=P@oF zoVHhCEO4C5Rl*380CNI%bT+P<7PO861@siLplhG2O`se(PDJGQ@Qe+6H04NPHc~ij z+B#8Gh(X6jgKq7X3=iDe?F)Rh5t0+KtgV#ozpkxBW-SJqm=wY8@jis(S8poIC*w9& zs2tjFzIWkgG=0LoH+q_0Ue}{2`8l#j#1tzmQiAS`R09Jr>TUP%-AqYs{qTT?=Gni6 zWt%XpU zyw}VGhMiA0wN5>SwlEYWK6_C(~D-@bGQTN_a{!j|4C&~oJ`Taix$CK3VtLN*k?x0Qd6&CeB;o}-s zoZUk4ro(RLjny84cNhUAu%?*gaaCD}y9UH8DQFXEw^VxbvoIlV8FT3=rh0}y2@^u{ 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z3%S&uK?r~`2n(QKb8gLgTYo4zp{Dva$?tpAX$-W%PT zR+Vo%w_E!<2@boL7bz)L&hFmur1Pf4=-A*Z$?${l^8Igkce;y{`b$ahig*?P;OI5E z@UTMS*+W+V^swcu99^aeNmjnVF`5o>FsA7qxH`g8ypw{~1;Pzv^FNpWs!(|!qe!QZ zbMp3}(-$@%q;cQe2LW@r3jO95F&d{dTvQPs&HI-kMP+9g47P7B)K^@U;bnre{)P%D z8-!beCSF(CG@SU}yE|HYM9Bl%1B@mR?`$0+nnD%cIWC1IHwIxg5pNWGKQ29<$t3ls z;c46mun}N14xg1PV=OAUpEdhihMGeafF^vkLTHCtG?qA{4A8{ai4Wp<^7}tGH~yA< zKjb4hpk!no7LKp~YiHy?7tj08fc?`*z7Bt>-nf$z_h$u6=jVO)?)$cH-REcNyT=dT z2vcKI<3+}%i;T@(O)M-;ms*-o4UA1Jjg9>WBa{By1+3uhO#krz{el_4SDBL5NwZ&X Km%D1q!T$n!0`(C9 From 5c51b99254562763b58bf1ca07f87e0f7a445961 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Wed, 29 Jul 2020 17:37:27 +0200 Subject: [PATCH 64/99] Fixed function syntax --- R/samr-adapted.R | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/R/samr-adapted.R b/R/samr-adapted.R index 98864d8..a4aca5a 100644 --- a/R/samr-adapted.R +++ b/R/samr-adapted.R @@ -1151,7 +1151,7 @@ foldchange.seq.twoclass.unpaired <- function(x, y, depth) { x.norm <- scale(x, center = F, scale = depth) + 1e-08 fc <- apply(x.norm[, y == 2], 1, median) / - apply(x.norm[, y == 1, 1, median]) + apply(x.norm[, y == 1], 1, median) return(fc) } integer.base.b <- function(x, b = 2) { From 7fa1ae473ef6c4e19b1477c01dc03c8f8ef38e01 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Wed, 29 Jul 2020 17:37:50 +0200 Subject: [PATCH 65/99] Added URL to GitHub page (vignette) --- DESCRIPTION | 1 + 1 file changed, 1 insertion(+) diff --git a/DESCRIPTION b/DESCRIPTION index fbd1f7b..29de301 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -53,6 +53,7 @@ Suggests: VignetteBuilder: knitr LazyData: true RoxygenNote: 7.1.1 +URL: https://github.com/ocbe-uio/DIscBIO#usage BugReports: https://github.com/ocbe-uio/DIscBIO/issues Collate: 'DIscBIO-classes.R' From ed165f31ea203a5b1a7d54a00dd6656360ef0d76 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Thu, 30 Jul 2020 10:27:01 +0200 Subject: [PATCH 66/99] Removed leftover vignette builder --- DESCRIPTION | 3 +-- 1 file changed, 1 insertion(+), 2 deletions(-) diff --git a/DESCRIPTION b/DESCRIPTION index 29de301..b700d57 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -1,5 +1,5 @@ Package: DIscBIO -Date: 2020-07-29 +Date: 2020-07-30 Title: A User-Friendly Pipeline for Biomarker Discovery in Single-Cell Transcriptomics Version: 1.0.0 Authors@R: @@ -50,7 +50,6 @@ Suggests: enrichR, Seurat, BiocManager -VignetteBuilder: knitr LazyData: true RoxygenNote: 7.1.1 URL: https://github.com/ocbe-uio/DIscBIO#usage From 7c2f508094aa5501f4bfa964517c1912139c918d Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 3 Aug 2020 09:46:36 +0200 Subject: [PATCH 67/99] Removed integration tests --- tests/testthat/test-integration.R | 22 ---------------------- 1 file changed, 22 deletions(-) delete mode 100644 tests/testthat/test-integration.R diff --git a/tests/testthat/test-integration.R b/tests/testthat/test-integration.R deleted file mode 100644 index cad4f06..0000000 --- a/tests/testthat/test-integration.R +++ /dev/null @@ -1,22 +0,0 @@ -# ============================================================================== -# Testing integration with SingleCellExperiment -# ============================================================================== -context("Converting other formats to DISCBIO") -# ------------------------------------------------------------------------------ -# Setting up datasets -# ------------------------------------------------------------------------------ -pmbc_seurat <- Seurat::pbmc_small -pmbc_sce <- Seurat::as.SingleCellExperiment(pmbc_seurat) -g1_sce <- SingleCellExperiment::SingleCellExperiment( - list(counts=as.matrix(valuesG1msRed)) -) -# ------------------------------------------------------------------------------ -# Performing unit tests -# ------------------------------------------------------------------------------ -test_that("Pure text gets formatted as DISCBIO", { - expect_s4_class(as.DISCBIO(pmbc_seurat), "DISCBIO") -}) -test_that("SCE file gets formatted as DISCBIO", { - expect_s4_class(as.DISCBIO(pmbc_sce), "DISCBIO") - expect_s4_class(as.DISCBIO(g1_sce), "DISCBIO") -}) From 738d9d51339b35cc4e0264a39876853701eaf39d Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 3 Aug 2020 09:46:44 +0200 Subject: [PATCH 68/99] Further test simplification --- tests/testthat/test-DIscBIO.IMP.R | 31 +++---------------------------- 1 file changed, 3 insertions(+), 28 deletions(-) diff --git a/tests/testthat/test-DIscBIO.IMP.R b/tests/testthat/test-DIscBIO.IMP.R index bb5a492..e6e838e 100644 --- a/tests/testthat/test-DIscBIO.IMP.R +++ b/tests/testthat/test-DIscBIO.IMP.R @@ -46,8 +46,8 @@ test_that("Data is normalized", { context("K-means clustering") -sc <- Clustexp(sc, cln=2, quiet=TRUE, rseed=17000) # K-means clustering -sc <- comptSNE(sc, rseed=15555, quiet=TRUE, max_iter=10) +sc <- Clustexp(sc, clustnr=2, cln=2, bootnr=10, quiet=TRUE, rseed=17000) +sc <- comptSNE(sc, rseed=15555, quiet=TRUE, max_iter=1, epoch=10) test_that("tSNE is computed", { expect_equal(class(sc@tsne), "data.frame") @@ -95,7 +95,7 @@ context("Differential Expression Analysis") cdiff3 <- suppressMessages( DEGanalysis2clust( sc, Clustering="K-means", K=2, fdr=.2, name="Name", First="CL1", - Second="CL2", export=FALSE, quiet=TRUE, plot=FALSE, nperms=5, nresamp=2 + Second="CL2", export=FALSE, quiet=TRUE, plot=FALSE, nperms=2, nresamp=2 ) ) @@ -105,28 +105,3 @@ test_that("DEGs are calculated", { expected = c("matrix", "data.frame", "data.frame", "data.frame") ) }) - -# Decision tree -sigDEG <- cdiff3[[1]] - -DATAforDT <- ClassVectoringDT( - sc, Clustering="K-means", K=2, First="CL1", Second="CL2", sigDEG, - quiet = TRUE -) - -j48dt <- J48DT(DATAforDT, quiet = TRUE, plot = FALSE) -j48dt_eval <- J48DTeval( - DATAforDT, num.folds=10, First="CL1", Second="CL2", quiet=TRUE -) -rpartEVAL <- RpartEVAL( - DATAforDT, num.folds=10, First="CL1", Second="CL2", quiet = TRUE -) - -test_that("Decision tree elements are defined", { - expect_output(str(DATAforDT), "5 obs. of 21 variables") - expect_identical( - attributes(j48dt)$class, c("J48", "Weka_tree", "Weka_classifier") - ) - expect_identical(j48dt_eval, c(TP = 7, FN = 4, FP = 5, TN = 5)) - expect_identical(rpartEVAL, c(TP = 5, FN = 6, FP = 10, TN = 0)) -}) From 09b4867dcc7a3321d3a5292d4973fa80abf0bb58 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 3 Aug 2020 11:17:50 +0200 Subject: [PATCH 69/99] Merged plotSilhouette and plotSilhouetteMB --- DESCRIPTION | 1 - NAMESPACE | 1 - R/DIscBIO-generic-plotSilhouette.R | 33 ++++++++++++++++++---------- R/DIscBIO-generic-plotsilhouetteMB.R | 33 ---------------------------- man/plotsilhouetteMB.Rd | 28 ----------------------- 5 files changed, 22 insertions(+), 74 deletions(-) delete mode 100644 R/DIscBIO-generic-plotsilhouetteMB.R delete mode 100644 man/plotsilhouetteMB.Rd diff --git a/DESCRIPTION b/DESCRIPTION index b700d57..10fa716 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -84,7 +84,6 @@ Collate: 'DIscBIO-generic-plotSilhouette.R' 'DIscBIO-generic-plotSymbolstSNE.R' 'DIscBIO-generic-plotexptsneMB.R' - 'DIscBIO-generic-plotsilhouetteMB.R' 'DIscBIO-generic-plottSNE.R' 'DIscBIO-generic-plottsneMB.R' 'J48DT.R' diff --git a/NAMESPACE b/NAMESPACE index 5c00ab0..a87392d 100644 --- a/NAMESPACE +++ b/NAMESPACE @@ -47,7 +47,6 @@ exportMethods(plotOrderMBtsne) exportMethods(plotSilhouette) exportMethods(plotSymbolstSNE) exportMethods(plotexptsneMB) -exportMethods(plotsilhouetteMB) exportMethods(plottSNE) exportMethods(plottsneMB) import(org.Hs.eg.db) diff --git a/R/DIscBIO-generic-plotSilhouette.R b/R/DIscBIO-generic-plotSilhouette.R index 8d6c1f5..3986d5b 100644 --- a/R/DIscBIO-generic-plotSilhouette.R +++ b/R/DIscBIO-generic-plotSilhouette.R @@ -22,16 +22,27 @@ setMethod( f = "plotSilhouette", signature = "DISCBIO", definition = function(object, K) { - if (length(object@kmeans$kpart) == 0) { - stop("run clustexp before plotsilhouette") - } - if (length(unique(object@kmeans$kpart)) < 2) { - stop("only a single cluster: no silhouette plot") - } - col <- c("black", "blue", "green", "red", "yellow", "gray") - kpart <- object@kmeans$kpart - distances <- dist.gen(object@distances) - si <- silhouette(kpart, distances) - plot(si, col = col[1:K]) + # ====================================================================== + # Validation + # ====================================================================== + ran_clustexp <- length(object@kmeans$kpart) > 0 + ran_exprmclust <- length(object@MBclusters$clusterid) > 0 + if (ran_clustexp) { + kpart <- object@kmeans$kpart + } else if (ran_exprmclust) { + kpart <- object@MBclusters$clusterid + } else { + stop("run clustexp or exprmclust before plotSilhouette") + } + if (length(unique(kpart)) < 2) { + stop("only a single cluster: no silhouette plot") + } + # ====================================================================== + # Plotting + # ====================================================================== + col <- c("black", "blue", "green", "red", "yellow", "gray") + distances <- dist.gen(object@distances) + si <- silhouette(kpart, distances) + plot(si, col = col[1:K]) } ) \ No newline at end of file diff --git a/R/DIscBIO-generic-plotsilhouetteMB.R b/R/DIscBIO-generic-plotsilhouetteMB.R deleted file mode 100644 index 814aae6..0000000 --- a/R/DIscBIO-generic-plotsilhouetteMB.R +++ /dev/null @@ -1,33 +0,0 @@ -#' @title Silhouette Plot for Model-based clustering -#' @description The silhouette provides a representation of how well each point -#' is represented by its cluster in comparison to the closest neighboring -#' cluster. It computes for each point the difference between the average -#' similarity to all points in the same cluster and to all points in the -#' closest neighboring cluster. This difference it normalize such that it can -#' take values between -1 and 1 with higher values reflecting better -#' representation of a point by its cluster. -#' @param object \code{DISCBIO} class object. -#' @param K A numeric value of the number of clusters -#' @importFrom stats as.dist cor -#' @importFrom cluster silhouette -#' @return A silhouette plot -setGeneric("plotsilhouetteMB", function(object, K) - standardGeneric("plotsilhouetteMB")) - -#' @export -#' @rdname plotsilhouetteMB -setMethod( - "plotsilhouetteMB", - signature = "DISCBIO", - definition = function(object, K) { - if (length(object@MBclusters$clusterid) == 0) - stop("run exprmclust before plotsilhouetteMB") - if (length(unique(object@MBclusters$clusterid)) < 2) - stop("only a single cluster: no silhouette plot") - col = c("black", "blue", "green", "red", "yellow", "gray") - kpart <- object@MBclusters$clusterid - distances <- dist.gen(t(object@fdata)) - si <- silhouette(kpart, distances) - plot(si, col = col[1:K]) - } -) \ No newline at end of file diff --git a/man/plotsilhouetteMB.Rd b/man/plotsilhouetteMB.Rd deleted file mode 100644 index 55ec02c..0000000 --- a/man/plotsilhouetteMB.Rd +++ /dev/null @@ -1,28 +0,0 @@ -% Generated by roxygen2: do not edit by hand -% Please edit documentation in R/DIscBIO-generic-plotsilhouetteMB.R -\name{plotsilhouetteMB} -\alias{plotsilhouetteMB} -\alias{plotsilhouetteMB,DISCBIO-method} -\title{Silhouette Plot for Model-based clustering} -\usage{ -plotsilhouetteMB(object, K) - -\S4method{plotsilhouetteMB}{DISCBIO}(object, K) -} -\arguments{ -\item{object}{\code{DISCBIO} class object.} - -\item{K}{A numeric value of the number of clusters} -} -\value{ -A silhouette plot -} -\description{ -The silhouette provides a representation of how well each point - is represented by its cluster in comparison to the closest neighboring - cluster. It computes for each point the difference between the average - similarity to all points in the same cluster and to all points in the - closest neighboring cluster. This difference it normalize such that it can - take values between -1 and 1 with higher values reflecting better - representation of a point by its cluster. -} From 14ad04bff73bca331039197a6f3c740336e29b35 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 3 Aug 2020 11:40:59 +0200 Subject: [PATCH 70/99] Merged PlotMBorderPCA and PlotMBexpPCA --- DESCRIPTION | 3 +- NAMESPACE | 3 +- R/PlotMBexpPCA.R | 82 ---------------------------------- R/PlotMBorderPCA.R | 67 ---------------------------- R/PlotMBpca.R | 100 ++++++++++++++++++++++++++++++++++++++++++ man/PlotMBexpPCA.Rd | 25 ----------- man/PlotMBorderPCA.Rd | 18 -------- man/PlotMBpca.Rd | 27 ++++++++++++ 8 files changed, 129 insertions(+), 196 deletions(-) delete mode 100644 R/PlotMBexpPCA.R delete mode 100644 R/PlotMBorderPCA.R create mode 100644 R/PlotMBpca.R delete mode 100644 man/PlotMBexpPCA.Rd delete mode 100644 man/PlotMBorderPCA.Rd create mode 100644 man/PlotMBpca.Rd diff --git a/DESCRIPTION b/DESCRIPTION index 10fa716..ed73306 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -93,8 +93,7 @@ Collate: 'NetAnalysis.R' 'Networking.R' 'PPI.R' - 'PlotMBexpPCA.R' - 'PlotMBorderPCA.R' + 'PlotMBpca.R' 'RpartDT.R' 'RpartEVAL.R' 'VolcanoPlot.R' diff --git a/NAMESPACE b/NAMESPACE index a87392d..f319841 100644 --- a/NAMESPACE +++ b/NAMESPACE @@ -11,8 +11,7 @@ export(MB_Order) export(NetAnalysis) export(Networking) export(PPI) -export(PlotMBexpPCA) -export(PlotMBorderPCA) +export(PlotMBpca) export(RpartDT) export(RpartEVAL) export(VolcanoPlot) diff --git a/R/PlotMBexpPCA.R b/R/PlotMBexpPCA.R deleted file mode 100644 index 55eb822..0000000 --- a/R/PlotMBexpPCA.R +++ /dev/null @@ -1,82 +0,0 @@ -#' @title Plotting gene expression in Model-based clustering in PCA. -#' @description The PCA representation can also be used to show the gene -#' expression of a particular gene. -#' @param object \code{DISCBIO} class object. -#' @param g Individual gene name or vector with a group of gene names -#' corresponding to a subset of valid row names of the \code{ndata} slot of -#' the \code{DISCBIO} object. -#' @param n String of characters representing the title of the plot. Default is -#' NULL and the first element of \code{g} is chosen. -#' @importFrom RColorBrewer brewer.pal -#' @importFrom grDevices colorRampPalette -#' @importFrom graphics layout par image -#' @return A plot of the PCA. -#' @export - -PlotMBexpPCA <- function(object, g, n = NULL) { - if (length(intersect(g, rownames(object@ndata))) < length(unique(g))) - stop( - "second argument does not correspond to set of rownames slot", - "ndata of SCseq object" - ) - if (is.null(n)) - n <- g[1] - data = object@MBclusters - #Expression<-cbind(data$pcareduceres,object@ndata[g,]) - l <- apply(object@ndata[g,] - .1, 2, sum) + .1 - #l <- Expression[,3] - mi <- min(l, na.rm = TRUE) - ma <- max(l, na.rm = TRUE) - ColorRamp <- - colorRampPalette(rev(brewer.pal(n = 11, name = "RdYlBu")))(100) - ColorLevels <- seq(mi, ma, length = length(ColorRamp)) - v <- round((l - mi) / (ma - mi) * 99 + 1, 0) - layout( - matrix( - data = c(1, 3, 2, 4), - nrow = 2, - ncol = 2 - ), - widths = c(5, 1, 5, 1), - heights = c(5, 1, 1, 1) - ) - opar <- par(mar = c(5, 5, 2.5, 2)) - on.exit(par(opar)) - plot( - data$pcareduceres[, 1], - data$pcareduceres[, 2], - xlab = "PC1", - ylab = "PC2", - pch = 20, - cex = 0, - col = "grey", - las = 1, - main = n - ) - for (k in 1:length(v)) { - points( - data$pcareduceres[k, 1], - data$pcareduceres[k, 2], - col = ColorRamp[v[k]], - pch = 20, - cex = 2 - ) - } - opar <- par(mar = c(3, 2.5, 2.5, 2)) - on.exit(par(opar)) - image( - 1, - ColorLevels, - matrix( - data = ColorLevels, - ncol = length(ColorLevels), - nrow = 1 - ), - col = ColorRamp, - xlab = "", - ylab = "", - las = 2, - xaxt = "n" - ) - layout(1) -} \ No newline at end of file diff --git a/R/PlotMBorderPCA.R b/R/PlotMBorderPCA.R deleted file mode 100644 index 2d618f1..0000000 --- a/R/PlotMBorderPCA.R +++ /dev/null @@ -1,67 +0,0 @@ -#' @title Plotting pseudo-time ordering in Model-based clustering in PCA. -#' @description The PCA representation can also be used to show the pseudo-time -#' ordering. -#' @param object \code{DISCBIO} class object. -#' @importFrom RColorBrewer brewer.pal -#' @importFrom grDevices colorRampPalette -#' @importFrom graphics layout par image -#' @export -#' @return A plot of the PCA. -PlotMBorderPCA <- function(object) { - data = object@MBclusters - MBordertable <- cbind(data$pcareduceres, object@MBordering) - l <- MBordertable[, 3] - mi <- min(l, na.rm = TRUE) - ma <- max(l, na.rm = TRUE) - ColorRamp <- - colorRampPalette(rev(brewer.pal(n = 11, name = "RdYlBu")))(100) - ColorLevels <- seq(mi, ma, length = length(ColorRamp)) - v <- round((l - mi) / (ma - mi) * 99 + 1, 0) - layout( - matrix( - data = c(1, 3, 2, 4), - nrow = 2, - ncol = 2 - ), - widths = c(5, 1, 5, 1), - heights = c(5, 1, 1, 1) - ) - opar <- par(mar = c(5, 5, 2.5, 2)) - on.exit(par(opar)) - plot( - MBordertable[, 1], - MBordertable[, 2], - xlab = "PC1", - ylab = "PC2", - pch = 20, - cex = 0, - col = "grey", - las = 1 - ) - for (k in 1:length(v)) { - points( - MBordertable[k, 1], - MBordertable[k, 2], - col = ColorRamp[v[k]], - pch = 20, - cex = 2 - ) - } - opar <- par(mar = c(3, 2.5, 2.5, 2)) - on.exit(par(opar)) - image( - 1, - ColorLevels, - matrix( - data = ColorLevels, - ncol = length(ColorLevels), - nrow = 1 - ), - col = ColorRamp, - xlab = "", - ylab = "", - las = 2, - xaxt = "n" - ) - layout(1) -} \ No newline at end of file diff --git a/R/PlotMBpca.R b/R/PlotMBpca.R new file mode 100644 index 0000000..17fb30d --- /dev/null +++ b/R/PlotMBpca.R @@ -0,0 +1,100 @@ +#' @title Plotting pseudo-time ordering or gene expression in Model-based clustering in PCA +#' @description The PCA representation can either be used to show pseudo-time ordering or the gene expression of a particular gene. +#' @param object \code{DISCBIO} class object. +#' @param type either `order` to plot pseudo-time ordering or `exp` to plot gene expression +#' @param g Individual gene name or vector with a group of gene names +#' corresponding to a subset of valid row names of the \code{ndata} slot of +#' the \code{DISCBIO} object. Ignored if `type="order"`. +#' @param n String of characters representing the title of the plot. Default is +#' NULL and the first element of \code{g} is chosen. Ignored if +#' `type="order"`. +#' @importFrom RColorBrewer brewer.pal +#' @importFrom grDevices colorRampPalette +#' @importFrom graphics layout par image +#' @return A plot of the PCA. +#' @export + +PlotMBpca <- function(object, type="order", g=NULL, n=NULL) { + # ========================================================================== + # Validation + # ========================================================================== + data <- object@MBclusters + if (type == "exp") { + if (is.null(g)) { + stop('g must be provided if type="exp"') + } + if (length(intersect(g, rownames(object@ndata))) < length(unique(g))) { + stop( + "second argument does not correspond to set of rownames slot", + "ndata of SCseq object" + ) + } + if (is.null(n)) { + n <- g[1] + } + l <- apply(object@ndata[g,] - .1, 2, sum) + .1 + x <- data$pcareduceres + } else if (type == "order") { + MBordertable <- cbind(data$pcareduceres, object@MBordering) + l <- MBordertable[, 3] + x <- MBordertable + } else { + stop("Invalid type. Valid alternatives as 'order' and 'exp'.") + } + # ========================================================================== + # Plotting + # ========================================================================== + mi <- min(l, na.rm = TRUE) + ma <- max(l, na.rm = TRUE) + ColorRamp <- colorRampPalette(rev(brewer.pal(n = 11, name = "RdYlBu")))(100) + ColorLevels <- seq(mi, ma, length = length(ColorRamp)) + v <- round((l - mi) / (ma - mi) * 99 + 1, 0) + layout( + matrix( + data = c(1, 3, 2, 4), + nrow = 2, + ncol = 2 + ), + widths = c(5, 1, 5, 1), + heights = c(5, 1, 1, 1) + ) + opar <- par(mar = c(5, 5, 2.5, 2)) + on.exit(par(opar)) + plot( + x[, 1], + x[, 2], + xlab = "PC1", + ylab = "PC2", + pch = 20, + cex = 0, + col = "grey", + las = 1, + main = n + ) + for (k in 1:length(v)) { + points( + x[k, 1], + x[k, 2], + col = ColorRamp[v[k]], + pch = 20, + cex = 2 + ) + } + opar <- par(mar = c(3, 2.5, 2.5, 2)) + on.exit(par(opar)) + image( + 1, + ColorLevels, + matrix( + data = ColorLevels, + ncol = length(ColorLevels), + nrow = 1 + ), + col = ColorRamp, + xlab = "", + ylab = "", + las = 2, + xaxt = "n" + ) + layout(1) +} \ No newline at end of file diff --git a/man/PlotMBexpPCA.Rd b/man/PlotMBexpPCA.Rd deleted file mode 100644 index f946b93..0000000 --- a/man/PlotMBexpPCA.Rd +++ /dev/null @@ -1,25 +0,0 @@ -% Generated by roxygen2: do not edit by hand -% Please edit documentation in R/PlotMBexpPCA.R -\name{PlotMBexpPCA} -\alias{PlotMBexpPCA} -\title{Plotting gene expression in Model-based clustering in PCA.} -\usage{ -PlotMBexpPCA(object, g, n = NULL) -} -\arguments{ -\item{object}{\code{DISCBIO} class object.} - -\item{g}{Individual gene name or vector with a group of gene names -corresponding to a subset of valid row names of the \code{ndata} slot of -the \code{DISCBIO} object.} - -\item{n}{String of characters representing the title of the plot. Default is -NULL and the first element of \code{g} is chosen.} -} -\value{ -A plot of the PCA. -} -\description{ -The PCA representation can also be used to show the gene - expression of a particular gene. -} diff --git a/man/PlotMBorderPCA.Rd b/man/PlotMBorderPCA.Rd deleted file mode 100644 index cdc4775..0000000 --- a/man/PlotMBorderPCA.Rd +++ /dev/null @@ -1,18 +0,0 @@ -% Generated by roxygen2: do not edit by hand -% Please edit documentation in R/PlotMBorderPCA.R -\name{PlotMBorderPCA} -\alias{PlotMBorderPCA} -\title{Plotting pseudo-time ordering in Model-based clustering in PCA.} -\usage{ -PlotMBorderPCA(object) -} -\arguments{ -\item{object}{\code{DISCBIO} class object.} -} -\value{ -A plot of the PCA. -} -\description{ -The PCA representation can also be used to show the pseudo-time - ordering. -} diff --git a/man/PlotMBpca.Rd b/man/PlotMBpca.Rd new file mode 100644 index 0000000..8057c50 --- /dev/null +++ b/man/PlotMBpca.Rd @@ -0,0 +1,27 @@ +% Generated by roxygen2: do not edit by hand +% Please edit documentation in R/PlotMBpca.R +\name{PlotMBpca} +\alias{PlotMBpca} +\title{Plotting pseudo-time ordering or gene expression in Model-based clustering in PCA} +\usage{ +PlotMBpca(object, type = "order", g = NULL, n = NULL) +} +\arguments{ +\item{object}{\code{DISCBIO} class object.} + +\item{type}{either `order` to plot pseudo-time ordering or `exp` to plot gene expression} + +\item{g}{Individual gene name or vector with a group of gene names +corresponding to a subset of valid row names of the \code{ndata} slot of +the \code{DISCBIO} object. Ignored if `type="order"`.} + +\item{n}{String of characters representing the title of the plot. Default is +NULL and the first element of \code{g} is chosen. Ignored if +`type="order"`.} +} +\value{ +A plot of the PCA. +} +\description{ +The PCA representation can either be used to show pseudo-time ordering or the gene expression of a particular gene. +} From 67aafb287c29dd8dc54691118884d4987760f6ca Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 3 Aug 2020 11:55:54 +0200 Subject: [PATCH 71/99] Merged plotOrderKMtsne and plotOrderMBtsne --- DESCRIPTION | 3 +- NAMESPACE | 3 +- R/DIscBIO-generic-plotOrderMBtsne.R | 79 ------------------- ...tsne.R => DIscBIO-generic-plotOrderTsne.R} | 44 +++++++---- man/plotOrderKMtsne.Rd | 22 ------ man/plotOrderMBtsne.Rd | 22 ------ man/plotOrderTsne.Rd | 21 +++++ 7 files changed, 51 insertions(+), 143 deletions(-) delete mode 100644 R/DIscBIO-generic-plotOrderMBtsne.R rename R/{DIscBIO-generic-plotOrderKMtsne.R => DIscBIO-generic-plotOrderTsne.R} (65%) delete mode 100644 man/plotOrderKMtsne.Rd delete mode 100644 man/plotOrderMBtsne.Rd create mode 100644 man/plotOrderTsne.Rd diff --git a/DESCRIPTION b/DESCRIPTION index ed73306..ba5795e 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -79,8 +79,7 @@ Collate: 'DIscBIO-generic-plotGap.R' 'DIscBIO-generic-plotKmeansLabelstSNE.R' 'DIscBIO-generic-plotMBLabelstSNE.R' - 'DIscBIO-generic-plotOrderKMtsne.R' - 'DIscBIO-generic-plotOrderMBtsne.R' + 'DIscBIO-generic-plotOrderTsne.R' 'DIscBIO-generic-plotSilhouette.R' 'DIscBIO-generic-plotSymbolstSNE.R' 'DIscBIO-generic-plotexptsneMB.R' diff --git a/NAMESPACE b/NAMESPACE index f319841..5b5c63c 100644 --- a/NAMESPACE +++ b/NAMESPACE @@ -41,8 +41,7 @@ exportMethods(comptsneMB) exportMethods(plotExptSNE) exportMethods(plotKmeansLabelstSNE) exportMethods(plotMBLabelstSNE) -exportMethods(plotOrderKMtsne) -exportMethods(plotOrderMBtsne) +exportMethods(plotOrderTsne) exportMethods(plotSilhouette) exportMethods(plotSymbolstSNE) exportMethods(plotexptsneMB) diff --git a/R/DIscBIO-generic-plotOrderMBtsne.R b/R/DIscBIO-generic-plotOrderMBtsne.R deleted file mode 100644 index 22920ca..0000000 --- a/R/DIscBIO-generic-plotOrderMBtsne.R +++ /dev/null @@ -1,79 +0,0 @@ -#' @title Plotting the pseudo-time ordering based on Model-based clusters in the -#' t-SNE map -#' @description The tSNE representation can also be used to show the pseudo-time -#' ordering. -#' @param object \code{DISCBIO} class object. -#' @return A plot of the pseudo-time ordering. -setGeneric("plotOrderMBtsne", function(object) - standardGeneric("plotOrderMBtsne")) - -#' @export -#' @rdname plotOrderMBtsne -setMethod( - "plotOrderMBtsne", - signature = "DISCBIO", - definition = function(object) { - if (length(object@MBtsne) == 0) - stop("run comptsneMB before plotOrderMBtsne") - total <- rbind(object@ndata, object@MBordering) - rownames(total)[nrow(total)] <- - "Pseudo-time ordering of MBclustering" - g <- rownames(total)[nrow(total)] - n <- g[1] - l <- apply(total[g, ] - .1, 2, sum) + .1 - - mi <- min(l, na.rm = TRUE) - ma <- max(l, na.rm = TRUE) - ColorRamp <- - colorRampPalette(rev(brewer.pal(n = 7, name = "RdYlBu")))(100) - ColorLevels <- seq(mi, ma, length = length(ColorRamp)) - v <- round((l - mi) / (ma - mi) * 99 + 1, 0) - layout( - matrix( - data = c(1, 3, 2, 4), - nrow = 2, - ncol = 2 - ), - widths = c(5, 1, 5, 1), - heights = c(5, 1, 1, 1) - ) - opar <- par(mar = c(3, 5, 2.5, 2)) - on.exit(par(opar)) - plot( - object@MBtsne, - xlab = "Dim 1", - ylab = "Dim 2", - main = n, - pch = 20, - cex = 0, - col = "grey", - las = 1 - ) - for (k in 1:length(v)) { - points( - object@MBtsne[k, 1], - object@MBtsne[k, 2], - col = ColorRamp[v[k]], - pch = 20, - cex = 1.5 - ) - } - opar <- par(mar = c(3, 2.5, 2.5, 2)) - on.exit(par(opar)) - image( - 1, - ColorLevels, - matrix( - data = ColorLevels, - ncol = length(ColorLevels), - nrow = 1 - ), - col = ColorRamp, - xlab = "", - ylab = "", - las = 1, - xaxt = "n" - ) - layout(1) - } -) \ No newline at end of file diff --git a/R/DIscBIO-generic-plotOrderKMtsne.R b/R/DIscBIO-generic-plotOrderTsne.R similarity index 65% rename from R/DIscBIO-generic-plotOrderKMtsne.R rename to R/DIscBIO-generic-plotOrderTsne.R index d1d5f72..adeb4f8 100644 --- a/R/DIscBIO-generic-plotOrderKMtsne.R +++ b/R/DIscBIO-generic-plotOrderTsne.R @@ -1,31 +1,43 @@ -#' @title Plotting the pseudo-time ordering based on K-means clusters in the -#' t-SNE map +#' @title Plotting the pseudo-time ordering in the t-SNE map #' @description The tSNE representation can also be used to show the pseudo-time #' ordering. #' @param object \code{DISCBIO} class object. #' @return A plot of the pseudo-time ordering. -setGeneric("plotOrderKMtsne", function(object) - standardGeneric("plotOrderKMtsne")) +setGeneric("plotOrderTsne", function(object) + standardGeneric("plotOrderTsne")) #' @export -#' @rdname plotOrderKMtsne +#' @rdname plotOrderTsne setMethod( - "plotOrderKMtsne", + "plotOrderTsne", signature = "DISCBIO", definition = function(object) { - if (length(object@tsne) == 0) - stop("run comptsne before plotOrderKMtsne") - total <- rbind(object@ndata, object@kordering) - rownames(total)[nrow(total)] <- - "Pseudo-time ordering of K-means clustering" + ran_k <- length(object@tsne) > 0 + ran_m <- length(object@MBtsne) > 0 + if (ran_k) { + total <- rbind(object@ndata, object@kordering) + clustering_method <- "k-means" + x <- object@tsne + } else if (ran_m) { + total <- rbind(object@ndata, object@MBordering) + clustering_method <- "model-based" + x <- object@MBtsne + } else { + stop("run comptsne or comptsneMB before plotOrderTsne") + } + + rownames(total)[nrow(total)] <- paste( + "Pseudo-time ordering of", clustering_method, "clustering" + ) g <- rownames(total)[nrow(total)] n <- g[1] l <- apply(total[g, ] - .1, 2, sum) + .1 mi <- min(l, na.rm = TRUE) ma <- max(l, na.rm = TRUE) - ColorRamp <- - colorRampPalette(rev(brewer.pal(n = 7, name = "RdYlBu")))(100) + ColorRamp <- colorRampPalette( + rev(brewer.pal(n = 7, name = "RdYlBu")) + )(100) ColorLevels <- seq(mi, ma, length = length(ColorRamp)) v <- round((l - mi) / (ma - mi) * 99 + 1, 0) layout( @@ -40,7 +52,7 @@ setMethod( opar <- par(mar = c(3, 5, 2.5, 2)) on.exit(par(opar)) plot( - object@tsne, + x, xlab = "Dim 1", ylab = "Dim 2", main = n, @@ -51,8 +63,8 @@ setMethod( ) for (k in 1:length(v)) { points( - object@tsne[k, 1], - object@tsne[k, 2], + x[k, 1], + x[k, 2], col = ColorRamp[v[k]], pch = 20, cex = 1.5 diff --git a/man/plotOrderKMtsne.Rd b/man/plotOrderKMtsne.Rd deleted file mode 100644 index 7d220eb..0000000 --- a/man/plotOrderKMtsne.Rd +++ /dev/null @@ -1,22 +0,0 @@ -% Generated by roxygen2: do not edit by hand -% Please edit documentation in R/DIscBIO-generic-plotOrderKMtsne.R -\name{plotOrderKMtsne} -\alias{plotOrderKMtsne} -\alias{plotOrderKMtsne,DISCBIO-method} -\title{Plotting the pseudo-time ordering based on K-means clusters in the - t-SNE map} -\usage{ -plotOrderKMtsne(object) - -\S4method{plotOrderKMtsne}{DISCBIO}(object) -} -\arguments{ -\item{object}{\code{DISCBIO} class object.} -} -\value{ -A plot of the pseudo-time ordering. -} -\description{ -The tSNE representation can also be used to show the pseudo-time - ordering. -} diff --git a/man/plotOrderMBtsne.Rd b/man/plotOrderMBtsne.Rd deleted file mode 100644 index fdd099e..0000000 --- a/man/plotOrderMBtsne.Rd +++ /dev/null @@ -1,22 +0,0 @@ -% Generated by roxygen2: do not edit by hand -% Please edit documentation in R/DIscBIO-generic-plotOrderMBtsne.R -\name{plotOrderMBtsne} -\alias{plotOrderMBtsne} -\alias{plotOrderMBtsne,DISCBIO-method} -\title{Plotting the pseudo-time ordering based on Model-based clusters in the - t-SNE map} -\usage{ -plotOrderMBtsne(object) - -\S4method{plotOrderMBtsne}{DISCBIO}(object) -} -\arguments{ -\item{object}{\code{DISCBIO} class object.} -} -\value{ -A plot of the pseudo-time ordering. -} -\description{ -The tSNE representation can also be used to show the pseudo-time - ordering. -} diff --git a/man/plotOrderTsne.Rd b/man/plotOrderTsne.Rd new file mode 100644 index 0000000..f53b524 --- /dev/null +++ b/man/plotOrderTsne.Rd @@ -0,0 +1,21 @@ +% Generated by roxygen2: do not edit by hand +% Please edit documentation in R/DIscBIO-generic-plotOrderTsne.R +\name{plotOrderTsne} +\alias{plotOrderTsne} +\alias{plotOrderTsne,DISCBIO-method} +\title{Plotting the pseudo-time ordering in the t-SNE map} +\usage{ +plotOrderTsne(object) + +\S4method{plotOrderTsne}{DISCBIO}(object) +} +\arguments{ +\item{object}{\code{DISCBIO} class object.} +} +\value{ +A plot of the pseudo-time ordering. +} +\description{ +The tSNE representation can also be used to show the pseudo-time + ordering. +} From ca06837029725fc29e045a6566ae1ed8221bba2b Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 3 Aug 2020 13:29:34 +0200 Subject: [PATCH 72/99] Merged comptSNE and comptsneMB --- DESCRIPTION | 1 - NAMESPACE | 1 - R/DIscBIO-generic-comptSNE.R | 63 +++++++++++++++++++--------- R/DIscBIO-generic-comptsneMB.R | 43 ------------------- R/DIscBIO-generic-plotMBLabelstSNE.R | 2 +- R/DIscBIO-generic-plotOrderTsne.R | 2 +- R/DIscBIO-generic-plotexptsneMB.R | 2 +- R/DIscBIO-generic-plottsneMB.R | 2 +- R/prepExampleDataset.R | 2 +- man/comptSNE.Rd | 2 +- man/comptsneMB.Rd | 45 -------------------- 11 files changed, 49 insertions(+), 116 deletions(-) delete mode 100644 R/DIscBIO-generic-comptsneMB.R delete mode 100644 man/comptsneMB.Rd diff --git a/DESCRIPTION b/DESCRIPTION index ba5795e..c4a8cd5 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -74,7 +74,6 @@ Collate: 'DIscBIO-generic-PCAplotSymbols.R' 'DIscBIO-generic-PlotmclustMB.R' 'DIscBIO-generic-comptSNE.R' - 'DIscBIO-generic-comptsneMB.R' 'DIscBIO-generic-plotExptSNE.R' 'DIscBIO-generic-plotGap.R' 'DIscBIO-generic-plotKmeansLabelstSNE.R' diff --git a/NAMESPACE b/NAMESPACE index 5b5c63c..2c32c79 100644 --- a/NAMESPACE +++ b/NAMESPACE @@ -37,7 +37,6 @@ exportMethods(Normalizedata) exportMethods(PCAplotSymbols) exportMethods(PlotmclustMB) exportMethods(comptSNE) -exportMethods(comptsneMB) exportMethods(plotExptSNE) exportMethods(plotKmeansLabelstSNE) exportMethods(plotMBLabelstSNE) diff --git a/R/DIscBIO-generic-comptSNE.R b/R/DIscBIO-generic-comptSNE.R index 71754d4..acb7a34 100644 --- a/R/DIscBIO-generic-comptSNE.R +++ b/R/DIscBIO-generic-comptSNE.R @@ -1,4 +1,4 @@ -#' @title Computing tSNE for K-means clustering +#' @title Computing tSNE #' @description This function is used to compute the t-Distributed Stochastic #' Neighbor Embedding (t-SNE). #' @param object \code{DISCBIO} class object. @@ -14,30 +14,53 @@ #' setGeneric( name = "comptSNE", - def = function(object, rseed = NULL, max_iter = 5000, epoch = 500, - quiet = FALSE, ...) { + def = function( + object, rseed=NULL, max_iter=5000, epoch=500, quiet=FALSE, ... + ) + { standardGeneric("comptSNE") } ) #' @rdname comptSNE #' @export - setMethod( - "comptSNE", +setMethod( + f = "comptSNE", signature = "DISCBIO", - definition = function(object, rseed, max_iter, epoch, quiet, ...) { - if (length(object@kmeans$kpart) == 0) - stop("run Clustexp before comptSNE") - set.seed(rseed) - di <- dist.gen(as.matrix(object@distances)) - if (quiet) { - ts <- suppressMessages( - tsne(di, max_iter = max_iter, epoch = epoch, ...) - ) - } else { - ts <- tsne(di, max_iter = max_iter, epoch = epoch, ...) + definition = function(object, rseed, max_iter, epoch, quiet, ...) + { + # ====================================================================== + # Validating + # ====================================================================== + ran_k <- length(object@kmeans$kpart) > 0 + ran_m <- length(object@MBclusters) > 0 + if (ran_k) { + di <- dist.gen(as.matrix(object@distances)) + } else if (ran_m) { + di <- dist.gen(as.matrix(t(object@fdata))) + } else { + stop("run clustexp before comptSNE") + } + # ====================================================================== + # Computing + # ====================================================================== + set.seed(rseed) + if (quiet) { + ts <- suppressMessages( + tsne(di, max_iter = max_iter, epoch = epoch, ...) + ) + } else { + message("This function may take time") + ts <- tsne(di, max_iter = max_iter, epoch = epoch, ...) + } + # ====================================================================== + # Filling output + # ====================================================================== + if (ran_k) { + object@tsne <- as.data.frame(ts) + } else if (ran_m) { + object@MBtsne <- as.data.frame(ts) + } + return(object) } - object@tsne <- as.data.frame(ts) - return(object) - } - ) \ No newline at end of file +) \ No newline at end of file diff --git a/R/DIscBIO-generic-comptsneMB.R b/R/DIscBIO-generic-comptsneMB.R deleted file mode 100644 index 7c0ae18..0000000 --- a/R/DIscBIO-generic-comptsneMB.R +++ /dev/null @@ -1,43 +0,0 @@ -#' @title Computing tSNE for Model-based clustering -#' @description This function is used to compute the t-Distributed Stochastic -#' Neighbor Embedding (t-SNE). -#' @param object \code{DISCBIO} class object. -#' @param max_iter maximum number of iterations to perform. -#' @param epoch The number of iterations in between update messages. -#' @param quiet if `TRUE`, suppresses intermediate output. -#' @param rseed Random number to be set for reproducible results. -#' @param ... other parameters to be passed to `tsne::tsne`. -#' @importFrom tsne tsne -#' @importFrom stats as.dist cor -#' @return The DISCBIO-class object input with the MBtsne slot filled. -#' -setGeneric( - name = "comptsneMB", - def = function(object, rseed = NULL, max_iter = 5000, epoch = 500, - quiet = FALSE, ...) { - standardGeneric("comptsneMB") - } -) - -#' @rdname comptsneMB -#' @export -setMethod( - f = "comptsneMB", - signature = "DISCBIO", - definition = function(object, rseed, max_iter, epoch, quiet, ...) { - if (length(object@MBclusters) == 0) - stop("run clustexp before comptsneMB") - set.seed(rseed) - di <- dist.gen(as.matrix(t(object@fdata))) - if (quiet) { - ts <- suppressMessages( - tsne(di, max_iter = max_iter, epoch = epoch, ...) - ) - } else { - message("This function may take time") - ts <- tsne(di, max_iter = max_iter, epoch = epoch, ...) - } - object@MBtsne <- as.data.frame(ts) - return(object) - } -) \ No newline at end of file diff --git a/R/DIscBIO-generic-plotMBLabelstSNE.R b/R/DIscBIO-generic-plotMBLabelstSNE.R index 71d52c3..d3d4f26 100644 --- a/R/DIscBIO-generic-plotMBLabelstSNE.R +++ b/R/DIscBIO-generic-plotMBLabelstSNE.R @@ -14,7 +14,7 @@ setMethod( signature = "DISCBIO", definition = function(object) { if (length(object@MBtsne) == 0) - stop("run comptsneMB before plotMBLabelstSNE") + stop("run comptsne before plotMBLabelstSNE") Clusters <- object@MBclusters$clusterid ClustersFactor <- as.factor(Clusters) ClustersFactor <- gsub("1", "black", ClustersFactor) diff --git a/R/DIscBIO-generic-plotOrderTsne.R b/R/DIscBIO-generic-plotOrderTsne.R index adeb4f8..1b6c96e 100644 --- a/R/DIscBIO-generic-plotOrderTsne.R +++ b/R/DIscBIO-generic-plotOrderTsne.R @@ -23,7 +23,7 @@ setMethod( clustering_method <- "model-based" x <- object@MBtsne } else { - stop("run comptsne or comptsneMB before plotOrderTsne") + stop("run comptsne before plotOrderTsne") } rownames(total)[nrow(total)] <- paste( diff --git a/R/DIscBIO-generic-plotexptsneMB.R b/R/DIscBIO-generic-plotexptsneMB.R index 6e239e7..dcb9b38 100644 --- a/R/DIscBIO-generic-plotexptsneMB.R +++ b/R/DIscBIO-generic-plotexptsneMB.R @@ -20,7 +20,7 @@ setMethod( signature = "DISCBIO", definition = function(object, g, n = NULL) { if (length(object@MBtsne) == 0) - stop("run comptsneMB before plotexptsneMB") + stop("run comptsne before plotexptsneMB") if (length(intersect(g, rownames(object@ndata))) < length(unique(g))) stop( "second argument does not correspond to set of rownames", diff --git a/R/DIscBIO-generic-plottsneMB.R b/R/DIscBIO-generic-plottsneMB.R index 64452ed..00140be 100644 --- a/R/DIscBIO-generic-plottsneMB.R +++ b/R/DIscBIO-generic-plottsneMB.R @@ -18,7 +18,7 @@ setMethod( signature = "DISCBIO", definition = function(object, K) { if (length(object@MBtsne) == 0) - stop("run comptsneMB before plottsneMB") + stop("run comptsne before plottsneMB") col = c("black", "blue", "green", "red", "yellow", "gray") part <- object@MBclusters$clusterid plot( diff --git a/R/prepExampleDataset.R b/R/prepExampleDataset.R index e834c58..336a1ff 100644 --- a/R/prepExampleDataset.R +++ b/R/prepExampleDataset.R @@ -29,7 +29,7 @@ prepExampleDataset <- function(dataset, save=TRUE) { valuesG1msReduced_treated_K <- sc_k message("Model-based clustering") sc_mb <- Exprmclust(sc, quiet=TRUE) - sc_mb <- comptsneMB(sc_mb, rseed=15555, quiet=TRUE) + sc_mb <- comptSNE(sc_mb, rseed=15555, quiet=TRUE) valuesG1msReduced_treated_MB <- sc_mb # ========================================================================== # Output diff --git a/man/comptSNE.Rd b/man/comptSNE.Rd index 3d80f40..e79add1 100644 --- a/man/comptSNE.Rd +++ b/man/comptSNE.Rd @@ -3,7 +3,7 @@ \name{comptSNE} \alias{comptSNE} \alias{comptSNE,DISCBIO-method} -\title{Computing tSNE for K-means clustering} +\title{Computing tSNE} \usage{ comptSNE( object, diff --git a/man/comptsneMB.Rd b/man/comptsneMB.Rd deleted file mode 100644 index ec623b3..0000000 --- a/man/comptsneMB.Rd +++ /dev/null @@ -1,45 +0,0 @@ -% Generated by roxygen2: do not edit by hand -% Please edit documentation in R/DIscBIO-generic-comptsneMB.R -\name{comptsneMB} -\alias{comptsneMB} -\alias{comptsneMB,DISCBIO-method} -\title{Computing tSNE for Model-based clustering} -\usage{ -comptsneMB( - object, - rseed = NULL, - max_iter = 5000, - epoch = 500, - quiet = FALSE, - ... -) - -\S4method{comptsneMB}{DISCBIO}( - object, - rseed = NULL, - max_iter = 5000, - epoch = 500, - quiet = FALSE, - ... -) -} -\arguments{ -\item{object}{\code{DISCBIO} class object.} - -\item{rseed}{Random number to be set for reproducible results.} - -\item{max_iter}{maximum number of iterations to perform.} - -\item{epoch}{The number of iterations in between update messages.} - -\item{quiet}{if `TRUE`, suppresses intermediate output.} - -\item{...}{other parameters to be passed to `tsne::tsne`.} -} -\value{ -The DISCBIO-class object input with the MBtsne slot filled. -} -\description{ -This function is used to compute the t-Distributed Stochastic - Neighbor Embedding (t-SNE). -} From 711de04a03d0cbdd0b4569e6efed0463f54540c5 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 3 Aug 2020 13:35:29 +0200 Subject: [PATCH 73/99] Merged plotKmeansLabelstSNE and plotMBLabelstSNE --- DESCRIPTION | 3 +- NAMESPACE | 3 +- R/DIscBIO-generic-plotKmeansLabelstSNE.R | 44 ----------------- R/DIscBIO-generic-plotLabelstSNE.R | 62 ++++++++++++++++++++++++ R/DIscBIO-generic-plotMBLabelstSNE.R | 44 ----------------- man/plotKmeansLabelstSNE.Rd | 20 -------- man/plotLabelstSNE.Rd | 20 ++++++++ man/plotMBLabelstSNE.Rd | 20 -------- 8 files changed, 84 insertions(+), 132 deletions(-) delete mode 100644 R/DIscBIO-generic-plotKmeansLabelstSNE.R create mode 100644 R/DIscBIO-generic-plotLabelstSNE.R delete mode 100644 R/DIscBIO-generic-plotMBLabelstSNE.R delete mode 100644 man/plotKmeansLabelstSNE.Rd create mode 100644 man/plotLabelstSNE.Rd delete mode 100644 man/plotMBLabelstSNE.Rd diff --git a/DESCRIPTION b/DESCRIPTION index c4a8cd5..9e64b63 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -76,8 +76,7 @@ Collate: 'DIscBIO-generic-comptSNE.R' 'DIscBIO-generic-plotExptSNE.R' 'DIscBIO-generic-plotGap.R' - 'DIscBIO-generic-plotKmeansLabelstSNE.R' - 'DIscBIO-generic-plotMBLabelstSNE.R' + 'DIscBIO-generic-plotLabelstSNE.R' 'DIscBIO-generic-plotOrderTsne.R' 'DIscBIO-generic-plotSilhouette.R' 'DIscBIO-generic-plotSymbolstSNE.R' diff --git a/NAMESPACE b/NAMESPACE index 2c32c79..2acaf4a 100644 --- a/NAMESPACE +++ b/NAMESPACE @@ -38,8 +38,7 @@ exportMethods(PCAplotSymbols) exportMethods(PlotmclustMB) exportMethods(comptSNE) exportMethods(plotExptSNE) -exportMethods(plotKmeansLabelstSNE) -exportMethods(plotMBLabelstSNE) +exportMethods(plotLabelstSNE) exportMethods(plotOrderTsne) exportMethods(plotSilhouette) exportMethods(plotSymbolstSNE) diff --git a/R/DIscBIO-generic-plotKmeansLabelstSNE.R b/R/DIscBIO-generic-plotKmeansLabelstSNE.R deleted file mode 100644 index a033449..0000000 --- a/R/DIscBIO-generic-plotKmeansLabelstSNE.R +++ /dev/null @@ -1,44 +0,0 @@ -#' @title tSNE map for K-means clustering with labels -#' @description Visualizing the K-means clusters using tSNE maps -#' @param object \code{DISCBIO} class object. -#' @rdname plotKmeansLabelstSNE -#' @importFrom graphics text -#' @return Plot containing the ID of the cells in each cluster -setGeneric("plotKmeansLabelstSNE", function(object) - standardGeneric("plotKmeansLabelstSNE")) - -#' @rdname plotKmeansLabelstSNE -#' @export -setMethod( - "plotKmeansLabelstSNE", - signature = "DISCBIO", - definition = function(object) { - if (length(object@tsne) == 0) - stop("run comptsne before plotKmeansLabelstSNE") - Clusters <- object@kmeans$kpart - ClustersFactor <- as.factor(Clusters) - ClustersFactor <- gsub("1", "black", ClustersFactor) - ClustersFactor <- gsub("2", "blue", ClustersFactor) - ClustersFactor <- gsub("3", "green", ClustersFactor) - ClustersFactor <- gsub("4", "red", ClustersFactor) - ClustersFactor <- gsub("5", "yellow", ClustersFactor) - ClustersFactor <- gsub("6", "gray", ClustersFactor) - COL <- ClustersFactor - labels = names(object@ndata) - plot( - object@tsne, - xlab = "Dim 1", - ylab = "Dim 2", - pch = 20, - cex = .5, - col = "lightgrey" - ) - text( - object@tsne[, 1], - object@tsne[, 2], - labels, - cex = .7, - col = COL - ) - } -) \ No newline at end of file diff --git a/R/DIscBIO-generic-plotLabelstSNE.R b/R/DIscBIO-generic-plotLabelstSNE.R new file mode 100644 index 0000000..41c9f8e --- /dev/null +++ b/R/DIscBIO-generic-plotLabelstSNE.R @@ -0,0 +1,62 @@ +#' @title tSNE map with labels +#' @description Visualizing k-means or model-based clusters using tSNE maps +#' @param object \code{DISCBIO} class object. +#' @rdname plotLabelstSNE +#' @importFrom graphics text +#' @return Plot containing the ID of the cells in each cluster +setGeneric("plotLabelstSNE", function(object) + { + standardGeneric("plotLabelstSNE") + } +) + +#' @rdname plotLabelstSNE +#' @export +setMethod( + "plotLabelstSNE", + signature = "DISCBIO", + definition = function(object) + { + # ====================================================================== + # Validating + # ====================================================================== + ran_k <- length(object@tsne) > 0 + ran_m <- length(object@MBtsne) > 0 + if (ran_k) { + Clusters <- object@kmeans$kpart + x <- object@tsne + } else if (ran_m) { + Clusters <- object@MBclusters$clusterid + x <- object@MBtsne + } else { + stop("run comptsne before plotLabelstSNE") + } + # ====================================================================== + # Plotting + # ====================================================================== + ClustersFactor <- as.factor(Clusters) + ClustersFactor <- gsub("1", "black", ClustersFactor) + ClustersFactor <- gsub("2", "blue", ClustersFactor) + ClustersFactor <- gsub("3", "green", ClustersFactor) + ClustersFactor <- gsub("4", "red", ClustersFactor) + ClustersFactor <- gsub("5", "yellow", ClustersFactor) + ClustersFactor <- gsub("6", "gray", ClustersFactor) + COL <- ClustersFactor + labels = names(object@ndata) + plot( + x, + xlab = "Dim 1", + ylab = "Dim 2", + pch = 20, + cex = .5, + col = "lightgrey" + ) + text( + x[, 1], + x[, 2], + labels, + cex = .7, + col = COL + ) + } +) \ No newline at end of file diff --git a/R/DIscBIO-generic-plotMBLabelstSNE.R b/R/DIscBIO-generic-plotMBLabelstSNE.R deleted file mode 100644 index d3d4f26..0000000 --- a/R/DIscBIO-generic-plotMBLabelstSNE.R +++ /dev/null @@ -1,44 +0,0 @@ -#' @title tSNE map for Model-based clustering with labels -#' @description Visualizing the Model-based clusters using tSNE maps -#' @param object \code{DISCBIO} class object. -#' @importFrom graphics text -#' @return A plot of the `object@MBtsne` values -setGeneric("plotMBLabelstSNE", function(object) - standardGeneric("plotMBLabelstSNE") -) - -#' @rdname plotMBLabelstSNE -#' @export -setMethod( - "plotMBLabelstSNE", - signature = "DISCBIO", - definition = function(object) { - if (length(object@MBtsne) == 0) - stop("run comptsne before plotMBLabelstSNE") - Clusters <- object@MBclusters$clusterid - ClustersFactor <- as.factor(Clusters) - ClustersFactor <- gsub("1", "black", ClustersFactor) - ClustersFactor <- gsub("2", "blue", ClustersFactor) - ClustersFactor <- gsub("3", "green", ClustersFactor) - ClustersFactor <- gsub("4", "red", ClustersFactor) - ClustersFactor <- gsub("5", "yellow", ClustersFactor) - ClustersFactor <- gsub("6", "gray", ClustersFactor) - COL <- ClustersFactor - labels = names(object@ndata) - plot( - object@MBtsne, - xlab = "Dim 1", - ylab = "Dim 2", - pch = 20, - cex = .5, - col = "lightgrey" - ) - text( - object@MBtsne[, 1], - object@MBtsne[, 2], - labels, - cex = .7, - col = COL - ) - } -) \ No newline at end of file diff --git a/man/plotKmeansLabelstSNE.Rd b/man/plotKmeansLabelstSNE.Rd deleted file mode 100644 index c6185aa..0000000 --- a/man/plotKmeansLabelstSNE.Rd +++ /dev/null @@ -1,20 +0,0 @@ -% Generated by roxygen2: do not edit by hand -% Please edit documentation in R/DIscBIO-generic-plotKmeansLabelstSNE.R -\name{plotKmeansLabelstSNE} -\alias{plotKmeansLabelstSNE} -\alias{plotKmeansLabelstSNE,DISCBIO-method} -\title{tSNE map for K-means clustering with labels} -\usage{ -plotKmeansLabelstSNE(object) - -\S4method{plotKmeansLabelstSNE}{DISCBIO}(object) -} -\arguments{ -\item{object}{\code{DISCBIO} class object.} -} -\value{ -Plot containing the ID of the cells in each cluster -} -\description{ -Visualizing the K-means clusters using tSNE maps -} diff --git a/man/plotLabelstSNE.Rd b/man/plotLabelstSNE.Rd new file mode 100644 index 0000000..9cdb721 --- /dev/null +++ b/man/plotLabelstSNE.Rd @@ -0,0 +1,20 @@ +% Generated by roxygen2: do not edit by hand +% Please edit documentation in R/DIscBIO-generic-plotLabelstSNE.R +\name{plotLabelstSNE} +\alias{plotLabelstSNE} +\alias{plotLabelstSNE,DISCBIO-method} +\title{tSNE map with labels} +\usage{ +plotLabelstSNE(object) + +\S4method{plotLabelstSNE}{DISCBIO}(object) +} +\arguments{ +\item{object}{\code{DISCBIO} class object.} +} +\value{ +Plot containing the ID of the cells in each cluster +} +\description{ +Visualizing k-means or model-based clusters using tSNE maps +} diff --git a/man/plotMBLabelstSNE.Rd b/man/plotMBLabelstSNE.Rd deleted file mode 100644 index 7b67822..0000000 --- a/man/plotMBLabelstSNE.Rd +++ /dev/null @@ -1,20 +0,0 @@ -% Generated by roxygen2: do not edit by hand -% Please edit documentation in R/DIscBIO-generic-plotMBLabelstSNE.R -\name{plotMBLabelstSNE} -\alias{plotMBLabelstSNE} -\alias{plotMBLabelstSNE,DISCBIO-method} -\title{tSNE map for Model-based clustering with labels} -\usage{ -plotMBLabelstSNE(object) - -\S4method{plotMBLabelstSNE}{DISCBIO}(object) -} -\arguments{ -\item{object}{\code{DISCBIO} class object.} -} -\value{ -A plot of the `object@MBtsne` values -} -\description{ -Visualizing the Model-based clusters using tSNE maps -} From 7581a363723e8c93112d31ea66d02f21c043c09c Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 3 Aug 2020 14:43:42 +0200 Subject: [PATCH 74/99] Removed samr dependency --- DESCRIPTION | 2 +- NAMESPACE | 5 +- R/DIscBIO-generic-DEGanalysis.R | 2 - R/DIscBIO-generic-DEGanalysis2clust.R | 2 - R/VolcanoPlot.R | 2 - R/samr-adapted.R | 739 ++++++++++++++++++++++++++ 6 files changed, 741 insertions(+), 11 deletions(-) diff --git a/DESCRIPTION b/DESCRIPTION index 9e64b63..a7162d4 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -37,7 +37,7 @@ Description: An open, multi-algorithmic pipeline for easy, fast and efficient . License: MIT + file LICENSE Encoding: UTF-8 -Imports: methods, TSCAN, boot, httr, mclust, statmod, samr, igraph, +Imports: methods, TSCAN, boot, httr, mclust, statmod, igraph, RWeka, philentropy, NetIndices, png, grDevices, readr, RColorBrewer, ggplot2, rpart, fpc, cluster, rpart.plot, tsne, AnnotationDbi, org.Hs.eg.db, graphics, stats, utils, impute diff --git a/NAMESPACE b/NAMESPACE index 2acaf4a..2ce6381 100644 --- a/NAMESPACE +++ b/NAMESPACE @@ -123,10 +123,6 @@ importFrom(png,readPNG) importFrom(readr,read_tsv) importFrom(rpart,rpart) importFrom(rpart.plot,rpart.plot) -importFrom(samr,samr) -importFrom(samr,samr.compute.delta.table) -importFrom(samr,samr.compute.siggenes.table) -importFrom(samr,samr.plot) importFrom(statmod,glmgam.fit) importFrom(stats,aggregate) importFrom(stats,as.dist) @@ -147,6 +143,7 @@ importFrom(stats,predict) importFrom(stats,quantile) importFrom(stats,rpois) importFrom(stats,runif) +importFrom(stats,smooth.spline) importFrom(stats,var) importFrom(tsne,tsne) importFrom(utils,capture.output) diff --git a/R/DIscBIO-generic-DEGanalysis.R b/R/DIscBIO-generic-DEGanalysis.R index ae2e637..c5f366f 100644 --- a/R/DIscBIO-generic-DEGanalysis.R +++ b/R/DIscBIO-generic-DEGanalysis.R @@ -15,8 +15,6 @@ #' @param plot if `TRUE`, plots are generated #' @param filename_deg Name of the exported DEG table #' @param filename_sigdeg Name of the exported sigDEG table -#' @importFrom samr samr samr.compute.delta.table samr.plot -#' samr.compute.siggenes.table #' @importFrom graphics title #' @importFrom utils write.csv capture.output #' @param ... additional parameters to be passed to samr() diff --git a/R/DIscBIO-generic-DEGanalysis2clust.R b/R/DIscBIO-generic-DEGanalysis2clust.R index b3e813d..a98a084 100644 --- a/R/DIscBIO-generic-DEGanalysis2clust.R +++ b/R/DIscBIO-generic-DEGanalysis2clust.R @@ -20,8 +20,6 @@ #' @param filename_deg Name of the exported DEG table #' @param filename_sigdeg Name of the exported sigDEG table #' @param ... additional parameters to be passed to samr() -#' @importFrom samr samr samr.compute.delta.table samr.plot -#' samr.compute.siggenes.table #' @importFrom graphics title #' @importFrom utils write.csv capture.output #' @importFrom AnnotationDbi keys diff --git a/R/VolcanoPlot.R b/R/VolcanoPlot.R index 5d19f89..c1c8398 100644 --- a/R/VolcanoPlot.R +++ b/R/VolcanoPlot.R @@ -9,8 +9,6 @@ #' @param fc A numeric value of the fold change. Default is 0.5. #' @param FS A numeric value of the font size. Default is 0.4. #' @param name A string vector showing the name to be used on the plot title -#' @importFrom samr samr samr.compute.delta.table samr.plot -#' samr.compute.siggenes.table #' @importFrom graphics title #' @importFrom utils write.csv #' @return A volcano plot diff --git a/R/samr-adapted.R b/R/samr-adapted.R index a4aca5a..a03c1aa 100644 --- a/R/samr-adapted.R +++ b/R/samr-adapted.R @@ -1378,3 +1378,742 @@ multiclass.seq.func <- function(xresamp, y) return(list(tt = tt, numer = tt, sd = rep(1, length(tt)), stand.contrasts = stand.contrasts)) } + +# ============================================================================== +# samr.compute.delta.table +# ============================================================================== +## Jun added starts +samr.compute.delta.table <- function(samr.obj, min.foldchange = 0, + dels = NULL, nvals = 50) { + res <- NULL + if (samr.obj$assay.type == "array") { + res <- samr.compute.delta.table.array(samr.obj, min.foldchange, + dels, nvals) + } + else if (samr.obj$assay.type == "seq") { + res <- samr.compute.delta.table.seq(samr.obj, min.foldchange, + dels) + } + return(res) +} +## Jun added ends + +## Jun added the first row below, and commented the row +# after it +samr.compute.delta.table.array <- function(samr.obj, + min.foldchange = 0, dels = NULL, nvals = 50) { + #samr.compute.delta.table <- function(samr.obj, + # min.foldchange=0, dels=NULL, nvals=50) { + # computes delta table, starting with samr object 'a', for + # nvals values of delta + lmax = sqrt(max(abs(sort(samr.obj$tt) - samr.obj$evo))) + if (is.null(dels)) { + dels = (seq(0, lmax, length = nvals)^2) + } + col = matrix(1, nrow = length(samr.obj$evo), ncol = nvals) + ttstar0 <- samr.obj$ttstar0 + tt <- samr.obj$tt + n <- samr.obj$n + evo <- samr.obj$evo + nsim <- ncol(ttstar0) + res1 <- NULL + foldchange.cond.up = matrix(T, nrow = nrow(samr.obj$ttstar), + ncol = ncol(samr.obj$ttstar)) + foldchange.cond.lo = matrix(T, nrow = nrow(samr.obj$ttstar), + ncol = ncol(samr.obj$ttstar)) + if (!is.null(samr.obj$foldchange[1]) & (min.foldchange > + 0)) { + foldchange.cond.up = samr.obj$foldchange.star >= min.foldchange + foldchange.cond.lo = samr.obj$foldchange.star <= 1/min.foldchange + } + cutup = rep(NA, length(dels)) + cutlow = rep(NA, length(dels)) + g2 = rep(NA, length(dels)) + errup = matrix(NA, ncol = length(dels), nrow = ncol(samr.obj$ttstar0)) + errlow = matrix(NA, ncol = length(dels), nrow = ncol(samr.obj$ttstar0)) + cat("", fill = T) + cat("Computing delta table", fill = T) + for (ii in 1:length(dels)) { + cat(ii, fill = TRUE) + ttt <- detec.slab(samr.obj, dels[ii], min.foldchange) + cutup[ii] <- 1e+10 + if (length(ttt$pup > 0)) { + cutup[ii] <- min(samr.obj$tt[ttt$pup]) + } + cutlow[ii] <- -1e+10 + if (length(ttt$plow) > 0) { + cutlow[ii] <- max(samr.obj$tt[ttt$plow]) + } + g2[ii] = sumlengths(ttt) + errup[, ii] = colSums(samr.obj$ttstar0 > cutup[ii] & + foldchange.cond.up) + errlow[, ii] = colSums(samr.obj$ttstar0 < cutlow[ii] & + foldchange.cond.lo) + } + s <- sqrt(apply(errup, 2, var)/nsim + apply(errlow, 2, var)/nsim) + gmed <- apply(errup + errlow, 2, median) + g90 = apply(errup + errlow, 2, quantile, 0.9) + res1 <- cbind(samr.obj$pi0 * gmed, samr.obj$pi0 * g90, g2, + samr.obj$pi0 * gmed/g2, samr.obj$pi0 * g90/g2, cutlow, + cutup) + res1 <- cbind(dels, res1) + # remove rows with #called=0 + #om=res1[,4]==0 + #res1=res1[!om,,drop=F] + # remove duplicate rows with same # of genes called + #omm=!duplicated(res1[,4]) + #res1=res1[omm,,drop=F] + dimnames(res1) <- list(NULL, c("delta", "# med false pos", + "90th perc false pos", "# called", "median FDR", "90th perc FDR", + "cutlo", "cuthi")) + return(res1) +} + +####################################################################### +#\tcompute the delta table for sequencing data +####################################################################### +samr.compute.delta.table.seq <- function(samr.obj, + min.foldchange = 0, dels = NULL) { + res1 <- NULL + flag <- T + ## check whether any gene satisfies the foldchange + # restrictions + if ((samr.obj$resp.type == samr.const.twoclass.unpaired.response | + samr.obj$resp.type == samr.const.twoclass.paired.response) & + (min.foldchange > 0)) { + sat.up <- (samr.obj$foldchange >= min.foldchange) & (samr.obj$evo > + 0) + sat.dn <- (samr.obj$foldchange <= 1/min.foldchange) & + (samr.obj$evo < 0) + if (sum(sat.up) + sum(sat.dn) == 0) { + flag <- F + } + } + if (flag) { + if (is.null(dels)) { + dels <- generate.dels(samr.obj, min.foldchange = min.foldchange) + } + cat("Number of thresholds chosen (all possible thresholds) =", + length(dels), fill = T) + if (length(dels) > 0) { + ## sort delta to make the fast calculation right + dels <- sort(dels) + ## get the upper and lower cutoffs + cat("Getting all the cutoffs for the thresholds...\n") + slabs <- samr.seq.detec.slabs(samr.obj, dels, min.foldchange) + cutup <- slabs$cutup + cutlow <- slabs$cutlow + g2 <- slabs$g2 + ## get the number of errors under the null hypothesis + cat("Getting number of false positives in the permutation...\n") + errnum <- samr.seq.null.err(samr.obj, min.foldchange, + cutup, cutlow) + res1 <- NULL + gmed <- apply(errnum, 2, median) + g90 = apply(errnum, 2, quantile, 0.9) + res1 <- cbind(samr.obj$pi0 * gmed, samr.obj$pi0 * + g90, g2, samr.obj$pi0 * gmed/g2, samr.obj$pi0 * + g90/g2, cutlow, cutup) + res1 <- cbind(dels, res1) + dimnames(res1) <- list(NULL, c("delta", "# med false pos", + "90th perc false pos", "# called", "median FDR", + "90th perc FDR", "cutlo", "cuthi")) + } + } + return(res1) +} + +# ============================================================================== +# samr.plot +# ============================================================================== +samr.plot <- function(samr.obj, del = NULL, min.foldchange = 0) { + ## make observed-expected plot + ## takes foldchange into account too + if (is.null(del)) { + del = sqrt(max(abs(sort(samr.obj$tt) - samr.obj$evo))) + } + LARGE = 1e+10 + b <- detec.slab(samr.obj, del, min.foldchange) + bb <- c(b$pup, b$plow) + b1 = LARGE + b0 = -LARGE + if (!is.null(b$pup)) { + b1 <- min(samr.obj$tt[b$pup]) + } + if (!is.null(b$plow)) { + b0 <- max(samr.obj$tt[b$plow]) + } + c1 <- (1:samr.obj$n)[sort(samr.obj$tt) >= b1] + c0 <- (1:samr.obj$n)[sort(samr.obj$tt) <= b0] + c2 <- c(c0, c1) + foldchange.cond.up = rep(T, length(samr.obj$evo)) + foldchange.cond.lo = rep(T, length(samr.obj$evo)) + if (!is.null(samr.obj$foldchange[1]) & (min.foldchange > + 0)) { + foldchange.cond.up = samr.obj$foldchange >= min.foldchange + foldchange.cond.lo = samr.obj$foldchange <= 1/min.foldchange + } + col = rep(1, length(samr.obj$evo)) + col[b$plow] = 3 + col[b$pup] = 2 + if (!is.null(samr.obj$foldchange[1]) & (min.foldchange > + 0)) { + col[!foldchange.cond.lo & !foldchange.cond.up] = 1 + } + col.ordered = col[order(samr.obj$tt)] + ylims <- range(samr.obj$tt) + xlims <- range(samr.obj$evo) + plot(samr.obj$evo, sort(samr.obj$tt), xlab = "expected score", + ylab = "observed score", ylim = ylims, xlim = xlims, + type = "n") + points(samr.obj$evo, sort(samr.obj$tt), col = col.ordered) + abline(0, 1) + abline(del, 1, lty = 2) + abline(-del, 1, lty = 2) +} + +# ============================================================================== +# samr.compute.siggenes.table +# ============================================================================== +samr.compute.siggenes.table = function(samr.obj, del, + data, delta.table, min.foldchange = 0, all.genes = FALSE, + compute.localfdr = FALSE) +{ + ## computes significant genes table, starting with samr + # object 'a' and 'delta.table' + ## for a **single** value del + ## if all.genes is true, all genes are printed (and value + # of del is ignored) + if (is.null(data$geneid)) + { + data$geneid = paste("g", 1:nrow(data$x), sep = "") + } + if (is.null(data$genenames)) + { + data$genenames = paste("g", 1:nrow(data$x), sep = "") + } + if (!all.genes) + { + sig = detec.slab(samr.obj, del, min.foldchange) + } + if (all.genes) + { + p = length(samr.obj$tt) + pup = (1:p)[samr.obj$tt >= 0] + plo = (1:p)[samr.obj$tt < 0] + sig = list(pup = pup, plo = plo) + } + if (compute.localfdr) + { + aa = localfdr(samr.obj, min.foldchange) + if (length(sig$pup) > 0) + { + fdr.up = predictlocalfdr(aa$smooth.object, samr.obj$tt[sig$pup]) + } + if (length(sig$plo) > 0) + { + fdr.lo = predictlocalfdr(aa$smooth.object, samr.obj$tt[sig$plo]) + } + } + qvalues = NULL + if (length(sig$pup) > 0 | length(sig$plo) > 0) + { + qvalues = qvalue.func(samr.obj, sig, delta.table) + } + res.up = NULL + res.lo = NULL + done = FALSE + + # two class unpaired or paired (foldchange is reported) + if ((samr.obj$resp.type == samr.const.twoclass.unpaired.response | + samr.obj$resp.type == samr.const.twoclass.paired.response)) + { + if (!is.null(sig$pup)) + { + res.up = cbind(sig$pup + 1, data$genenames[sig$pup], + data$geneid[sig$pup], samr.obj$tt[sig$pup], samr.obj$numer[sig$pup], + samr.obj$sd[sig$pup], samr.obj$foldchange[sig$pup], + qvalues$qvalue.up) + if (compute.localfdr) + { + res.up = cbind(res.up, fdr.up) + } + temp.names = list(NULL, c("Row", "Gene ID", "Gene Name", + "Score(d)", "Numerator(r)", "Denominator(s+s0)", + "Fold Change", "q-value(%)")) + if (compute.localfdr) + { + temp.names[[2]] = c(temp.names[[2]], "localfdr(%)") + } + dimnames(res.up) = temp.names + } + if (!is.null(sig$plo)) + { + res.lo = cbind(sig$plo + 1, data$genenames[sig$plo], + data$geneid[sig$plo], samr.obj$tt[sig$plo], samr.obj$numer[sig$plo], + samr.obj$sd[sig$plo], samr.obj$foldchange[sig$plo], + qvalues$qvalue.lo) + if (compute.localfdr) + { + res.lo = cbind(res.lo, fdr.lo) + } + temp.names = list(NULL, c("Row", "Gene ID", "Gene Name", + "Score(d)", "Numerator(r)", "Denominator(s+s0)", + "Fold Change", "q-value(%)")) + if (compute.localfdr) + { + temp.names[[2]] = c(temp.names[[2]], "localfdr(%)") + } + dimnames(res.lo) = temp.names + } + done = TRUE + } + + # multiclass + if (samr.obj$resp.type == samr.const.multiclass.response) + { + if (!is.null(sig$pup)) + { + res.up = cbind(sig$pup + 1, data$genenames[sig$pup], + data$geneid[sig$pup], samr.obj$tt[sig$pup], samr.obj$numer[sig$pup], + samr.obj$sd[sig$pup], samr.obj$stand.contrasts[sig$pup, ], qvalues$qvalue.up) + + if (compute.localfdr) + { + res.up = cbind(res.up, fdr.up) + } + + collabs.contrast = paste("contrast-", as.character(1:ncol(samr.obj$stand.contrasts)), + sep = "") + temp.names = list(NULL, c("Row", "Gene ID", "Gene Name", + "Score(d)", "Numerator(r)", "Denominator(s+s0)", + collabs.contrast, "q-value(%)")) + + if (compute.localfdr) + { + temp.names[[2]] = c(temp.names[[2]], "localfdr(%)") + } + dimnames(res.up) = temp.names + } + res.lo = NULL + done = TRUE + } + + #all other cases + if (!done) + { + if (!is.null(sig$pup)) + { + res.up = cbind(sig$pup + 1, data$genenames[sig$pup], + data$geneid[sig$pup], samr.obj$tt[sig$pup], samr.obj$numer[sig$pup], + samr.obj$sd[sig$pup], samr.obj$foldchange[sig$pup], + qvalues$qvalue.up) + if (compute.localfdr) + { + res.up = cbind(res.up, fdr.up) + } + temp.names = list(NULL, c("Row", "Gene ID", "Gene Name", + "Score(d)", "Numerator(r)", "Denominator(s+s0)", + "q-value(%)")) + if (compute.localfdr) + { + temp.names[[2]] = c(temp.names[[2]], "localfdr(%)") + } + dimnames(res.up) = temp.names + } + if (!is.null(sig$plo)) + { + res.lo = cbind(sig$plo + 1, data$genenames[sig$plo], + data$geneid[sig$plo], samr.obj$tt[sig$plo], samr.obj$numer[sig$plo], + samr.obj$sd[sig$plo], samr.obj$foldchange[sig$plo], + qvalues$qvalue.lo) + if (compute.localfdr) + { + res.lo = cbind(res.lo, fdr.lo) + } + temp.names = list(NULL, c("Row", "Gene ID", "Gene Name", + "Score(d)", "Numerator(r)", "Denominator(s+s0)", + "q-value(%)")) + if (compute.localfdr) + { + temp.names[[2]] = c(temp.names[[2]], "localfdr(%)") + } + dimnames(res.lo) = temp.names + } + done = TRUE + } + if (!is.null(res.up)) + { + o1 = order(-samr.obj$tt[sig$pup]) + res.up = res.up[o1, , drop = F] + } + if (!is.null(res.lo)) + { + o2 = order(samr.obj$tt[sig$plo]) + res.lo = res.lo[o2, , drop = F] + } + color.ind.for.multi = NULL + if (samr.obj$resp.type == samr.const.multiclass.response & !is.null(sig$pup)) + { + color.ind.for.multi = 1 * (samr.obj$stand.contrasts[sig$pup, + ] > samr.obj$stand.contrasts.95[2]) + (-1) * (samr.obj$stand.contrasts[sig$pup, + ] < samr.obj$stand.contrasts.95[1]) + } + ngenes.up = nrow(res.up) + if (is.null(ngenes.up)) + { + ngenes.up = 0 + } + ngenes.lo = nrow(res.lo) + if (is.null(ngenes.lo)) + { + ngenes.lo = 0 + } + return(list(genes.up = res.up, genes.lo = res.lo, color.ind.for.multi = color.ind.for.multi, + ngenes.up = ngenes.up, ngenes.lo = ngenes.lo)) +} +generate.dels <- function(samr.obj, min.foldchange = 0) { + dels <- NULL + ## initialize calculation + tag <- order(samr.obj$tt) + if ((samr.obj$resp.type == samr.const.twoclass.unpaired.response | + samr.obj$resp.type == samr.const.twoclass.paired.response) & + (min.foldchange > 0)) { + res.mat <- data.frame(tt = samr.obj$tt[tag], fc = samr.obj$foldchange[tag], + evo = samr.obj$evo, dif = samr.obj$tt[tag] - samr.obj$evo) + res.up <- res.mat[res.mat$evo > 0, ] + res.lo <- res.mat[res.mat$evo < 0, ] + res.up <- res.up[res.up$fc >= min.foldchange, ] + res.lo <- res.lo[res.lo$fc <= 1/min.foldchange, ] + } + else { + res.mat <- data.frame(tt = samr.obj$tt[tag], evo = samr.obj$evo, + dif = samr.obj$tt[tag] - samr.obj$evo) + res.up <- res.mat[res.mat$evo > 0, ] + res.lo <- res.mat[res.mat$evo < 0, ] + } + ## for the upper part + up.vec <- rep(NA, nrow(res.up)) + if (nrow(res.up) > 0) { + st <- 1e-08 + i.cur <- 1 + for (i in 1:nrow(res.up)) { + if (res.up$dif[i] > st) { + st <- res.up$dif[i] + up.vec[i.cur] <- st + i.cur <- i.cur + 1 + } + } + } + ## for the lower part + lo.vec <- rep(NA, nrow(res.lo)) + if (nrow(res.lo) > 0) { + st <- -1e-08 + i.cur <- 1 + for (i in nrow(res.lo):1) { + if (res.lo$dif[i] < st) { + st <- res.lo$dif[i] + lo.vec[i.cur] <- st + i.cur <- i.cur + 1 + } + } + } + ## combine them + vec <- c(up.vec, -lo.vec) + vec <- vec[!is.na(vec)] + vec <- vec - 1e-08 + dels <- sort(unique(vec)) + return(dels) +} +samr.seq.detec.slabs <- function(samr.obj, dels, min.foldchange) { + ## initialize calculation + tag <- order(samr.obj$tt) + if ((samr.obj$resp.type == samr.const.twoclass.unpaired.response | + samr.obj$resp.type == samr.const.twoclass.paired.response) & + (min.foldchange > 0)) { + res.mat <- data.frame(tt = samr.obj$tt[tag], fc = samr.obj$foldchange[tag], + evo = samr.obj$evo, dif = samr.obj$tt[tag] - samr.obj$evo) + res.up <- res.mat[res.mat$evo > 0, ] + res.lo <- res.mat[res.mat$evo < 0, ] + res.up <- res.up[res.up$fc >= min.foldchange, ] + res.lo <- res.lo[res.lo$fc <= 1/min.foldchange, ] + } + else { + res.mat <- data.frame(tt = samr.obj$tt[tag], evo = samr.obj$evo, + dif = samr.obj$tt[tag] - samr.obj$evo) + res.up <- res.mat[res.mat$evo > 0, ] + res.lo <- res.mat[res.mat$evo < 0, ] + } + ## begin calculating + cutup <- rep(1e+10, length(dels)) + cutlow <- rep(-1e+10, length(dels)) + g2.up <- g2.lo <- rep(0, length(dels)) + if (nrow(res.up) > 0) { + res.up <- data.frame(dif = res.up$dif, tt = res.up$tt, + num = nrow(res.up):1) + ## get the upper part + j <- 1 + ii <- 1 + while (j <= nrow(res.up) & ii <= length(dels)) { + if (res.up$dif[j] > dels[ii]) { + cutup[ii] <- res.up$tt[j] + g2.up[ii] <- res.up$num[j] + ii <- ii + 1 + } + else { + j <- j + 1 + } + } + } + if (nrow(res.lo) > 0) { + res.lo <- data.frame(dif = res.lo$dif, tt = res.lo$tt, + num = 1:nrow(res.lo)) + ## get the lower part + j <- nrow(res.lo) + ii <- 1 + while (j >= 1 & ii <= length(dels)) { + if (res.lo$dif[j] < -dels[ii]) { + cutlow[ii] <- res.lo$tt[j] + g2.lo[ii] <- res.lo$num[j] + ii <- ii + 1 + } + else { + j <- j - 1 + } + } + } + g2 <- g2.up + g2.lo + return(list(cutup = cutup, cutlow = cutlow, g2 = g2)) +} +sumlengths <- function(aa) { + length(aa$pl) + length(aa$pu) +} + +samr.seq.null.err <- function(samr.obj, min.foldchange, + cutup, cutlow) { + errup = matrix(NA, ncol = length(cutup), nrow = ncol(samr.obj$ttstar0)) + errlow = matrix(NA, ncol = length(cutlow), nrow = ncol(samr.obj$ttstar0)) + cutup.rank <- rank(cutup, ties.method = "min") + cutlow.rank <- rank(-cutlow, ties.method = "min") + for (jj in 1:ncol(samr.obj$ttstar0)) { + #cat(jj, fill=TRUE) + keep.up <- keep.dn <- samr.obj$ttstar0[, jj] + if ((samr.obj$resp.type == samr.const.twoclass.unpaired.response | + samr.obj$resp.type == samr.const.twoclass.paired.response) & + (min.foldchange > 0)) { + keep.up <- keep.up[samr.obj$foldchange.star[, jj] >= + min.foldchange] + keep.dn <- keep.dn[samr.obj$foldchange.star[, jj] <= + 1/min.foldchange] + } + errup[jj, ] <- length(keep.up) - (rank(c(cutup, keep.up), + ties.method = "min")[1:length(cutup)] - cutup.rank) + errlow[jj, ] <- length(keep.dn) - (rank(c(-cutlow, -keep.dn), + ties.method = "min")[1:length(cutlow)] - cutlow.rank) + } + errnum <- errup + errlow + return(errnum) +} +detec.slab <- function(samr.obj, del, min.foldchange) { + ## find genes above and below the slab of half-width del + # this calculation is tricky- for consistency, the slab + # condition picks + # all genes that are beyond the first departure from the + # slab + # then the fold change condition is applied (if applicable) + n <- length(samr.obj$tt) + tt <- samr.obj$tt + evo <- samr.obj$evo + numer <- samr.obj$tt * (samr.obj$sd + samr.obj$s0) + tag <- order(tt) + pup <- NULL + foldchange.cond.up = rep(T, length(evo)) + foldchange.cond.lo = rep(T, length(evo)) + if (!is.null(samr.obj$foldchange[1]) & (min.foldchange > + 0)) { + foldchange.cond.up = samr.obj$foldchange >= min.foldchange + foldchange.cond.lo = samr.obj$foldchange <= 1/min.foldchange + } + o1 <- (1:n)[(tt[tag] - evo > del) & evo > 0] + if (length(o1) > 0) { + o1 <- o1[1] + o11 <- o1:n + o111 <- rep(F, n) + o111[tag][o11] <- T + pup <- (1:n)[o111 & foldchange.cond.up] + } + plow <- NULL + o2 <- (1:n)[(evo - tt[tag] > del) & evo < 0] + if (length(o2) > 0) { + o2 <- o2[length(o2)] + o22 <- 1:o2 + o222 <- rep(F, n) + o222[tag][o22] <- T + plow <- (1:n)[o222 & foldchange.cond.lo] + } + return(list(plow = plow, pup = pup)) +} + +#' @importFrom stats smooth.spline +localfdr <- function(samr.obj, min.foldchange, perc = 0.01, + df = 10) { + ## estimates compute.localfdr at score 'd', using SAM + # object 'samr.obj' + ## 'd' can be a vector of d scores + ## returns estimate of symmetric fdr as a percentage + # this version uses a 1% symmetric window, and does not + # estimate fdr in + # windows having fewer than 100 genes + ## to use: first run samr and then pass the resulting fit + # object to + ## localfdr + ## NOTE: at most 20 of the perms are used to estimate the + # fdr (for speed sake) + # I try two window shapes: symmetric and an assymetric one + # currently I use the symmetric window to estimate the + # compute.localfdr + ngenes = length(samr.obj$tt) + mingenes = 50 + # perc is increased, in order to get at least mingenes in a + # window + perc = max(perc, mingenes/length(samr.obj$tt)) + nperms.to.use = min(20, ncol(samr.obj$ttstar)) + nperms = ncol(samr.obj$ttstar) + d = seq(sort(samr.obj$tt)[1], sort(samr.obj$tt)[ngenes], + length = 100) + ndscore <- length(d) + dvector <- rep(NA, ndscore) + ind.foldchange = rep(T, length(samr.obj$tt)) + if (!is.null(samr.obj$foldchange[1]) & min.foldchange > 0) { + ind.foldchange = (samr.obj$foldchange >= min.foldchange) | + (samr.obj$foldchange <= min.foldchange) + } + fdr.temp = function(temp, dlow, dup, pi0, ind.foldchange) { + return(sum(pi0 * (temp >= dlow & temp <= dup & ind.foldchange))) + } + for (i in 1:ndscore) { + pi0 <- samr.obj$pi0 + r <- sum(samr.obj$tt < d[i]) + r22 <- round(max(r - length(samr.obj$tt) * perc/2, 1)) + dlow.sym <- sort(samr.obj$tt)[r22] + # if(d[i]<0) + # { + # r2 <- max(r-length(samr.obj$tt)*perc/2, 1) + # r22= min(r+length(samr.obj$tt)*perc/2, + # length(samr.obj$tt)) + # + # dlow <- sort(samr.obj$tt)[r2] + # dup=sort(samr.obj$tt)[r22] + # } + r22 <- min(r + length(samr.obj$tt) * perc/2, length(samr.obj$tt)) + dup.sym <- sort(samr.obj$tt)[r22] + # if(d[i]>0) + # { + # r2 <- min(r+length(samr.obj$tt)*perc/2, + # length(samr.obj$tt)) + # r22 <- max(r-length(samr.obj$tt)*perc/2, 1) + # dup <- sort(samr.obj$tt)[r2] + # dlow <- sort(samr.obj$tt)[r22] + # + # } + # o <- samr.obj$tt>=dlow & samr.obj$tt<= dup & + # ind.foldchange + oo <- samr.obj$tt >= dlow.sym & samr.obj$tt <= dup.sym & + ind.foldchange + nsim <- ncol(samr.obj$ttstar) + fdr <- rep(NA, nsim) + fdr2 <- fdr + if (!is.null(samr.obj$foldchange[1]) & min.foldchange > + 0) { + temp = as.vector(samr.obj$foldchange.star[, 1:nperms.to.use]) + ind.foldchange = (temp >= min.foldchange) | (temp <= + min.foldchange) + } + temp = samr.obj$ttstar0[, sample(1:nperms, size = nperms.to.use)] + # fdr <-median(apply(temp,2,fdr.temp,dlow, dup, pi0, + # ind.foldchange)) + fdr.sym <- median(apply(temp, 2, fdr.temp, dlow.sym, + dup.sym, pi0, ind.foldchange)) + # fdr <- 100*fdr/sum(o) + fdr.sym <- 100 * fdr.sym/sum(oo) + dlow.sym <- dlow.sym + dup.sym <- dup.sym + dvector[i] <- fdr.sym + } + om = !is.na(dvector) & (dvector != Inf) + aa = smooth.spline(d[om], dvector[om], df = df) + return(list(smooth.object = aa, perc = perc, df = df)) +} + +predictlocalfdr = function(smooth.object, d) { + yhat = predict(smooth.object, d)$y + yhat = pmin(yhat, 100) + yhat = pmax(yhat, 0) + return(yhat) +} + +qvalue.func = function(samr.obj, sig, delta.table) { + # returns q-value as a percentage (out of 100) + LARGE = 1e+10 + qvalue.up = rep(NA, length(sig$pup)) + o1 = sig$pup + cutup = delta.table[, 8] + FDR = delta.table[, 5] + ii = 0 + for (i in o1) { + o = abs(cutup - samr.obj$tt[i]) + o[is.na(o)] = LARGE + oo = (1:length(o))[o == min(o)] + oo = oo[length(oo)] + ii = ii + 1 + qvalue.up[ii] = FDR[oo] + } + qvalue.lo = rep(NA, length(sig$plo)) + o2 = sig$plo + cutlo = delta.table[, 7] + ii = 0 + for (i in o2) { + o = abs(cutlo - samr.obj$tt[i]) + o[is.na(o)] = LARGE + oo = (1:length(o))[o == min(o)] + oo = oo[length(oo)] + ii = ii + 1 + qvalue.lo[ii] = FDR[oo] + } + # any qvalues that are missing, are set to 1 (the highest + # value) + qvalue.lo[is.na(qvalue.lo)] = 1 + qvalue.up[is.na(qvalue.up)] = 1 + # ensure that each qvalue vector is monotone non-increasing + o1 = order(samr.obj$tt[sig$plo]) + qv1 = qvalue.lo[o1] + qv11 = qv1 + if (length(qv1) > 1) { + for (i in 2:length(qv1)) { + if (qv11[i] < qv11[i - 1]) { + qv11[i] = qv11[i - 1] + } + } + qv111 = qv11 + qv111[o1] = qv11 + } + else { + qv111 = qv1 + } + o2 = order(samr.obj$tt[sig$pup]) + qv2 = qvalue.up[o2] + qv22 = qv2 + if (length(qv2) > 1) { + for (i in 2:length(qv2)) { + if (qv22[i] > qv22[i - 1]) { + qv22[i] = qv22[i - 1] + } + } + qv222 = qv22 + qv222[o2] = qv22 + } + else { + qv222 = qv2 + } + return(list(qvalue.lo = 100 * qv111, qvalue.up = 100 * qv222)) +} From da2f7b4e2f77715a6aac256e6cb0772010115be5 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 3 Aug 2020 14:46:37 +0200 Subject: [PATCH 75/99] Removed seed setting on FindOutliersMB --- R/DIscBIO-generic-FindOutliersMB.R | 1 - 1 file changed, 1 deletion(-) diff --git a/R/DIscBIO-generic-FindOutliersMB.R b/R/DIscBIO-generic-FindOutliersMB.R index f3116f7..3a20cb1 100644 --- a/R/DIscBIO-generic-FindOutliersMB.R +++ b/R/DIscBIO-generic-FindOutliersMB.R @@ -244,7 +244,6 @@ setMethod( object@cpart <- cpart - set.seed(111111) object@fcol <- sample(rainbow(max(cpart))) p <- object@MBclusters$clusterid[order( object@MBclusters$clusterid, decreasing = FALSE From f27907a9b97116de5a7bd24fd36a12b933cbc883 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 3 Aug 2020 14:54:02 +0200 Subject: [PATCH 76/99] Merged FindOutliersMB into FindOutliers --- DESCRIPTION | 3 +- NAMESPACE | 3 +- ...ersKM.R => DIscBIO-generic-FindOutliers.R} | 57 +++- R/DIscBIO-generic-FindOutliersMB.R | 297 ------------------ man/{FindOutliersKM.Rd => FindOutliers.Rd} | 18 +- man/FindOutliersMB.Rd | 66 ---- 6 files changed, 51 insertions(+), 393 deletions(-) rename R/{DIscBIO-generic-FindOutliersKM.R => DIscBIO-generic-FindOutliers.R} (88%) delete mode 100644 R/DIscBIO-generic-FindOutliersMB.R rename man/{FindOutliersKM.Rd => FindOutliers.Rd} (81%) delete mode 100644 man/FindOutliersMB.Rd diff --git a/DESCRIPTION b/DESCRIPTION index a7162d4..2c2b3cc 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -62,8 +62,7 @@ Collate: 'DIscBIO-generic-DEGanalysis2clust.R' 'DIscBIO-generic-Exprmclust.R' 'DIscBIO-generic-FinalPreprocessing.R' - 'DIscBIO-generic-FindOutliersKM.R' - 'DIscBIO-generic-FindOutliersMB.R' + 'DIscBIO-generic-FindOutliers.R' 'DIscBIO-generic-KMClustDiffGenes.R' 'DIscBIO-generic-KMclustheatmap.R' 'DIscBIO-generic-KmeanOrder.R' diff --git a/NAMESPACE b/NAMESPACE index 2ce6381..8a3fd94 100644 --- a/NAMESPACE +++ b/NAMESPACE @@ -25,8 +25,7 @@ exportMethods(DEGanalysis) exportMethods(DEGanalysis2clust) exportMethods(Exprmclust) exportMethods(FinalPreprocessing) -exportMethods(FindOutliersKM) -exportMethods(FindOutliersMB) +exportMethods(FindOutliers) exportMethods(KMClustDiffGenes) exportMethods(KMclustheatmap) exportMethods(KmeanOrder) diff --git a/R/DIscBIO-generic-FindOutliersKM.R b/R/DIscBIO-generic-FindOutliers.R similarity index 88% rename from R/DIscBIO-generic-FindOutliersKM.R rename to R/DIscBIO-generic-FindOutliers.R index aa11009..131fd13 100644 --- a/R/DIscBIO-generic-FindOutliersKM.R +++ b/R/DIscBIO-generic-FindOutliers.R @@ -1,5 +1,5 @@ -#' @title Inference of outlier cells in K-means clustering -#' @description This functions performs the outlier identification +#' @title Inference of outlier cells +#' @description This functions performs the outlier identification for k-means and model-based clustering #' @param object \code{DISCBIO} class object. #' @param outminc minimal transcript count of a gene in a clusters to be tested #' for being an outlier gene. Default is 5. @@ -10,7 +10,7 @@ #' binomial background model of expression in a cluster. Default is 0.001. #' @param thr probability values for which the number of outliers is computed in #' order to plot the dependence of the number of outliers on the probability -#' threshold. Default is 2**-(1:40). +#' threshold. Default is 2**-(1:40).set #' @param outdistquant Real number between zero and one. Outlier cells are #' merged to outlier clusters if their distance smaller than the #' outdistquant-quantile of the distance distribution of pairs of cells in @@ -23,25 +23,48 @@ #' of cells on each. #' setGeneric( - "FindOutliersKM", + "FindOutliers", function( object, K, outminc = 5, outlg = 2, probthr = 1e-3, thr = 2 ** -(1:40), outdistquant = .75, plot = TRUE, quiet = FALSE ) - standardGeneric("FindOutliersKM") + standardGeneric("FindOutliers") ) -#' @rdname FindOutliersKM +#' @rdname FindOutliers #' @export setMethod( - "FindOutliersKM", + "FindOutliers", signature = "DISCBIO", definition = function( object, K, outminc, outlg, probthr, thr, outdistquant, plot, quiet ) { - if (length(object@kmeans$kpart) == 0) - stop("run Clustexp before FindOutliersKM") + # ====================================================================== + # Validating + # ====================================================================== + ran_k <- length(object@kmeans$kpart) > 0 + ran_m <- length(object@MBclusters$clusterid) > 0 + if (ran_k) { + clusters <- object@kmeans$kpart + } else if (ran_m) { + object <- Clustexp( + object, + clustnr = 20, + bootnr = 50, + metric = "pearson", + do.gap = T, + SE.method = "Tibs2001SEmax", + SE.factor = .25, + B.gap = 50, + cln = K, + rseed = 17000, + quiet = quiet + ) + clusters <- object@MBclusters$clusterid + } else { + stop("run Clustexp before FindOutliers") + } if (!is.numeric(outminc)) stop("outminc has to be a non-negative integer") else if (round(outminc) != outminc | outminc < 0) @@ -105,15 +128,15 @@ setMethod( out <- c() stest <- rep(0, length(thr)) cprobs <- c() - for (n in 1:max(object@kmeans$kpart)) { - if (sum(object@kmeans$kpart == n) == 1) { + for (n in 1:max(clusters)) { + if (sum(clusters == n) == 1) { cprobs <- append(cprobs, .5) names(cprobs)[length(cprobs)] <- - names(object@kmeans$kpart)[object@kmeans$kpart == n] + names(clusters)[clusters == n] next } - x <- object@fdata[, object@kmeans$kpart == n] + x <- object@fdata[, clusters == n] x <- x[apply(x, 1, max) > outminc, ] z <- t(apply(x, 1, function(x) { @@ -156,8 +179,8 @@ setMethod( clp2p.cl <- c() cols <- names(object@fdata) di <- as.data.frame(object@distances) - for (i in 1:max(object@kmeans$kpart)) { - tcol <- cols[object@kmeans$kpart == i] + for (i in 1:max(clusters)) { + tcol <- cols[clusters == i] if (sum(!(tcol %in% out)) > 1) clp2p.cl <- append( clp2p.cl, @@ -168,7 +191,7 @@ setMethod( } clp2p.cl <- clp2p.cl[clp2p.cl > 0] - cpart <- object@kmeans$kpart + cpart <- clusters cadd <- list() if (length(out) > 0) { if (length(out) == 1) { @@ -221,7 +244,7 @@ setMethod( object@fcol <- rainbow(max(cpart)) p <- - object@kmeans$kpart[order(object@kmeans$kpart, decreasing = FALSE)] + clusters[order(clusters, decreasing = FALSE)] x <- object@out$cprobs[names(p)] fcol <- c("black", "blue", "green", "red", "yellow", "gray") if (plot) { diff --git a/R/DIscBIO-generic-FindOutliersMB.R b/R/DIscBIO-generic-FindOutliersMB.R deleted file mode 100644 index 3a20cb1..0000000 --- a/R/DIscBIO-generic-FindOutliersMB.R +++ /dev/null @@ -1,297 +0,0 @@ -#' @title Inference of outlier cells in Model-based clustering -#' @description This functions performs the outlier identification -#' @param object \code{DISCBIO} class object. -#' @param outminc minimal transcript count of a gene in a clusters to be tested -#' for being an outlier gene. Default is 5. -#' @param outlg Minimum number of outlier genes required for being an outlier -#' cell. Default is 2. -#' @param probthr outlier probability threshold for a minimum of \code{outlg} -#' genes to be an outlier cell. This probability is computed from a negative -#' binomial background model of expression in a cluster. Default is 0.001. -#' @param thr probability values for which the number of outliers is computed in -#' order to plot the dependence of the number of outliers on the probability -#' threshold. Default is 2**-(1:40). -#' @param outdistquant Real number between zero and one. Outlier cells are -#' merged to outlier clusters if their distance smaller than the -#' outdistquant-quantile of the distance distribution of pairs of cells in -#' the orginal clusters after outlier removal. Default is 0.75. -#' @param K Number of clusters to be used. -#' @param plot if `TRUE`, produces a plot of -log10prob per K -#' @param quiet if `TRUE`, intermediary output is suppressed -#' @importFrom stats coef pnbinom -#' @return A named vector of the genes containing outlying cells and the number -#' of cells on each. -setGeneric( - name = "FindOutliersMB", - def = function(object, - K, - outminc = 5, - outlg = 2, - probthr = 1e-3, - thr = 2 ** -(1:40), - outdistquant = .75, - plot = TRUE, - quiet = FALSE) { - standardGeneric("FindOutliersMB") - } -) - -#' @export -#' @rdname FindOutliersMB -setMethod( - f = "FindOutliersMB", - signature = "DISCBIO", - definition = function(object, - K, - outminc, - outlg, - probthr, - thr, - outdistquant, - plot = TRUE, - quiet = FALSE) { - if (length(object@MBclusters$clusterid) == 0) - stop("run exprmclust before FindOutliersMB") - if (!is.numeric(outminc)) - stop("outminc has to be a non-negative integer") - else if (round(outminc) != outminc | - outminc < 0) - stop("outminc has to be a non-negative integer") - if (!is.numeric(outlg)) - stop("outlg has to be a non-negative integer") - else if (round(outlg) != outlg | - outlg < 0) - stop("outlg has to be a non-negative integer") - if (!is.numeric(probthr)) - stop("probthr has to be a number between 0 and 1") - else if (probthr < 0 | - probthr > 1) - stop("probthr has to be a number between 0 and 1") - if (!is.numeric(thr)) - stop("thr hast to be a vector of numbers between 0 and 1") - else if (min(thr) < 0 | - max(thr) > 1) - stop("thr hast to be a vector of numbers between 0 and 1") - if (!is.numeric(outdistquant)) - stop("outdistquant has to be a number between 0 and 1") - else if (outdistquant < 0 | - outdistquant > 1) - stop("outdistquant has to be a number between 0 and 1") - - object <- Clustexp( - object, - clustnr = 20, - bootnr = 50, - metric = "pearson", - do.gap = T, - SE.method = "Tibs2001SEmax", - SE.factor = .25, - B.gap = 50, - cln = K, - rseed = 17000, - quiet = quiet - ) - - object@outlierpar <- - list( - outminc = outminc, - outlg = outlg, - probthr = probthr, - thr = thr, - outdistquant = outdistquant - ) - - ### calibrate background model - m <- log2(apply(object@fdata, 1, mean)) - v <- log2(apply(object@fdata, 1, var)) - f <- m > -Inf & v > -Inf - m <- m[f] - v <- v[f] - mm <- -8 - repeat { - fit <- lm(v ~ m + I(m ^ 2)) - if (coef(fit)[3] >= 0 | mm >= 3) { - break - } - mm <- mm + .5 - f <- m > mm - m <- m[f] - v <- v[f] - } - object@background <- list() - object@background$vfit <- fit - object@background$lvar <- function(x, object) { - 2 ** ( - coef(object@background$vfit)[1] + - log2(x) * coef(object@background$vfit)[2] + - coef(object@background$vfit)[3] * - log2(x) ** 2 - ) - } - object@background$lsize <- function(x, object) { - x ** 2 / (max(x + 1e-6, object@background$lvar(x, object)) - x) - } - - ### identify outliers - out <- c() - stest <- rep(0, length(thr)) - cprobs <- c() - for (n in 1:max(object@MBclusters$clusterid)) { - if (sum(object@MBclusters$clusterid == n) == 1) { - cprobs <- - append(cprobs, .5) - names(cprobs)[length(cprobs)] <- - names(object@MBclusters$clusterid)[object@MBclusters$clusterid == n] - next - } - x <- object@fdata[, object@MBclusters$clusterid == n] - x <- x[apply(x, 1, max) > outminc, ] - z <- - t(apply(x, 1, function(x) { - apply(cbind( - pnbinom( - round(x, 0), - mu = mean(x), - size = object@background$lsize(mean(x), object) - ) , - 1 - pnbinom( - round(x, 0), - mu = mean(x), - size = object@background$lsize(mean(x), object) - ) - ), 1, min) - })) - cp <- - apply(z, 2, function(x) { - y <- - p.adjust(x, method = "BH") - y <- y[order(y, decreasing = FALSE)] - return(y[outlg]) - }) - f <- cp < probthr - cprobs <- append(cprobs, cp) - if (sum(f) > 0) - out <- append(out, names(x)[f]) - for (j in 1:length(thr)) - stest[j] <- stest[j] + sum(cp < thr[j]) - } - object@out <- list( - out = out, - stest = stest, - thr = thr, - cprobs = cprobs - ) - - ### cluster outliers - clp2p.cl <- c() - cols <- names(object@fdata) - di <- as.data.frame(object@distances) - for (i in 1:max(object@MBclusters$clusterid)) { - tcol <- cols[object@MBclusters$clusterid == i] - if (sum(!(tcol %in% out)) > 1) - clp2p.cl <- append( - clp2p.cl, - as.vector(t(di[tcol[!(tcol %in% out)], tcol[!(tcol %in% out)]])) - ) - } - clp2p.cl <- clp2p.cl[clp2p.cl > 0] - - cpart <- object@MBclusters$clusterid - cadd <- list() - if (length(out) > 0) { - if (length(out) == 1) { - cadd <- list(out) - } else{ - n <- out - m <- as.data.frame(di[out, out]) - - for (i in 1:length(out)) { - if (length(n) > 1) { - o <- - order(apply(cbind(m, 1:dim(m)[1]), 1, function(x) - min(x[1:(length(x) - 1)][-x[length(x)]])), decreasing = FALSE) - m <- m[o, o] - n <- n[o] - f <- m[, 1] < quantile(clp2p.cl, outdistquant) | - m[, 1] == min(clp2p.cl) - ind <- 1 - if (sum(f) > 1) - for (j in 2:sum(f)) - comp1 <- m[f, f][j, c(ind, j)] - comp2 <- quantile(clp2p.cl, outdistquant) - if (apply(comp1 > comp2, 1, sum) == 0) - ind <- append(ind, j) - cadd[[i]] <- n[f][ind] - g <- !n %in% n[f][ind] - n <- n[g] - m <- m[g, g] - if (sum(g) == 0) - break - - } else if (length(n) == 1) { - cadd[[i]] <- n - break - } - } - } - - for (i in 1:length(cadd)) { - cpart[cols %in% cadd[[i]]] <- max(cpart) + 1 - } - } - - ### determine final clusters - - object@cpart <- cpart - - object@fcol <- sample(rainbow(max(cpart))) - p <- object@MBclusters$clusterid[order( - object@MBclusters$clusterid, decreasing = FALSE - )] - x <- object@out$cprobs[names(p)] - fcol <- c("black", "blue", "green", "red", "yellow", "gray") - if (plot) { - for (i in 1:max(p)) { - y <- -log10(x + 2.2e-16) - y[p != i] <- 0 - if (i == 1) - b <- - barplot( - y, - ylim = c(0, max(-log10(x + 2.2e-16)) * 1.1), - col = fcol[i], - border = fcol[i], - names.arg = FALSE, - ylab = "-log10prob" - ) - else - barplot( - y, - add = TRUE, - col = fcol[i], - border = fcol[i], - names.arg = FALSE, - axes = FALSE - ) - } - abline(-log10(object@outlierpar$probthr), - 0, - col = "black", - lty = 2) - d <- b[2, 1] - b[1, 1] - y <- 0 - for (i in 1:max(p)) - y <- append(y, b[sum(p <= i), 1] + d / 2) - axis(1, at = (y[1:(length(y) - 1)] + y[-1]) / 2, labels = 1:max(p)) - box() - } - if (!quiet) { - message("The following cells are considered as outlier cells:", - which(object@cpart > K), - "\n") - print(which(object@cpart > K)) - } - LL = which(object@cpart > K) - return(LL) - } - ) \ No newline at end of file diff --git a/man/FindOutliersKM.Rd b/man/FindOutliers.Rd similarity index 81% rename from man/FindOutliersKM.Rd rename to man/FindOutliers.Rd index fdc8124..2d9913e 100644 --- a/man/FindOutliersKM.Rd +++ b/man/FindOutliers.Rd @@ -1,11 +1,11 @@ % Generated by roxygen2: do not edit by hand -% Please edit documentation in R/DIscBIO-generic-FindOutliersKM.R -\name{FindOutliersKM} -\alias{FindOutliersKM} -\alias{FindOutliersKM,DISCBIO-method} -\title{Inference of outlier cells in K-means clustering} +% Please edit documentation in R/DIscBIO-generic-FindOutliers.R +\name{FindOutliers} +\alias{FindOutliers} +\alias{FindOutliers,DISCBIO-method} +\title{Inference of outlier cells} \usage{ -FindOutliersKM( +FindOutliers( object, K, outminc = 5, @@ -17,7 +17,7 @@ FindOutliersKM( quiet = FALSE ) -\S4method{FindOutliersKM}{DISCBIO}( +\S4method{FindOutliers}{DISCBIO}( object, K, outminc = 5, @@ -46,7 +46,7 @@ binomial background model of expression in a cluster. Default is 0.001.} \item{thr}{probability values for which the number of outliers is computed in order to plot the dependence of the number of outliers on the probability -threshold. Default is 2**-(1:40).} +threshold. Default is 2**-(1:40).set} \item{outdistquant}{Real number between zero and one. Outlier cells are merged to outlier clusters if their distance smaller than the @@ -62,5 +62,5 @@ A named vector of the genes containing outlying cells and the number of cells on each. } \description{ -This functions performs the outlier identification +This functions performs the outlier identification for k-means and model-based clustering } diff --git a/man/FindOutliersMB.Rd b/man/FindOutliersMB.Rd deleted file mode 100644 index 82173b3..0000000 --- a/man/FindOutliersMB.Rd +++ /dev/null @@ -1,66 +0,0 @@ -% Generated by roxygen2: do not edit by hand -% Please edit documentation in R/DIscBIO-generic-FindOutliersMB.R -\name{FindOutliersMB} -\alias{FindOutliersMB} -\alias{FindOutliersMB,DISCBIO-method} -\title{Inference of outlier cells in Model-based clustering} -\usage{ -FindOutliersMB( - object, - K, - outminc = 5, - outlg = 2, - probthr = 0.001, - thr = 2^-(1:40), - outdistquant = 0.75, - plot = TRUE, - quiet = FALSE -) - -\S4method{FindOutliersMB}{DISCBIO}( - object, - K, - outminc = 5, - outlg = 2, - probthr = 0.001, - thr = 2^-(1:40), - outdistquant = 0.75, - plot = TRUE, - quiet = FALSE -) -} -\arguments{ -\item{object}{\code{DISCBIO} class object.} - -\item{K}{Number of clusters to be used.} - -\item{outminc}{minimal transcript count of a gene in a clusters to be tested -for being an outlier gene. Default is 5.} - -\item{outlg}{Minimum number of outlier genes required for being an outlier -cell. Default is 2.} - -\item{probthr}{outlier probability threshold for a minimum of \code{outlg} -genes to be an outlier cell. This probability is computed from a negative -binomial background model of expression in a cluster. Default is 0.001.} - -\item{thr}{probability values for which the number of outliers is computed in -order to plot the dependence of the number of outliers on the probability -threshold. Default is 2**-(1:40).} - -\item{outdistquant}{Real number between zero and one. Outlier cells are -merged to outlier clusters if their distance smaller than the -outdistquant-quantile of the distance distribution of pairs of cells in -the orginal clusters after outlier removal. Default is 0.75.} - -\item{plot}{if `TRUE`, produces a plot of -log10prob per K} - -\item{quiet}{if `TRUE`, intermediary output is suppressed} -} -\value{ -A named vector of the genes containing outlying cells and the number - of cells on each. -} -\description{ -This functions performs the outlier identification -} From 4899bc431af9d1b77f4bc329134c19bdc8dd6cbf Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 3 Aug 2020 15:07:30 +0200 Subject: [PATCH 77/99] Merged KMClustDiffGenes and MBClustDiffGenes --- DESCRIPTION | 3 +- NAMESPACE | 8 +- ...nes.R => DIscBIO-generic-ClustDiffGenes.R} | 34 ++- R/DIscBIO-generic-MBClustDiffGenes.R | 224 ------------------ ...{KMClustDiffGenes.Rd => ClustDiffGenes.Rd} | 12 +- man/MBClustDiffGenes.Rd | 61 ----- tests/testthat/test-DIscBIO.IMP.R | 2 +- 7 files changed, 34 insertions(+), 310 deletions(-) rename R/{DIscBIO-generic-KMClustDiffGenes.R => DIscBIO-generic-ClustDiffGenes.R} (89%) delete mode 100644 R/DIscBIO-generic-MBClustDiffGenes.R rename man/{KMClustDiffGenes.Rd => ClustDiffGenes.Rd} (85%) delete mode 100644 man/MBClustDiffGenes.Rd diff --git a/DESCRIPTION b/DESCRIPTION index 2c2b3cc..8b87352 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -57,16 +57,15 @@ BugReports: https://github.com/ocbe-uio/DIscBIO/issues Collate: 'DIscBIO-classes.R' 'DIscBIO-generic-ClassVectoringDT.R' + 'DIscBIO-generic-ClustDiffGenes.R' 'DIscBIO-generic-Clustexp.R' 'DIscBIO-generic-DEGanalysis.R' 'DIscBIO-generic-DEGanalysis2clust.R' 'DIscBIO-generic-Exprmclust.R' 'DIscBIO-generic-FinalPreprocessing.R' 'DIscBIO-generic-FindOutliers.R' - 'DIscBIO-generic-KMClustDiffGenes.R' 'DIscBIO-generic-KMclustheatmap.R' 'DIscBIO-generic-KmeanOrder.R' - 'DIscBIO-generic-MBClustDiffGenes.R' 'DIscBIO-generic-MBclustheatmap.R' 'DIscBIO-generic-NoiseFiltering.R' 'DIscBIO-generic-Normalizedata.R' diff --git a/NAMESPACE b/NAMESPACE index 8a3fd94..37a8dc8 100644 --- a/NAMESPACE +++ b/NAMESPACE @@ -1,12 +1,11 @@ # Generated by roxygen2: do not edit by hand +export(ClustDiffGenes) export(DISCBIO) export(DISCBIO2SingleCellExperiment) export(J48DT) export(J48DTeval) export(Jaccard) -export(KMClustDiffGenes) -export(MBClustDiffGenes) export(MB_Order) export(NetAnalysis) export(Networking) @@ -20,16 +19,15 @@ export(customConvertFeats) export(plotGap) exportClasses(DISCBIO) exportMethods(ClassVectoringDT) +exportMethods(ClustDiffGenes) exportMethods(Clustexp) exportMethods(DEGanalysis) exportMethods(DEGanalysis2clust) exportMethods(Exprmclust) exportMethods(FinalPreprocessing) exportMethods(FindOutliers) -exportMethods(KMClustDiffGenes) exportMethods(KMclustheatmap) exportMethods(KmeanOrder) -exportMethods(MBClustDiffGenes) exportMethods(MBclustheatmap) exportMethods(NoiseFiltering) exportMethods(Normalizedata) @@ -44,10 +42,8 @@ exportMethods(plotSymbolstSNE) exportMethods(plotexptsneMB) exportMethods(plottSNE) exportMethods(plottsneMB) -import(org.Hs.eg.db) importClassesFrom(SingleCellExperiment,SingleCellExperiment) importFrom(AnnotationDbi,keys) -importFrom(AnnotationDbi,select) importFrom(NetIndices,GenInd) importFrom(RColorBrewer,brewer.pal) importFrom(RWeka,J48) diff --git a/R/DIscBIO-generic-KMClustDiffGenes.R b/R/DIscBIO-generic-ClustDiffGenes.R similarity index 89% rename from R/DIscBIO-generic-KMClustDiffGenes.R rename to R/DIscBIO-generic-ClustDiffGenes.R index a0bcad4..a178d08 100644 --- a/R/DIscBIO-generic-KMClustDiffGenes.R +++ b/R/DIscBIO-generic-ClustDiffGenes.R @@ -12,12 +12,12 @@ #' @param filename_binom Name of the exported binomial file #' @param filename_sigdeg Name of the exported sigDEG file #' @importFrom stats pbinom median -#' @rdname KMClustDiffGenes +#' @rdname ClustDiffGenes #' @return A list containing two tables. #' @export #' setGeneric( - "KMClustDiffGenes", + "ClustDiffGenes", function( object, K, pValue = 0.05, fdr = .01, export = FALSE, quiet = FALSE, filename_up = "Up-DEG-cluster", @@ -25,19 +25,24 @@ setGeneric( filename_binom = "binomial-DEGsTable", filename_sigdeg = "binomial-sigDEG" ) { - standardGeneric("KMClustDiffGenes") + standardGeneric("ClustDiffGenes") } ) #' @export -#' @rdname KMClustDiffGenes +#' @rdname ClustDiffGenes setMethod( - "KMClustDiffGenes", + "ClustDiffGenes", signature = "DISCBIO", definition = function( object, K, pValue, fdr, export, quiet, filename_up, filename_down, filename_binom, filename_sigdeg - ) { - # Validation + ) + { + # ====================================================================== + # Validating + # ====================================================================== + ran_k <- length(object@kmeans$kpart) > 0 + ran_m <- length(object@MBclusters) > 0 if (!is.numeric(fdr)) { stop("fdr has to be a number between 0 and 1") } else if (fdr < 0 | fdr > 1) { @@ -49,12 +54,21 @@ setMethod( stop("pValue has to be a number between 0 and 1") } if (length(object@kmeans$kpart) == 0) { - stop("run Clustexp before KMClustDiffGenes") + stop("run Clustexp before ClustDiffGenes") } cdiff <- list() x <- object@ndata y <- object@expdata[, names(object@ndata)] - part <- object@kmeans$kpart + if (ran_k) { + part <- object@kmeans$kpart + } else if (ran_m) { + part <- object@MBclusters$clusterid + } else { + stop("Run Clustexp() before running this function") + } + # ====================================================================== + # Operating + # ====================================================================== for (i in 1:max(part)) { if (sum(part == i) == 0) next @@ -224,7 +238,7 @@ setMethod( } return(list(sigDEG, DEGsTable)) } else{ - print(paste0("There are no DEGs with fdr=", fdr)) + print(paste("There are no DEGs with fdr =", fdr)) } } ) \ No newline at end of file diff --git a/R/DIscBIO-generic-MBClustDiffGenes.R b/R/DIscBIO-generic-MBClustDiffGenes.R deleted file mode 100644 index 2ecfc7e..0000000 --- a/R/DIscBIO-generic-MBClustDiffGenes.R +++ /dev/null @@ -1,224 +0,0 @@ -#' @title ClustDiffGenes -#' @rdname MBClustDiffGenes -#' @param object \code{DISCBIO} class object. -#' @param K A numeric value of the number of clusters. -#' @param pValue A numeric value of the p-value. Default is 0.05. -#' @param fdr A numeric value of the false discovery rate. Default is 0.01. -#' @param export A logical vector that allows writing the final gene list in -#' excel file. Default is TRUE. -#' @param quiet if `TRUE`, suppresses intermediate text output -#' @param filename_up Name of the exported "up" file (if `export=TRUE`) -#' @param filename_down Name of the exported "down" file (if `export=TRUE`) -#' @param filename_binom Name of the exported binomial file -#' @param filename_sigdeg Name of the exported sigDEG file -#' @importFrom stats pbinom median -#' @importFrom AnnotationDbi select -#' @import org.Hs.eg.db -#' @export -#' @return A list containing two tables. -setGeneric( - "MBClustDiffGenes", - function( - object, K, pValue = 0.05, fdr = .01, export = FALSE, quiet = FALSE, - filename_up = "Up-DEG-cluster", - filename_down = "Down-DEG-cluster", - filename_binom = "binomial-DEGsTable", - filename_sigdeg = "binomial-sigDEG" - ) { - standardGeneric("MBClustDiffGenes") - } -) -#' @export -#' @rdname MBClustDiffGenes -setMethod( - "MBClustDiffGenes", - signature = "DISCBIO", - definition = function( - object, K, pValue, fdr, export, quiet, filename_up, filename_down, - filename_binom, filename_sigdeg - ) { - if (!is.numeric(fdr)) { - stop("fdr has to be a number between 0 and 1") - } else if (fdr < 0 | fdr > 1) { - stop("fdr has to be a number between 0 and 1") - } - if (!is.numeric(pValue)) { - stop("pValue has to be a number between 0 and 1") - } else if (pValue < 0 | pValue > 1) { - stop("pValue has to be a number between 0 and 1") - } - - cdiff <- list() - x <- object@ndata - y <- object@expdata[, names(object@ndata)] - part <- object@MBclusters$clusterid - for (i in 1:max(part)) { - if (sum(part == i) == 0) - next - m <- apply(x, 1, mean) - n <- - if (sum(part == i) > 1) - apply(x[, part == i], 1, mean) - else - x[, part == i] - no <- - if (sum(part == i) > 1) - median(apply(y[, part == i], 2, sum)) / - median(apply(x[, part == i], 2, sum)) - else - sum(y[, part == i]) / sum(x[, part == i]) - m <- m * no - n <- n * no - pv <- binompval(m / sum(m), sum(n), n) - d <- - data.frame( - mean.all = m, - mean.cl = n, - fc = n / m, - pv = pv - )[order(pv, decreasing = FALSE), ] - cdiff[[i]] <- d[d$pv < pValue, ] - } - DEGsE <- c() - DEGsS <- c() - DEGsTable <- data.frame() - - for (n in 1:K) { - if (length(cdiff[[n]][, 1]) == 0) { - next - } - - if (length(cdiff[[n]][, 1]) > 0) { - p.adj <- p.adjust(cdiff[[n]][, 4], method = "bonferroni") - out <- cbind(cdiff[[n]], p.adj) - out <- subset(out, out[, 5] < fdr) - if (length(out[, 1]) > 0) { - Regulation <- c() - for (i in 1:length(out[, 1])) { - if (out[i, 1] > out[i, 2]) { - Regulation[i] = "Down" - } else{ - Regulation[i] = "Up" - } - } - out <- cbind(out, Regulation) - if (quiet) { - suppressMessages( - geneList <- - AnnotationDbi::select( - org.Hs.eg.db, - keys = keys(org.Hs.eg.db), - columns = c("SYMBOL", "ENSEMBL") - ) - ) - GL <- c(1, "MTRNR2", "ENSG00000210082") - GL1 <- c(1, "MTRNR1", "ENSG00000211459") - geneList <- rbind(geneList, GL, GL1) - } else { - geneList <- - AnnotationDbi::select( - org.Hs.eg.db, - keys = keys(org.Hs.eg.db), - columns = c("SYMBOL", "ENSEMBL") - ) - GL <- c(1, "MTRNR2", "ENSG00000210082") - GL1 <- c(1, "MTRNR1", "ENSG00000211459") - geneList <- rbind(geneList, GL, GL1) - } - genes <- rownames(out) - gene_list <- geneList[, 3] - idx_genes <- is.element(gene_list, genes) - genes2 <- geneList[idx_genes, ] - Final <- cbind(genes, out) - - Final <- - merge( - Final, - genes2, - by.x = "genes", - by.y = "ENSEMBL", - all.x = TRUE - ) - Final <- Final[!duplicated(Final[, 1]),] - Final[is.na(Final[, 9]), c(1, 9)] <- - Final[is.na(Final[, 9]), 1] - rownames(Final) <- Final[, 1] - Final[, 1] <- Final[, 9] - Final <- Final[, -9] - DEGsS <- c(DEGsS, Final[, 1]) - DEGsE <- - c(DEGsE, as.character(rownames(Final))) - Up <- subset(Final, Final[, 7] == "Up") - cols_to_keep <- c( - "Regulation", "genes", "pv", "mean.all", "mean.cl", "fc", "p.adj" - ) - Up <- Up[, cols_to_keep] - Up[, 3] <- rownames(Up) - Up[, 6] <- log2(Up[, 6]) - Up[, 1] <- Up[, 2] - colnames(Up) <- c( - "Genes", "genes", "E.genes", "mean.all", "mean.cl", - "log2.fc", "p.adj" - ) - if (export) { - write.csv( - Up, file = paste0(filename_up, n, ".csv") - ) - } - - Down <- subset(Final, Final[, 7] == "Down") - cols_to_keep <- c( - "Regulation", "genes", "pv", "mean.all", "mean.cl", "fc", "p.adj" - ) - Down <- Down[, cols_to_keep] - Down[, 3] <- rownames(Down) - Down[, 6] <- log2(Down[, 6]) - Down[, 1] <- Down[, 2] - colnames(Down) <- c( - "Genes", - "genes", - "E.genes", - "mean.all", - "mean.cl", - "log2.fc", - "p.adj" - ) - if (export) { - write.csv( - Down, - file = paste0(filename_down, n, ".csv") - ) - } - - sigDEG <- cbind(DEGsE, DEGsS) - if (export) { - write.csv( - sigDEG, file = paste0(filename_sigdeg, ".csv") - ) - } - - DEGsTable[n, 1] <- paste0("Cluster ", n) - DEGsTable[n, 2] <- "Remaining Clusters" - DEGsTable[n, 3] <- length(Up[, 1]) - DEGsTable[n, 4] <- - paste0(filename_up, n, ".csv") - DEGsTable[n, 5] <- length(Down[, 1]) - DEGsTable[n, 6] <- - paste0(filename_down, n, ".csv") - } - } - } - if (length(DEGsTable) > 0) { - colnames(DEGsTable) <- c( - "Target Cluster", "VS", "Gene number", "File name", - "Gene number", "File name" - ) - if (export) { - write.csv(DEGsTable, file = paste0(filename_binom, ".csv")) - } - return(list(sigDEG, DEGsTable)) - } else{ - print(paste0("There are no DEGs with fdr=", fdr)) - } - } -) diff --git a/man/KMClustDiffGenes.Rd b/man/ClustDiffGenes.Rd similarity index 85% rename from man/KMClustDiffGenes.Rd rename to man/ClustDiffGenes.Rd index bd77c73..a98aa4f 100644 --- a/man/KMClustDiffGenes.Rd +++ b/man/ClustDiffGenes.Rd @@ -1,11 +1,11 @@ % Generated by roxygen2: do not edit by hand -% Please edit documentation in R/DIscBIO-generic-KMClustDiffGenes.R -\name{KMClustDiffGenes} -\alias{KMClustDiffGenes} -\alias{KMClustDiffGenes,DISCBIO-method} +% Please edit documentation in R/DIscBIO-generic-ClustDiffGenes.R +\name{ClustDiffGenes} +\alias{ClustDiffGenes} +\alias{ClustDiffGenes,DISCBIO-method} \title{ClustDiffGenes} \usage{ -KMClustDiffGenes( +ClustDiffGenes( object, K, pValue = 0.05, @@ -18,7 +18,7 @@ KMClustDiffGenes( filename_sigdeg = "binomial-sigDEG" ) -\S4method{KMClustDiffGenes}{DISCBIO}( +\S4method{ClustDiffGenes}{DISCBIO}( object, K, pValue = 0.05, diff --git a/man/MBClustDiffGenes.Rd b/man/MBClustDiffGenes.Rd deleted file mode 100644 index c4ebd26..0000000 --- a/man/MBClustDiffGenes.Rd +++ /dev/null @@ -1,61 +0,0 @@ -% Generated by roxygen2: do not edit by hand -% Please edit documentation in R/DIscBIO-generic-MBClustDiffGenes.R -\name{MBClustDiffGenes} -\alias{MBClustDiffGenes} -\alias{MBClustDiffGenes,DISCBIO-method} -\title{ClustDiffGenes} -\usage{ -MBClustDiffGenes( - object, - K, - pValue = 0.05, - fdr = 0.01, - export = FALSE, - quiet = FALSE, - filename_up = "Up-DEG-cluster", - filename_down = "Down-DEG-cluster", - filename_binom = "binomial-DEGsTable", - filename_sigdeg = "binomial-sigDEG" -) - -\S4method{MBClustDiffGenes}{DISCBIO}( - object, - K, - pValue = 0.05, - fdr = 0.01, - export = FALSE, - quiet = FALSE, - filename_up = "Up-DEG-cluster", - filename_down = "Down-DEG-cluster", - filename_binom = "binomial-DEGsTable", - filename_sigdeg = "binomial-sigDEG" -) -} -\arguments{ -\item{object}{\code{DISCBIO} class object.} - -\item{K}{A numeric value of the number of clusters.} - -\item{pValue}{A numeric value of the p-value. Default is 0.05.} - -\item{fdr}{A numeric value of the false discovery rate. Default is 0.01.} - -\item{export}{A logical vector that allows writing the final gene list in -excel file. Default is TRUE.} - -\item{quiet}{if `TRUE`, suppresses intermediate text output} - -\item{filename_up}{Name of the exported "up" file (if `export=TRUE`)} - -\item{filename_down}{Name of the exported "down" file (if `export=TRUE`)} - -\item{filename_binom}{Name of the exported binomial file} - -\item{filename_sigdeg}{Name of the exported sigDEG file} -} -\value{ -A list containing two tables. -} -\description{ -ClustDiffGenes -} diff --git a/tests/testthat/test-DIscBIO.IMP.R b/tests/testthat/test-DIscBIO.IMP.R index e6e838e..f7bff61 100644 --- a/tests/testthat/test-DIscBIO.IMP.R +++ b/tests/testthat/test-DIscBIO.IMP.R @@ -70,7 +70,7 @@ test_that("Cluster plots output is as expexted", { context("Outliers") -Outliers <- FindOutliersKM( +Outliers <- FindOutliers( sc, K=2, outminc=5, outlg=2, probthr=.5*1e-3, thr=2 ^ (-1:-40), outdistquant=.75, plot = FALSE, quiet = TRUE ) From 39c0ab1827bad5eaceef11a5253e9ada0d9201ad Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 3 Aug 2020 15:21:41 +0200 Subject: [PATCH 78/99] Debugging --- NAMESPACE | 1 + R/DIscBIO-generic-ClustDiffGenes.R | 1 + R/DIscBIO-generic-FindOutliers.R | 22 ++++++++++------------ 3 files changed, 12 insertions(+), 12 deletions(-) diff --git a/NAMESPACE b/NAMESPACE index 37a8dc8..65583ab 100644 --- a/NAMESPACE +++ b/NAMESPACE @@ -42,6 +42,7 @@ exportMethods(plotSymbolstSNE) exportMethods(plotexptsneMB) exportMethods(plottSNE) exportMethods(plottsneMB) +import(org.Hs.eg.db) importClassesFrom(SingleCellExperiment,SingleCellExperiment) importFrom(AnnotationDbi,keys) importFrom(NetIndices,GenInd) diff --git a/R/DIscBIO-generic-ClustDiffGenes.R b/R/DIscBIO-generic-ClustDiffGenes.R index a178d08..af757d1 100644 --- a/R/DIscBIO-generic-ClustDiffGenes.R +++ b/R/DIscBIO-generic-ClustDiffGenes.R @@ -12,6 +12,7 @@ #' @param filename_binom Name of the exported binomial file #' @param filename_sigdeg Name of the exported sigDEG file #' @importFrom stats pbinom median +#' @import org.Hs.eg.db #' @rdname ClustDiffGenes #' @return A list containing two tables. #' @export diff --git a/R/DIscBIO-generic-FindOutliers.R b/R/DIscBIO-generic-FindOutliers.R index 131fd13..8c0c51f 100644 --- a/R/DIscBIO-generic-FindOutliers.R +++ b/R/DIscBIO-generic-FindOutliers.R @@ -43,8 +43,8 @@ setMethod( # ====================================================================== # Validating # ====================================================================== - ran_k <- length(object@kmeans$kpart) > 0 - ran_m <- length(object@MBclusters$clusterid) > 0 + ran_k <- length(object@kmeans) > 0 + ran_m <- length(object@MBclusters) > 0 if (ran_k) { clusters <- object@kmeans$kpart } else if (ran_m) { @@ -86,14 +86,13 @@ setMethod( else if (outdistquant < 0 | outdistquant > 1) stop("outdistquant has to be a number between 0 and 1") - object@outlierpar <- - list( - outminc = outminc, - outlg = outlg, - probthr = probthr, - thr = thr, - outdistquant = outdistquant - ) + object@outlierpar <- list( + outminc = outminc, + outlg = outlg, + probthr = probthr, + thr = thr, + outdistquant = outdistquant + ) ### calibrate background model m <- log2(apply(object@fdata, 1, mean)) v <- log2(apply(object@fdata, 1, var)) @@ -243,8 +242,7 @@ setMethod( object@cpart <- cpart object@fcol <- rainbow(max(cpart)) - p <- - clusters[order(clusters, decreasing = FALSE)] + p <- clusters[order(clusters, decreasing = FALSE)] x <- object@out$cprobs[names(p)] fcol <- c("black", "blue", "green", "red", "yellow", "gray") if (plot) { From 859eeb16aa7dd708e5811895deba4a2646f5033d Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 3 Aug 2020 15:32:00 +0200 Subject: [PATCH 79/99] Merged KMclustheatmap and MBclustheatmap --- DESCRIPTION | 2 +- NAMESPACE | 2 +- R/DIscBIO-generic-MBclustheatmap.R | 159 -------------------- R/DIscBIO-generic-clustheatmap.R | 165 +++++++++++++++++++++ man/{MBclustheatmap.Rd => clustheatmap.Rd} | 31 ++-- 5 files changed, 180 insertions(+), 179 deletions(-) delete mode 100644 R/DIscBIO-generic-MBclustheatmap.R create mode 100644 R/DIscBIO-generic-clustheatmap.R rename man/{MBclustheatmap.Rd => clustheatmap.Rd} (59%) diff --git a/DESCRIPTION b/DESCRIPTION index 8b87352..d2d765c 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -66,11 +66,11 @@ Collate: 'DIscBIO-generic-FindOutliers.R' 'DIscBIO-generic-KMclustheatmap.R' 'DIscBIO-generic-KmeanOrder.R' - 'DIscBIO-generic-MBclustheatmap.R' 'DIscBIO-generic-NoiseFiltering.R' 'DIscBIO-generic-Normalizedata.R' 'DIscBIO-generic-PCAplotSymbols.R' 'DIscBIO-generic-PlotmclustMB.R' + 'DIscBIO-generic-clustheatmap.R' 'DIscBIO-generic-comptSNE.R' 'DIscBIO-generic-plotExptSNE.R' 'DIscBIO-generic-plotGap.R' diff --git a/NAMESPACE b/NAMESPACE index 65583ab..a54f5fb 100644 --- a/NAMESPACE +++ b/NAMESPACE @@ -28,11 +28,11 @@ exportMethods(FinalPreprocessing) exportMethods(FindOutliers) exportMethods(KMclustheatmap) exportMethods(KmeanOrder) -exportMethods(MBclustheatmap) exportMethods(NoiseFiltering) exportMethods(Normalizedata) exportMethods(PCAplotSymbols) exportMethods(PlotmclustMB) +exportMethods(clustheatmap) exportMethods(comptSNE) exportMethods(plotExptSNE) exportMethods(plotLabelstSNE) diff --git a/R/DIscBIO-generic-MBclustheatmap.R b/R/DIscBIO-generic-MBclustheatmap.R deleted file mode 100644 index c4b056d..0000000 --- a/R/DIscBIO-generic-MBclustheatmap.R +++ /dev/null @@ -1,159 +0,0 @@ -#' @title Plotting the Model-based clusters in a heatmap representation of the -#' cell-to-cell distances -#' @description This functions plots a heatmap of the distance matrix grouped -#' by clusters. Individual clusters are highlighted with rainbow colors along -#' the x and y-axes. -#' @param object \code{DISCBIO} class object. -#' @param hmethod Agglomeration method used for determining the cluster order -#' from hierarchical clustering of the cluster medoids. This should be one of -#' "ward.D", "ward.D2", "single", "complete", "average". Default is "single". -#' @param plot if `TRUE`, plots the heatmap; otherwise, just prints cclmo -#' @param quiet if `TRUE`, intermediary output is suppressed -#' @param rseed Random integer to fix random results. -#' @importFrom stats hclust as.dist cor kmeans -#' @importFrom cluster clusGap maxSE -#' @return Unless otherwise specified, a heatmap and a vector of the underlying -#' cluster order. -setGeneric( - "MBclustheatmap", - function(object, hmethod="single", plot=TRUE, quiet=FALSE, rseed=NULL) { - standardGeneric("MBclustheatmap") - } -) - -#' @export -#' @rdname MBclustheatmap -setMethod( - "MBclustheatmap", - signature = "DISCBIO", - definition = function(object, hmethod, plot, quiet, rseed) { - x <- object@fdata - object@clusterpar$metric <- "pearson" - y <- clustfun( - object@fdata, - clustnr = 20, - bootnr = 50, - metric = "pearson", - do.gap = TRUE, - SE.method = "Tibs2001SEmax", - SE.factor = .25, - B.gap = 50, - cln = 0, - rseed = rseed, - quiet = quiet - ) - object@distances <- as.matrix(y$di) - part <- object@MBclusters$clusterid - na <- c() - j <- 0 - for (i in 1:max(part)) { - if (sum(part == i) == 0) - next - j <- j + 1 - na <- append(na, i) - d <- x[, part == i] - if (sum(part == i) == 1) - cent <- d - else - cent <- apply(d, 1, mean) - if (j == 1) - tmp <- data.frame(cent) - else - tmp <- cbind(tmp, cent) - } - names(tmp) <- paste("cl", na, sep = ".") - if (max(part) > 1) - cclmo <- - hclust(dist.gen(as.matrix( - dist.gen(t(tmp), method = object@clusterpar$metric) - )), method = hmethod)$order - else - cclmo <- 1 - q <- part - for (i in 1:max(part)) { - q[part == na[cclmo[i]]] <- i - } - part <- q - di <- - as.data.frame(as.matrix(dist.gen(t(object@distances)))) - pto <- part[order(part, decreasing = FALSE)] - ptn <- c() - for (i in 1:max(pto)) { - pt <- - names(pto)[pto == i] - z <- - if (length(pt) == 1) - pt - else - pt[hclust(as.dist(t(di[pt, pt])), method = hmethod)$order] - ptn <- append(ptn, z) - } - col = c("black", "blue", "green", "red", "yellow", "gray") - mi <- min(di, na.rm = TRUE) - ma <- max(di, na.rm = TRUE) - - if (plot) { - layout( - matrix( - data = c(1, 3, 2, 4), - nrow = 2, - ncol = 2 - ), - widths = c(5, 1, 5, 1), - heights = c(5, 1, 1, 1) - ) - ColorRamp <- - colorRampPalette(brewer.pal(n = 7, name = "RdYlBu"))(100) - ColorLevels <- seq(mi, ma, length = length(ColorRamp)) - if (mi == ma) { - ColorLevels <- seq(0.99 * mi, 1.01 * ma, length = length(ColorRamp)) - } - opar <- par(mar = c(3, 5, 2.5, 2)) - on.exit(par(opar)) - image(as.matrix(di[ptn, ptn]), col = ColorRamp, axes = FALSE) - abline(0, 1) - box() - - tmp <- c() - for (u in 1:max(part)) { - ol <- (0:(length(part) - 1) / - (length(part) - 1))[ptn %in% names(x)[part == u]] - points( - rep(0, length(ol)), - ol, - col = col[cclmo[u]], - pch = 15, - cex = .75 - ) - points( - ol, - rep(0, length(ol)), - col = col[cclmo[u]], - pch = 15, - cex = .75 - ) - tmp <- append(tmp, mean(ol)) - } - axis(1, at = tmp, labels = cclmo) - axis(2, at = tmp, labels = cclmo) - opar <- par(mar = c(3, 2.5, 2.5, 2)) - on.exit(par(opar)) - image( - 1, - ColorLevels, - matrix( - data = ColorLevels, - ncol = length(ColorLevels), - nrow = 1 - ), - col = ColorRamp, - xlab = "", - ylab = "", - las = 2, - xaxt = "n" - ) - layout(1) - } - return(cclmo) - } -) \ No newline at end of file diff --git a/R/DIscBIO-generic-clustheatmap.R b/R/DIscBIO-generic-clustheatmap.R new file mode 100644 index 0000000..300be46 --- /dev/null +++ b/R/DIscBIO-generic-clustheatmap.R @@ -0,0 +1,165 @@ +#' @title Plotting clusters in a heatmap representation of the cell distances +#' @description This functions plots a heatmap of the distance matrix grouped +#' by clusters. Individual clusters are highlighted with rainbow colors along +#' the x and y-axes. +#' @param object \code{DISCBIO} class object. +#' @param clustering_method either "k-means" or "model-based" ("k" and "mb" are also accepted) +#' @param hmethod Agglomeration method used for determining the cluster order +#' from hierarchical clustering of the cluster medoids. This should be one of +#' "ward.D", "ward.D2", "single", "complete", "average". Default is "single". +#' @param plot if `TRUE`, plots the heatmap; otherwise, just prints cclmo +#' @return Unless otherwise specified, a heatmap and a vector of the underlying +#' cluster order. +#' @importFrom stats hclust as.dist cor +setGeneric( + "clustheatmap", + function(object, clustering_method = "k-means", hmethod="single", plot=TRUE) + { + standardGeneric("clustheatmap") + } +) + +#' @export +#' @rdname clustheatmap +setMethod( + "clustheatmap", + signature = "DISCBIO", + definition = function(object, clustering_method, hmethod, plot) + { + x <- object@fdata + if (tolower(clustering_method) %in% c("k-means", "k")) { + part <- object@kmeans$kpart + } else if (tolower(clustering_method) %in% c("model-based", "mb")) { + object@clusterpar$metric <- "pearson" + y <- clustfun( + object@fdata, + clustnr = 20, + bootnr = 50, + metric = "pearson", + do.gap = TRUE, + SE.method = "Tibs2001SEmax", + SE.factor = .25, + B.gap = 50, + cln = 0, + rseed = rseed, + quiet = quiet + ) + object@distances <- as.matrix(y$di) + part <- object@MBclusters$clusterid + } + part <- object@kmeans$kpart + na <- c() + j <- 0 + for (i in 1:max(part)) { + if (sum(part == i) == 0) + next + j <- j + 1 + na <- append(na, i) + d <- x[, part == i] + if (sum(part == i) == 1) + cent <- d + else + cent <- apply(d, 1, mean) + if (j == 1) + tmp <- data.frame(cent) + else + tmp <- cbind(tmp, cent) + } + names(tmp) <- paste("cl", na, sep = ".") + if (max(part) > 1) + cclmo <- + hclust(dist.gen(as.matrix( + dist.gen(t(tmp), method = object@clusterpar$metric) + )), method = hmethod)$order + else + cclmo <- 1 + q <- part + for (i in 1:max(part)) { + q[part == na[cclmo[i]]] <- i + } + part <- q + di <- as.data.frame(as.matrix(dist.gen(t(object@distances)))) + pto <- part[order(part, decreasing = FALSE)] + ptn <- c() + for (i in 1:max(pto)) { + pt <- + names(pto)[pto == i] + z <- + if (length(pt) == 1) + pt + else + pt[hclust(as.dist(t(di[pt, pt])), method = hmethod)$order] + ptn <- append(ptn, z) + } + col = c("black", "blue", "green", "red", "yellow", "gray") + mi <- min(di, na.rm = TRUE) + ma <- max(di, na.rm = TRUE) + if (plot) { + layout( + matrix( + data = c(1, 3, 2, 4), + nrow = 2, + ncol = 2 + ), + widths = c(5, 1, 5, 1), + heights = c(5, 1, 1, 1) + ) + ColorRamp <- colorRampPalette( + brewer.pal(n = 7, name = "RdYlBu") + )(100) + ColorLevels <- seq(mi, ma, length = length(ColorRamp)) + if (mi == ma) { + ColorLevels <- seq( + 0.99 * mi, 1.01 * ma, length = length(ColorRamp) + ) + } + + opar <- par(mar = c(3, 5, 2.5, 2)) + on.exit(par(opar)) + image(as.matrix(di[ptn, ptn]), col = ColorRamp, axes = FALSE) + abline(0, 1) + box() + + tmp <- c() + for (u in 1:max(part)) { + ol <- (0:(length(part) - 1) / + (length(part) - 1))[ptn %in% names(x)[part == u]] + points( + rep(0, length(ol)), + ol, + col = col[cclmo[u]], + pch = 15, + cex = .75 + ) + points( + ol, + rep(0, length(ol)), + col = col[cclmo[u]], + pch = 15, + cex = .75 + ) + tmp <- append(tmp, mean(ol)) + } + axis(1, at = tmp, labels = cclmo) + axis(2, at = tmp, labels = cclmo) + opar <- par(mar = c(3, 2.5, 2.5, 2)) + on.exit(par(opar)) + image( + 1, + ColorLevels, + matrix( + data = ColorLevels, + ncol = length(ColorLevels), + nrow = 1 + ), + col = ColorRamp, + xlab = "", + ylab = "", + las = 2, + xaxt = "n" + ) + layout(1) + } + return(cclmo) + } +) \ No newline at end of file diff --git a/man/MBclustheatmap.Rd b/man/clustheatmap.Rd similarity index 59% rename from man/MBclustheatmap.Rd rename to man/clustheatmap.Rd index 9d993c4..9b8e1e5 100644 --- a/man/MBclustheatmap.Rd +++ b/man/clustheatmap.Rd @@ -1,39 +1,34 @@ % Generated by roxygen2: do not edit by hand -% Please edit documentation in R/DIscBIO-generic-MBclustheatmap.R -\name{MBclustheatmap} -\alias{MBclustheatmap} -\alias{MBclustheatmap,DISCBIO-method} -\title{Plotting the Model-based clusters in a heatmap representation of the - cell-to-cell distances} +% Please edit documentation in R/DIscBIO-generic-clustheatmap.R +\name{clustheatmap} +\alias{clustheatmap} +\alias{clustheatmap,DISCBIO-method} +\title{Plotting clusters in a heatmap representation of the cell distances} \usage{ -MBclustheatmap( +clustheatmap( object, + clustering_method = "k-means", hmethod = "single", - plot = TRUE, - quiet = FALSE, - rseed = NULL + plot = TRUE ) -\S4method{MBclustheatmap}{DISCBIO}( +\S4method{clustheatmap}{DISCBIO}( object, + clustering_method = "k-means", hmethod = "single", - plot = TRUE, - quiet = FALSE, - rseed = NULL + plot = TRUE ) } \arguments{ \item{object}{\code{DISCBIO} class object.} +\item{clustering_method}{either "k-means" or "model-based" ("k" and "mb" are also accepted)} + \item{hmethod}{Agglomeration method used for determining the cluster order from hierarchical clustering of the cluster medoids. This should be one of "ward.D", "ward.D2", "single", "complete", "average". Default is "single".} \item{plot}{if `TRUE`, plots the heatmap; otherwise, just prints cclmo} - -\item{quiet}{if `TRUE`, intermediary output is suppressed} - -\item{rseed}{Random integer to fix random results.} } \value{ Unless otherwise specified, a heatmap and a vector of the underlying From 4e26aadc0d93e1b3a8e8cba2502e771b267f2aa6 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 3 Aug 2020 15:42:14 +0200 Subject: [PATCH 80/99] Debugged clustheatmap --- DESCRIPTION | 1 - NAMESPACE | 1 - R/DIscBIO-generic-KMclustheatmap.R | 146 ----------------------------- R/DIscBIO-generic-clustheatmap.R | 10 +- man/KMclustheatmap.Rd | 30 ------ man/clustheatmap.Rd | 13 +-- 6 files changed, 16 insertions(+), 185 deletions(-) delete mode 100644 R/DIscBIO-generic-KMclustheatmap.R delete mode 100644 man/KMclustheatmap.Rd diff --git a/DESCRIPTION b/DESCRIPTION index d2d765c..abd3ce7 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -64,7 +64,6 @@ Collate: 'DIscBIO-generic-Exprmclust.R' 'DIscBIO-generic-FinalPreprocessing.R' 'DIscBIO-generic-FindOutliers.R' - 'DIscBIO-generic-KMclustheatmap.R' 'DIscBIO-generic-KmeanOrder.R' 'DIscBIO-generic-NoiseFiltering.R' 'DIscBIO-generic-Normalizedata.R' diff --git a/NAMESPACE b/NAMESPACE index a54f5fb..2004d49 100644 --- a/NAMESPACE +++ b/NAMESPACE @@ -26,7 +26,6 @@ exportMethods(DEGanalysis2clust) exportMethods(Exprmclust) exportMethods(FinalPreprocessing) exportMethods(FindOutliers) -exportMethods(KMclustheatmap) exportMethods(KmeanOrder) exportMethods(NoiseFiltering) exportMethods(Normalizedata) diff --git a/R/DIscBIO-generic-KMclustheatmap.R b/R/DIscBIO-generic-KMclustheatmap.R deleted file mode 100644 index 350c1b3..0000000 --- a/R/DIscBIO-generic-KMclustheatmap.R +++ /dev/null @@ -1,146 +0,0 @@ -#' @title Plotting the K-means clusters in a heatmap representation of the -#' cell-to-cell distances -#' @description This functions plots a heatmap of the distance matrix grouped -#' by clusters. Individual clusters are highlighted with rainbow colors along -#' the x and y-axes. -#' @param object \code{DISCBIO} class object. -#' @param hmethod Agglomeration method used for determining the cluster order -#' from hierarchical clustering of the cluster medoids. This should be one of -#' "ward.D", "ward.D2", "single", "complete", "average". Default is "single". -#' @param plot if `TRUE`, plots the heatmap; otherwise, just prints cclmo -#' @return Unless otherwise specified, a heatmap and a vector of the underlying -#' cluster order. -#' @importFrom stats hclust as.dist cor -setGeneric( - "KMclustheatmap", - function( - object, hmethod = "single", plot = TRUE - ) - standardGeneric("KMclustheatmap") -) - -#' @export -#' @rdname KMclustheatmap -setMethod( - "KMclustheatmap", - signature = "DISCBIO", - definition = function(object, hmethod, plot = TRUE) { - x <- object@fdata - part <- object@kmeans$kpart - na <- c() - j <- 0 - for (i in 1:max(part)) { - if (sum(part == i) == 0) - next - j <- j + 1 - na <- append(na, i) - d <- x[, part == i] - if (sum(part == i) == 1) - cent <- d - else - cent <- apply(d, 1, mean) - if (j == 1) - tmp <- data.frame(cent) - else - tmp <- cbind(tmp, cent) - } - names(tmp) <- paste("cl", na, sep = ".") - if (max(part) > 1) - cclmo <- - hclust(dist.gen(as.matrix( - dist.gen(t(tmp), method = object@clusterpar$metric) - )), method = hmethod)$order - else - cclmo <- 1 - q <- part - for (i in 1:max(part)) { - q[part == na[cclmo[i]]] <- i - } - part <- q - di <- - as.data.frame(as.matrix(dist.gen(t( - object@distances - )))) - pto <- part[order(part, decreasing = FALSE)] - ptn <- c() - for (i in 1:max(pto)) { - pt <- - names(pto)[pto == i] - z <- - if (length(pt) == 1) - pt - else - pt[hclust(as.dist(t(di[pt, pt])), method = hmethod)$order] - ptn <- append(ptn, z) - } - col = c("black", "blue", "green", "red", "yellow", "gray") - mi <- min(di, na.rm = TRUE) - ma <- max(di, na.rm = TRUE) - if (plot) { - layout( - matrix( - data = c(1, 3, 2, 4), - nrow = 2, - ncol = 2 - ), - widths = c(5, 1, 5, 1), - heights = c(5, 1, 1, 1) - ) - ColorRamp <- - colorRampPalette(brewer.pal(n = 7, name = "RdYlBu"))(100) - ColorLevels <- seq(mi, ma, length = length(ColorRamp)) - if (mi == ma) { - ColorLevels <- seq( - 0.99 * mi, 1.01 * ma, length = length(ColorRamp) - ) - } - - opar <- par(mar = c(3, 5, 2.5, 2)) - on.exit(par(opar)) - image(as.matrix(di[ptn, ptn]), col = ColorRamp, axes = FALSE) - abline(0, 1) - box() - - tmp <- c() - for (u in 1:max(part)) { - ol <- (0:(length(part) - 1) / - (length(part) - 1))[ptn %in% names(x)[part == u]] - points( - rep(0, length(ol)), - ol, - col = col[cclmo[u]], - pch = 15, - cex = .75 - ) - points( - ol, - rep(0, length(ol)), - col = col[cclmo[u]], - pch = 15, - cex = .75 - ) - tmp <- append(tmp, mean(ol)) - } - axis(1, at = tmp, labels = cclmo) - axis(2, at = tmp, labels = cclmo) - opar <- par(mar = c(3, 2.5, 2.5, 2)) - on.exit(par(opar)) - image( - 1, - ColorLevels, - matrix( - data = ColorLevels, - ncol = length(ColorLevels), - nrow = 1 - ), - col = ColorRamp, - xlab = "", - ylab = "", - las = 2, - xaxt = "n" - ) - layout(1) - } - return(cclmo) - } -) \ No newline at end of file diff --git a/R/DIscBIO-generic-clustheatmap.R b/R/DIscBIO-generic-clustheatmap.R index 300be46..d71538e 100644 --- a/R/DIscBIO-generic-clustheatmap.R +++ b/R/DIscBIO-generic-clustheatmap.R @@ -7,13 +7,21 @@ #' @param hmethod Agglomeration method used for determining the cluster order #' from hierarchical clustering of the cluster medoids. This should be one of #' "ward.D", "ward.D2", "single", "complete", "average". Default is "single". +#' @param quiet if `TRUE`, intermediary output is suppressed +#' @param rseed Random integer to fix random results. #' @param plot if `TRUE`, plots the heatmap; otherwise, just prints cclmo #' @return Unless otherwise specified, a heatmap and a vector of the underlying #' cluster order. #' @importFrom stats hclust as.dist cor setGeneric( "clustheatmap", - function(object, clustering_method = "k-means", hmethod="single", plot=TRUE) + function( + object, + clustering_method = "k-means", + hmethod="single", + rseed=NULL, + quiet=FALSE, + plot=TRUE) { standardGeneric("clustheatmap") } diff --git a/man/KMclustheatmap.Rd b/man/KMclustheatmap.Rd deleted file mode 100644 index 9a1c4c8..0000000 --- a/man/KMclustheatmap.Rd +++ /dev/null @@ -1,30 +0,0 @@ -% Generated by roxygen2: do not edit by hand -% Please edit documentation in R/DIscBIO-generic-KMclustheatmap.R -\name{KMclustheatmap} -\alias{KMclustheatmap} -\alias{KMclustheatmap,DISCBIO-method} -\title{Plotting the K-means clusters in a heatmap representation of the - cell-to-cell distances} -\usage{ -KMclustheatmap(object, hmethod = "single", plot = TRUE) - -\S4method{KMclustheatmap}{DISCBIO}(object, hmethod = "single", plot = TRUE) -} -\arguments{ -\item{object}{\code{DISCBIO} class object.} - -\item{hmethod}{Agglomeration method used for determining the cluster order -from hierarchical clustering of the cluster medoids. This should be one of -"ward.D", "ward.D2", "single", "complete", "average". Default is "single".} - -\item{plot}{if `TRUE`, plots the heatmap; otherwise, just prints cclmo} -} -\value{ -Unless otherwise specified, a heatmap and a vector of the underlying - cluster order. -} -\description{ -This functions plots a heatmap of the distance matrix grouped - by clusters. Individual clusters are highlighted with rainbow colors along - the x and y-axes. -} diff --git a/man/clustheatmap.Rd b/man/clustheatmap.Rd index 9b8e1e5..9088413 100644 --- a/man/clustheatmap.Rd +++ b/man/clustheatmap.Rd @@ -9,15 +9,12 @@ clustheatmap( object, clustering_method = "k-means", hmethod = "single", + rseed = NULL, + quiet = FALSE, plot = TRUE ) -\S4method{clustheatmap}{DISCBIO}( - object, - clustering_method = "k-means", - hmethod = "single", - plot = TRUE -) +\S4method{clustheatmap}{DISCBIO}(object, clustering_method, hmethod, plot) } \arguments{ \item{object}{\code{DISCBIO} class object.} @@ -28,6 +25,10 @@ clustheatmap( from hierarchical clustering of the cluster medoids. This should be one of "ward.D", "ward.D2", "single", "complete", "average". Default is "single".} +\item{rseed}{Random integer to fix random results.} + +\item{quiet}{if `TRUE`, intermediary output is suppressed} + \item{plot}{if `TRUE`, plots the heatmap; otherwise, just prints cclmo} } \value{ From 3f666d287001cf2ed5d5c559b97e7396aea5373e Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 3 Aug 2020 15:42:25 +0200 Subject: [PATCH 81/99] Merged plotexptsneMB into plotExptSNE --- DESCRIPTION | 1 - NAMESPACE | 1 - R/DIscBIO-generic-plotExptSNE.R | 29 ++++++++--- R/DIscBIO-generic-plotexptsneMB.R | 87 ------------------------------- man/plotExptSNE.Rd | 2 +- man/plotexptsneMB.Rd | 30 ----------- 6 files changed, 22 insertions(+), 128 deletions(-) delete mode 100644 R/DIscBIO-generic-plotexptsneMB.R delete mode 100644 man/plotexptsneMB.Rd diff --git a/DESCRIPTION b/DESCRIPTION index abd3ce7..f2a5879 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -77,7 +77,6 @@ Collate: 'DIscBIO-generic-plotOrderTsne.R' 'DIscBIO-generic-plotSilhouette.R' 'DIscBIO-generic-plotSymbolstSNE.R' - 'DIscBIO-generic-plotexptsneMB.R' 'DIscBIO-generic-plottSNE.R' 'DIscBIO-generic-plottsneMB.R' 'J48DT.R' diff --git a/NAMESPACE b/NAMESPACE index 2004d49..826c108 100644 --- a/NAMESPACE +++ b/NAMESPACE @@ -38,7 +38,6 @@ exportMethods(plotLabelstSNE) exportMethods(plotOrderTsne) exportMethods(plotSilhouette) exportMethods(plotSymbolstSNE) -exportMethods(plotexptsneMB) exportMethods(plottSNE) exportMethods(plottsneMB) import(org.Hs.eg.db) diff --git a/R/DIscBIO-generic-plotExptSNE.R b/R/DIscBIO-generic-plotExptSNE.R index b67e418..0fed04b 100644 --- a/R/DIscBIO-generic-plotExptSNE.R +++ b/R/DIscBIO-generic-plotExptSNE.R @@ -1,4 +1,4 @@ -#' @title Highlighting gene expression in K-means clustering in the t-SNE map +#' @title Highlighting gene expression in the t-SNE map #' @description The t-SNE map representation can also be used to analyze #' expression of a gene or a group of genes, to investigate cluster specific #' gene expression patterns @@ -20,20 +20,33 @@ setMethod( "plotExptSNE", signature = "DISCBIO", definition = function(object, g, n = NULL) { - if (length(object@tsne) == 0) + # ====================================================================== + # Validation + # ====================================================================== + ran_k <- length(object@tsne) > 0 + ran_m <- length(object@MBtsne) > 0 + if (ran_k) { + x <- object@tsne + } else if (ran_m) { + x <- object@MBtsne + } else { stop("run comptSNE before plotExptSNE") + } if (length(intersect(g, rownames(object@ndata))) < length(unique(g))) stop( "second argument does not correspond to set of rownames slot", "ndata of SCseq object" ) - if (is.null(n)) - n <- g[1] + if (is.null(n)) n <- g[1] + # ====================================================================== + # Plotting + # ====================================================================== l <- apply(object@ndata[g, ] - .1, 2, sum) + .1 mi <- min(l, na.rm = TRUE) ma <- max(l, na.rm = TRUE) - ColorRamp <- - colorRampPalette(rev(brewer.pal(n = 7, name = "RdYlBu")))(100) + ColorRamp <- colorRampPalette( + rev(brewer.pal(n = 7, name = "RdYlBu")) + )(100) ColorLevels <- seq(mi, ma, length = length(ColorRamp)) v <- round((l - mi) / (ma - mi) * 99 + 1, 0) layout( @@ -59,8 +72,8 @@ setMethod( ) for (k in 1:length(v)) { points( - object@tsne[k, 1], - object@tsne[k, 2], + x[k, 1], + x[k, 2], col = ColorRamp[v[k]], pch = 20, cex = 1.5 diff --git a/R/DIscBIO-generic-plotexptsneMB.R b/R/DIscBIO-generic-plotexptsneMB.R deleted file mode 100644 index dcb9b38..0000000 --- a/R/DIscBIO-generic-plotexptsneMB.R +++ /dev/null @@ -1,87 +0,0 @@ -#' @title Highlighting gene expression in Model-based clustering in the t-SNE -#' map -#' @description The t-SNE map representation can also be used to analyze -#' expression of a gene or a group of genes, to investigate cluster specific -#' gene expression patterns -#' @param object \code{DISCBIO} class object. -#' @param g Individual gene name or vector with a group of gene names -#' corresponding to a subset of valid row names of the \code{ndata} slot of -#' the \code{DISCBIO} object. -#' @param n String of characters representing the title of the plot. Default is -#' NULL and the first element of \code{g} is chosen. -#' @return t-SNE plot for one particular gene -setGeneric("plotexptsneMB", function(object, g, n = NULL) - standardGeneric("plotexptsneMB")) - -#' @export -#' @rdname plotexptsneMB -setMethod( - "plotexptsneMB", - signature = "DISCBIO", - definition = function(object, g, n = NULL) { - if (length(object@MBtsne) == 0) - stop("run comptsne before plotexptsneMB") - if (length(intersect(g, rownames(object@ndata))) < length(unique(g))) - stop( - "second argument does not correspond to set of rownames", - "slot ndata of SCseq object" - ) - if (is.null(n)) - n <- g[1] - l <- apply(object@ndata[g, ] - .1, 2, sum) + .1 - - mi <- min(l, na.rm = TRUE) - ma <- max(l, na.rm = TRUE) - ColorRamp <- - colorRampPalette(rev(brewer.pal(n = 7, name = "RdYlBu")))(100) - ColorLevels <- seq(mi, ma, length = length(ColorRamp)) - v <- round((l - mi) / (ma - mi) * 99 + 1, 0) - layout( - matrix( - data = c(1, 3, 2, 4), - nrow = 2, - ncol = 2 - ), - widths = c(5, 1, 5, 1), - heights = c(5, 1, 1, 1) - ) - opar <- par(mar = c(3, 5, 2.5, 2)) - on.exit(par(opar)) - plot( - object@MBtsne, - xlab = "Dim 1", - ylab = "Dim 2", - main = n, - pch = 20, - cex = 0, - col = "grey", - las = 1 - ) - for (k in 1:length(v)) { - points( - object@MBtsne[k, 1], - object@MBtsne[k, 2], - col = ColorRamp[v[k]], - pch = 20, - cex = 1.5 - ) - } - opar <- par(mar = c(3, 2.5, 2.5, 2)) - on.exit(par(opar)) - image( - 1, - ColorLevels, - matrix( - data = ColorLevels, - ncol = length(ColorLevels), - nrow = 1 - ), - col = ColorRamp, - xlab = "", - ylab = "", - las = 1, - xaxt = "n" - ) - layout(1) - } -) \ No newline at end of file diff --git a/man/plotExptSNE.Rd b/man/plotExptSNE.Rd index 56d250b..29fe5b7 100644 --- a/man/plotExptSNE.Rd +++ b/man/plotExptSNE.Rd @@ -3,7 +3,7 @@ \name{plotExptSNE} \alias{plotExptSNE} \alias{plotExptSNE,DISCBIO-method} -\title{Highlighting gene expression in K-means clustering in the t-SNE map} +\title{Highlighting gene expression in the t-SNE map} \usage{ plotExptSNE(object, g, n = NULL) diff --git a/man/plotexptsneMB.Rd b/man/plotexptsneMB.Rd deleted file mode 100644 index e7eb5c9..0000000 --- a/man/plotexptsneMB.Rd +++ /dev/null @@ -1,30 +0,0 @@ -% Generated by roxygen2: do not edit by hand -% Please edit documentation in R/DIscBIO-generic-plotexptsneMB.R -\name{plotexptsneMB} -\alias{plotexptsneMB} -\alias{plotexptsneMB,DISCBIO-method} -\title{Highlighting gene expression in Model-based clustering in the t-SNE - map} -\usage{ -plotexptsneMB(object, g, n = NULL) - -\S4method{plotexptsneMB}{DISCBIO}(object, g, n = NULL) -} -\arguments{ -\item{object}{\code{DISCBIO} class object.} - -\item{g}{Individual gene name or vector with a group of gene names -corresponding to a subset of valid row names of the \code{ndata} slot of -the \code{DISCBIO} object.} - -\item{n}{String of characters representing the title of the plot. Default is -NULL and the first element of \code{g} is chosen.} -} -\value{ -t-SNE plot for one particular gene -} -\description{ -The t-SNE map representation can also be used to analyze - expression of a gene or a group of genes, to investigate cluster specific - gene expression patterns -} From 4d184fb1eedc91a79d00d542d476799a3229b2ba Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 3 Aug 2020 15:51:48 +0200 Subject: [PATCH 82/99] Merged MmeanOrder and MB_Order into pseudoTimeOrdering --- DESCRIPTION | 4 +- NAMESPACE | 2 +- ...er.R => DIscBIO-generic-clusteringOrder.R} | 0 R/DIscBIO-generic-pseudoTimeOrdering.R | 56 +++++++++++++++++++ R/MB_Order.R | 34 ----------- man/KmeanOrder.Rd | 2 +- man/MB_Order.Rd | 30 ---------- man/pseudoTimeOrdering.Rd | 38 +++++++++++++ 8 files changed, 98 insertions(+), 68 deletions(-) rename R/{DIscBIO-generic-KmeanOrder.R => DIscBIO-generic-clusteringOrder.R} (100%) create mode 100644 R/DIscBIO-generic-pseudoTimeOrdering.R delete mode 100644 R/MB_Order.R delete mode 100644 man/MB_Order.Rd create mode 100644 man/pseudoTimeOrdering.Rd diff --git a/DESCRIPTION b/DESCRIPTION index f2a5879..93769f2 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -64,11 +64,11 @@ Collate: 'DIscBIO-generic-Exprmclust.R' 'DIscBIO-generic-FinalPreprocessing.R' 'DIscBIO-generic-FindOutliers.R' - 'DIscBIO-generic-KmeanOrder.R' 'DIscBIO-generic-NoiseFiltering.R' 'DIscBIO-generic-Normalizedata.R' 'DIscBIO-generic-PCAplotSymbols.R' 'DIscBIO-generic-PlotmclustMB.R' + 'DIscBIO-generic-clusteringOrder.R' 'DIscBIO-generic-clustheatmap.R' 'DIscBIO-generic-comptSNE.R' 'DIscBIO-generic-plotExptSNE.R' @@ -79,10 +79,10 @@ Collate: 'DIscBIO-generic-plotSymbolstSNE.R' 'DIscBIO-generic-plottSNE.R' 'DIscBIO-generic-plottsneMB.R' + 'DIscBIO-generic-pseudoTimeOrdering.R' 'J48DT.R' 'J48DTeval.R' 'Jaccard.R' - 'MB_Order.R' 'NetAnalysis.R' 'Networking.R' 'PPI.R' diff --git a/NAMESPACE b/NAMESPACE index 826c108..89ad442 100644 --- a/NAMESPACE +++ b/NAMESPACE @@ -6,7 +6,6 @@ export(DISCBIO2SingleCellExperiment) export(J48DT) export(J48DTeval) export(Jaccard) -export(MB_Order) export(NetAnalysis) export(Networking) export(PPI) @@ -40,6 +39,7 @@ exportMethods(plotSilhouette) exportMethods(plotSymbolstSNE) exportMethods(plottSNE) exportMethods(plottsneMB) +exportMethods(pseudoTimeOrdering) import(org.Hs.eg.db) importClassesFrom(SingleCellExperiment,SingleCellExperiment) importFrom(AnnotationDbi,keys) diff --git a/R/DIscBIO-generic-KmeanOrder.R b/R/DIscBIO-generic-clusteringOrder.R similarity index 100% rename from R/DIscBIO-generic-KmeanOrder.R rename to R/DIscBIO-generic-clusteringOrder.R diff --git a/R/DIscBIO-generic-pseudoTimeOrdering.R b/R/DIscBIO-generic-pseudoTimeOrdering.R new file mode 100644 index 0000000..2be44e3 --- /dev/null +++ b/R/DIscBIO-generic-pseudoTimeOrdering.R @@ -0,0 +1,56 @@ +#' @title Pseudo-time ordering +#' @description This function takes the exact output of exprmclust function and +#' construct Pseudo-time ordering by mapping all cells onto the path that +#' connects cluster centers. +#' @param object \code{DISCBIO} class object. +#' @param quiet if `TRUE`, suppresses intermediary output +#' @param export if `TRUE`, exports order table to csv +#' @param filename Name of the exported file (if `export=TRUE`) +#' @importFrom TSCAN TSCANorder +#' @return The DISCBIO-class object input with the kordering slot filled. +setGeneric("pseudoTimeOrdering", function( + object, + quiet=FALSE, + export=FALSE, + filename="Cellular_pseudo-time_ordering" + ) + { + standardGeneric("pseudoTimeOrdering") + } +) + +#' @export +#' @rdname pseudoTimeOrdering +setMethod( + "pseudoTimeOrdering", + signature = "DISCBIO", + definition = function(object, quiet, export, filename) { + # ====================================================================== + # Validating + # ====================================================================== + ran_k <- length(object@kmeans$kpart) > 0 + ran_m <- length(object@MBclusters) > 0 + if (ran_k) { + Names <- object@cpart + lpsmclust <- Exprmclust(Obj, K = 4, reduce = F, cluster = Clusters) + lpsorder <- TSCANorder(lpsmclust) + } else if (ran_m) { + Names <- names(object@MBclusters$clusterid) + lpsmclust <- object@MBclusters + lpsorder <- TSCANorder(lpsmclust) + } else { + stop("run clustexp before this pseudoTimeOrdering") + } + sampleNames <- colnames(object@fdata) + orderID <- lpsorder + order <- c(1:length(lpsorder)) + orderTable <- data.frame(order, orderID) + if (export) write.csv(orderTable, file = paste0(filename, ".csv")) + if (!quiet) print(orderTable) + FinalOrder <- orderTable[match(sampleNames, orderTable$orderID), ] + out_order <- FinalOrder[, 1] + names(out_order) <- names(Names) + object@kordering <- out_order + return(object) + } +) \ No newline at end of file diff --git a/R/MB_Order.R b/R/MB_Order.R deleted file mode 100644 index c8de73e..0000000 --- a/R/MB_Order.R +++ /dev/null @@ -1,34 +0,0 @@ -#' @title Pseudo-time ordering based on Model-based clusters -#' @description This function takes the exact output of exprmclust function and -#' construct Pseudo-time ordering by mapping all cells onto the path that -#' connects cluster centers. -#' @export -#' @param object \code{DISCBIO} class object. -#' @param quiet if `TRUE`, intermediary output is suppressed -#' @param export if `TRUE`, exports the results as a CSV file -#' @param filename Name of the exported file (if `export=TRUE`) -#' @importFrom TSCAN TSCANorder -#' @return The DISCBIO-class object input with the MBordering slot filled. -MB_Order <- function( - object, quiet = FALSE, export = FALSE, - filename = "Cellular_pseudo-time_ordering_based_on_Model-based_clusters" -) { - data = object@MBclusters - lpsorderMB <- TSCANorder(data) - Names <- names(object@MBclusters$clusterid) - sampleNames <- colnames(object@fdata) - orderID <- lpsorderMB - order <- c(1:length(lpsorderMB)) - orderTableMB <- data.frame(order, orderID) - if (export) { - write.csv(orderTableMB, file = paste0(filename, ".csv")) - } - if (!quiet) { - print(orderTableMB) - } - FinalOrder <- orderTableMB[match(sampleNames, orderTableMB$orderID), ] - MBordering <- FinalOrder[, 1] - names(MBordering) <- names(Names) - object@MBordering <- MBordering - return(object) -} \ No newline at end of file diff --git a/man/KmeanOrder.Rd b/man/KmeanOrder.Rd index 4c97fbe..165889c 100644 --- a/man/KmeanOrder.Rd +++ b/man/KmeanOrder.Rd @@ -1,5 +1,5 @@ % Generated by roxygen2: do not edit by hand -% Please edit documentation in R/DIscBIO-generic-KmeanOrder.R +% Please edit documentation in R/DIscBIO-generic-clusteringOrder.R \name{KmeanOrder} \alias{KmeanOrder} \alias{KmeanOrder,DISCBIO-method} diff --git a/man/MB_Order.Rd b/man/MB_Order.Rd deleted file mode 100644 index 70c2739..0000000 --- a/man/MB_Order.Rd +++ /dev/null @@ -1,30 +0,0 @@ -% Generated by roxygen2: do not edit by hand -% Please edit documentation in R/MB_Order.R -\name{MB_Order} -\alias{MB_Order} -\title{Pseudo-time ordering based on Model-based clusters} -\usage{ -MB_Order( - object, - quiet = FALSE, - export = FALSE, - filename = "Cellular_pseudo-time_ordering_based_on_Model-based_clusters" -) -} -\arguments{ -\item{object}{\code{DISCBIO} class object.} - -\item{quiet}{if `TRUE`, intermediary output is suppressed} - -\item{export}{if `TRUE`, exports the results as a CSV file} - -\item{filename}{Name of the exported file (if `export=TRUE`)} -} -\value{ -The DISCBIO-class object input with the MBordering slot filled. -} -\description{ -This function takes the exact output of exprmclust function and - construct Pseudo-time ordering by mapping all cells onto the path that - connects cluster centers. -} diff --git a/man/pseudoTimeOrdering.Rd b/man/pseudoTimeOrdering.Rd new file mode 100644 index 0000000..8a57157 --- /dev/null +++ b/man/pseudoTimeOrdering.Rd @@ -0,0 +1,38 @@ +% Generated by roxygen2: do not edit by hand +% Please edit documentation in R/DIscBIO-generic-pseudoTimeOrdering.R +\name{pseudoTimeOrdering} +\alias{pseudoTimeOrdering} +\alias{pseudoTimeOrdering,DISCBIO-method} +\title{Pseudo-time ordering} +\usage{ +pseudoTimeOrdering( + object, + quiet = FALSE, + export = FALSE, + filename = "Cellular_pseudo-time_ordering" +) + +\S4method{pseudoTimeOrdering}{DISCBIO}( + object, + quiet = FALSE, + export = FALSE, + filename = "Cellular_pseudo-time_ordering" +) +} +\arguments{ +\item{object}{\code{DISCBIO} class object.} + +\item{quiet}{if `TRUE`, suppresses intermediary output} + +\item{export}{if `TRUE`, exports order table to csv} + +\item{filename}{Name of the exported file (if `export=TRUE`)} +} +\value{ +The DISCBIO-class object input with the kordering slot filled. +} +\description{ +This function takes the exact output of exprmclust function and + construct Pseudo-time ordering by mapping all cells onto the path that + connects cluster centers. +} From f06f09f3974728744870eb9ac4bb9f0feee78d16 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 3 Aug 2020 15:51:56 +0200 Subject: [PATCH 83/99] Unit test debugging --- tests/testthat/test-DIscBIO.IMP.R | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/tests/testthat/test-DIscBIO.IMP.R b/tests/testthat/test-DIscBIO.IMP.R index f7bff61..1a7d219 100644 --- a/tests/testthat/test-DIscBIO.IMP.R +++ b/tests/testthat/test-DIscBIO.IMP.R @@ -61,7 +61,7 @@ test_that("Cluster plots output is as expexted", { tolerance = .01 ) expect_equal( - object = KMclustheatmap(sc, hmethod = "single", plot = FALSE), + object = clustheatmap(sc, hmethod = "single", plot = FALSE), expected = c(1, 2) ) }) From de296b6c2be1308ff138b167be1cf0fa4db66385 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 3 Aug 2020 16:11:07 +0200 Subject: [PATCH 84/99] Debugging --- man/clustheatmap.Rd | 9 ++++++++- man/plotSilhouette.Rd | 6 +----- 2 files changed, 9 insertions(+), 6 deletions(-) diff --git a/man/clustheatmap.Rd b/man/clustheatmap.Rd index 9088413..9dee417 100644 --- a/man/clustheatmap.Rd +++ b/man/clustheatmap.Rd @@ -14,7 +14,14 @@ clustheatmap( plot = TRUE ) -\S4method{clustheatmap}{DISCBIO}(object, clustering_method, hmethod, plot) +\S4method{clustheatmap}{DISCBIO}( + object, + clustering_method = "k-means", + hmethod = "single", + rseed = NULL, + quiet = FALSE, + plot = TRUE +) } \arguments{ \item{object}{\code{DISCBIO} class object.} diff --git a/man/plotSilhouette.Rd b/man/plotSilhouette.Rd index 074dcb4..7e99e21 100644 --- a/man/plotSilhouette.Rd +++ b/man/plotSilhouette.Rd @@ -1,17 +1,13 @@ % Generated by roxygen2: do not edit by hand -% Please edit documentation in R/DIscBIO-generic-plotSilhouette.R, -% R/DIscBIO-generic-plottSNE.R +% Please edit documentation in R/DIscBIO-generic-plotSilhouette.R \name{plotSilhouette} \alias{plotSilhouette} \alias{plotSilhouette,DISCBIO-method} -\alias{plottSNE,DISCBIO-method} \title{Silhouette Plot for K-means clustering} \usage{ plotSilhouette(object, K) \S4method{plotSilhouette}{DISCBIO}(object, K) - -\S4method{plottSNE}{DISCBIO}(object) } \arguments{ \item{object}{\code{DISCBIO} class object.} From 6ea0900c41fc31498d5ca650597f4f79a2a48763 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 3 Aug 2020 16:11:18 +0200 Subject: [PATCH 85/99] Merged plottsneMB into plottSNE --- DESCRIPTION | 1 - NAMESPACE | 1 - R/DIscBIO-generic-clustheatmap.R | 4 ++- R/DIscBIO-generic-plottSNE.R | 34 +++++++++++++------ R/DIscBIO-generic-plottsneMB.R | 46 -------------------------- R/DIscBIO-generic-pseudoTimeOrdering.R | 9 +++-- man/plottSNE.Rd | 7 ++-- man/plottsneMB.Rd | 22 ------------ 8 files changed, 39 insertions(+), 85 deletions(-) delete mode 100644 R/DIscBIO-generic-plottsneMB.R delete mode 100644 man/plottsneMB.Rd diff --git a/DESCRIPTION b/DESCRIPTION index 93769f2..2401dff 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -78,7 +78,6 @@ Collate: 'DIscBIO-generic-plotSilhouette.R' 'DIscBIO-generic-plotSymbolstSNE.R' 'DIscBIO-generic-plottSNE.R' - 'DIscBIO-generic-plottsneMB.R' 'DIscBIO-generic-pseudoTimeOrdering.R' 'J48DT.R' 'J48DTeval.R' diff --git a/NAMESPACE b/NAMESPACE index 89ad442..2ead175 100644 --- a/NAMESPACE +++ b/NAMESPACE @@ -38,7 +38,6 @@ exportMethods(plotOrderTsne) exportMethods(plotSilhouette) exportMethods(plotSymbolstSNE) exportMethods(plottSNE) -exportMethods(plottsneMB) exportMethods(pseudoTimeOrdering) import(org.Hs.eg.db) importClassesFrom(SingleCellExperiment,SingleCellExperiment) diff --git a/R/DIscBIO-generic-clustheatmap.R b/R/DIscBIO-generic-clustheatmap.R index d71538e..cb26ad0 100644 --- a/R/DIscBIO-generic-clustheatmap.R +++ b/R/DIscBIO-generic-clustheatmap.R @@ -32,7 +32,9 @@ setGeneric( setMethod( "clustheatmap", signature = "DISCBIO", - definition = function(object, clustering_method, hmethod, plot) + definition = function( + object, clustering_method, hmethod, rseed, quiet, plot + ) { x <- object@fdata if (tolower(clustering_method) %in% c("k-means", "k")) { diff --git a/R/DIscBIO-generic-plottSNE.R b/R/DIscBIO-generic-plottSNE.R index 1fb9636..60d05ba 100644 --- a/R/DIscBIO-generic-plottSNE.R +++ b/R/DIscBIO-generic-plottSNE.R @@ -1,23 +1,37 @@ -#' @title tSNE map for K-means clustering -#' @description Visualizing the K-means clusters using tSNE maps +#' @title tSNE map +#' @description Visualizing the k-means or model-based clusters using tSNE maps #' @param object \code{DISCBIO} class object. #' @importFrom graphics text #' @return A plot of t-SNEs. setGeneric("plottSNE", function(object) standardGeneric("plottSNE")) -#' @rdname plotSilhouette +#' @rdname plottSNE #' @export setMethod( "plottSNE", signature = "DISCBIO", definition = function(object) { - if (length(object@tsne) == 0) - stop("run comptsne before plottsne") + # ====================================================================== + # Validating + # ====================================================================== + ran_k <- length(object@tsne) > 0 + ran_m <- length(object@MBtsne) > 0 + if (ran_k) { + part <- object@kmeans$kpart + x <- object@tsne + } else if (ran_m) { + part <- object@MBclusters$clusterid + x <- object@MBtsne + } else { + stop("run comptsne before plottSNE") + } + # ====================================================================== + # Plotting + # ====================================================================== col <- c("black", "blue", "green", "red", "yellow", "gray") - part <- object@kmeans$kpart plot( - object@tsne, + x, las = 1, xlab = "Dim 1", ylab = "Dim 2", @@ -25,11 +39,11 @@ setMethod( cex = 1.5, col = "lightgrey" ) - for (i in 1:max(part)) { + for (i in seq_len(part)) { if (sum(part == i) > 0) { text( - object@tsne[part == i, 1], - object@tsne[part == i, 2], + x[part == i, 1], + x[part == i, 2], i, col = col[i], cex = .75, diff --git a/R/DIscBIO-generic-plottsneMB.R b/R/DIscBIO-generic-plottsneMB.R deleted file mode 100644 index 00140be..0000000 --- a/R/DIscBIO-generic-plottsneMB.R +++ /dev/null @@ -1,46 +0,0 @@ -#' @title tSNE map for Model-based clustering -#' @description Visualizing the Model-based clusters using tSNE maps -#' @param object \code{DISCBIO} class object. -#' @param K A numeric value of the number of clusters -#' @importFrom graphics text -#' @return A plot of t-SNEs. -setGeneric( - name = "plottsneMB", - def = function(object, K = length(table(object@MBclusters$clusterid))) { - standardGeneric("plottsneMB") - } -) - -#' @export -#' @rdname plottsneMB -setMethod( - f = "plottsneMB", - signature = "DISCBIO", - definition = function(object, K) { - if (length(object@MBtsne) == 0) - stop("run comptsne before plottsneMB") - col = c("black", "blue", "green", "red", "yellow", "gray") - part <- object@MBclusters$clusterid - plot( - object@MBtsne, - xlab = "Dim 1", - ylab = "Dim 2", - pch = 20, - cex = 1.5, - col = "lightgrey", - las = 1 - ) - for (i in 1:K) { - if (sum(part == i) > 0) { - text( - object@MBtsne[part == i, 1], - object@MBtsne[part == i, 2], - i, - col = col[i], - cex = .75, - font = 4 - ) - } - } - } -) \ No newline at end of file diff --git a/R/DIscBIO-generic-pseudoTimeOrdering.R b/R/DIscBIO-generic-pseudoTimeOrdering.R index 2be44e3..bca289e 100644 --- a/R/DIscBIO-generic-pseudoTimeOrdering.R +++ b/R/DIscBIO-generic-pseudoTimeOrdering.R @@ -31,17 +31,22 @@ setMethod( ran_k <- length(object@kmeans$kpart) > 0 ran_m <- length(object@MBclusters) > 0 if (ran_k) { + Obj <- object@fdata Names <- object@cpart - lpsmclust <- Exprmclust(Obj, K = 4, reduce = F, cluster = Clusters) + lpsmclust <- Exprmclust(Obj, K = 4, reduce = F, cluster = Names) lpsorder <- TSCANorder(lpsmclust) } else if (ran_m) { + Obj <- object@fdata Names <- names(object@MBclusters$clusterid) lpsmclust <- object@MBclusters lpsorder <- TSCANorder(lpsmclust) } else { stop("run clustexp before this pseudoTimeOrdering") } - sampleNames <- colnames(object@fdata) + # ====================================================================== + # Ordering + # ====================================================================== + sampleNames <- colnames(Obj) orderID <- lpsorder order <- c(1:length(lpsorder)) orderTable <- data.frame(order, orderID) diff --git a/man/plottSNE.Rd b/man/plottSNE.Rd index 6300ef1..a69f68d 100644 --- a/man/plottSNE.Rd +++ b/man/plottSNE.Rd @@ -2,9 +2,12 @@ % Please edit documentation in R/DIscBIO-generic-plottSNE.R \name{plottSNE} \alias{plottSNE} -\title{tSNE map for K-means clustering} +\alias{plottSNE,DISCBIO-method} +\title{tSNE map} \usage{ plottSNE(object) + +\S4method{plottSNE}{DISCBIO}(object) } \arguments{ \item{object}{\code{DISCBIO} class object.} @@ -13,5 +16,5 @@ plottSNE(object) A plot of t-SNEs. } \description{ -Visualizing the K-means clusters using tSNE maps +Visualizing the k-means or model-based clusters using tSNE maps } diff --git a/man/plottsneMB.Rd b/man/plottsneMB.Rd deleted file mode 100644 index 446a3db..0000000 --- a/man/plottsneMB.Rd +++ /dev/null @@ -1,22 +0,0 @@ -% Generated by roxygen2: do not edit by hand -% Please edit documentation in R/DIscBIO-generic-plottsneMB.R -\name{plottsneMB} -\alias{plottsneMB} -\alias{plottsneMB,DISCBIO-method} -\title{tSNE map for Model-based clustering} -\usage{ -plottsneMB(object, K = length(table(object@MBclusters$clusterid))) - -\S4method{plottsneMB}{DISCBIO}(object, K = length(table(object@MBclusters$clusterid))) -} -\arguments{ -\item{object}{\code{DISCBIO} class object.} - -\item{K}{A numeric value of the number of clusters} -} -\value{ -A plot of t-SNEs. -} -\description{ -Visualizing the Model-based clusters using tSNE maps -} From b1fbb32775fe0b0b6a76e1c34d071eaa57da9ba0 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 3 Aug 2020 16:23:34 +0200 Subject: [PATCH 86/99] Changed Travis checks from Bioconductor --- .travis.yml | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/.travis.yml b/.travis.yml index 769754d..5a35725 100644 --- a/.travis.yml +++ b/.travis.yml @@ -5,5 +5,5 @@ cache: packages: true timeout: 3000 r: - - bioc-release - - bioc-devel \ No newline at end of file + - release + - devel \ No newline at end of file From 6e489a6a978be2836327f5cf51c057bf886b1b9f Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 3 Aug 2020 16:42:06 +0200 Subject: [PATCH 87/99] Reduced dataset --- .Rbuildignore | 4 +-- R/customConverters.R | 1 - R/datasets.R | 20 ++--------- man/valuesG1msReduced_treated_K.Rd | 9 ----- man/valuesG1msReduced_treated_MB.Rd | 9 ----- man/{valuesG1msRed.Rd => valuesG1msTest.Rd} | 8 ++--- tests/testthat/test-DIscBIO.IMP.R | 40 ++++++--------------- 7 files changed, 19 insertions(+), 72 deletions(-) delete mode 100644 man/valuesG1msReduced_treated_K.Rd delete mode 100644 man/valuesG1msReduced_treated_MB.Rd rename man/{valuesG1msRed.Rd => valuesG1msTest.Rd} (82%) diff --git a/.Rbuildignore b/.Rbuildignore index 485cdd7..54840d8 100644 --- a/.Rbuildignore +++ b/.Rbuildignore @@ -6,8 +6,8 @@ CONTRIBUTING.md DIscBIOlogo.png README.md notebook -data/valuesG1msReduced -data/valuesG1msTest +data/valuesG1msReduced.rda +data/valuesG1msRed.rda data/valuesG1ms.rda data/genes20k.rda data/pan_indrop_matrix_8000cells_18556genes.rda diff --git a/R/customConverters.R b/R/customConverters.R index fa36afe..bf7b5ab 100644 --- a/R/customConverters.R +++ b/R/customConverters.R @@ -25,7 +25,6 @@ customConvertFeats <- function(x, verbose = TRUE) { # uncomment myDict <- DIscBIO::HumanMouseGeneIds - # myDict <- HumanMouseGeneIds myDict <- myDict[!is.na(myDict[, "ENSEMBL"]), ] # diff --git a/R/datasets.R b/R/datasets.R index 3a6b4c6..00391e4 100644 --- a/R/datasets.R +++ b/R/datasets.R @@ -1,12 +1,12 @@ #' @title Single-cells data from a myxoid liposarcoma cell line #' -#' @description A dataset of 30 single cells from a myxoid liposarcoma cell -#' line. Columns refer to samples and rows refer to genes. The last 92 rows +#' @description A sample of single cells from a myxoid liposarcoma cell +#' line. Columns refer to samples and rows refer to genes. The last rows #' refer to external RNA controls consortium (ERCC) spike-ins. This dataset is #' part of a larger dataset containing 94 single cells. The complete dataset #' is fully compatible with this package and an rda file can be obtained at #' https://github.com/ocbe-uio/DIscBIO/blob/dev/data/valuesG1ms.rda -#' @name valuesG1msRed +#' @name valuesG1msTest #' @docType data NULL @@ -20,18 +20,4 @@ NULL #' BioConductor libraries. #' @name HumanMouseGeneIds #' @docType data -NULL - -#' @title Single-cells data from a myxoid liposarcoma cell line (K variant) -#' -#' @description Equivalent to valuesG1msReduced after treatment and K-means clustering. -#' @name valuesG1msReduced_treated_K -#' @docType data -NULL - -#' @title Single-cells data from a myxoid liposarcoma cell line (MB variant) -#' -#' @description Equivalent to valuesG1msReduced after treatment and model-based clustering. -#' @name valuesG1msReduced_treated_MB -#' @docType data NULL \ No newline at end of file diff --git a/man/valuesG1msReduced_treated_K.Rd b/man/valuesG1msReduced_treated_K.Rd deleted file mode 100644 index a6c744a..0000000 --- a/man/valuesG1msReduced_treated_K.Rd +++ /dev/null @@ -1,9 +0,0 @@ -% Generated by roxygen2: do not edit by hand -% Please edit documentation in R/datasets.R -\docType{data} -\name{valuesG1msReduced_treated_K} -\alias{valuesG1msReduced_treated_K} -\title{Single-cells data from a myxoid liposarcoma cell line (K variant)} -\description{ -Equivalent to valuesG1msReduced after treatment and K-means clustering. -} diff --git a/man/valuesG1msReduced_treated_MB.Rd b/man/valuesG1msReduced_treated_MB.Rd deleted file mode 100644 index 2d28a20..0000000 --- a/man/valuesG1msReduced_treated_MB.Rd +++ /dev/null @@ -1,9 +0,0 @@ -% Generated by roxygen2: do not edit by hand -% Please edit documentation in R/datasets.R -\docType{data} -\name{valuesG1msReduced_treated_MB} -\alias{valuesG1msReduced_treated_MB} -\title{Single-cells data from a myxoid liposarcoma cell line (MB variant)} -\description{ -Equivalent to valuesG1msReduced after treatment and model-based clustering. -} diff --git a/man/valuesG1msRed.Rd b/man/valuesG1msTest.Rd similarity index 82% rename from man/valuesG1msRed.Rd rename to man/valuesG1msTest.Rd index 263d4b2..5249a1d 100644 --- a/man/valuesG1msRed.Rd +++ b/man/valuesG1msTest.Rd @@ -1,12 +1,12 @@ % Generated by roxygen2: do not edit by hand % Please edit documentation in R/datasets.R \docType{data} -\name{valuesG1msRed} -\alias{valuesG1msRed} +\name{valuesG1msTest} +\alias{valuesG1msTest} \title{Single-cells data from a myxoid liposarcoma cell line} \description{ -A dataset of 30 single cells from a myxoid liposarcoma cell - line. Columns refer to samples and rows refer to genes. The last 92 rows +A sample of single cells from a myxoid liposarcoma cell + line. Columns refer to samples and rows refer to genes. The last rows refer to external RNA controls consortium (ERCC) spike-ins. This dataset is part of a larger dataset containing 94 single cells. The complete dataset is fully compatible with this package and an rda file can be obtained at diff --git a/tests/testthat/test-DIscBIO.IMP.R b/tests/testthat/test-DIscBIO.IMP.R index 1a7d219..4e9daa6 100644 --- a/tests/testthat/test-DIscBIO.IMP.R +++ b/tests/testthat/test-DIscBIO.IMP.R @@ -4,10 +4,10 @@ context("Data loading and pre-processing") -sc <- DISCBIO(valuesG1msRed) # Reduced dataset used for testing +sc <- DISCBIO(valuesG1msTest) # Reduced dataset used for testing test_that("Loading datasets generate the expected output", { - expect_equal(dim(valuesG1msRed), c(1000, 21)) + expect_equal(dim(valuesG1msTest), c(800, 15)) }) test_that("Data signature changes", { @@ -19,7 +19,7 @@ test_that("Data signature changes", { sc <- NoiseFiltering(sc, plot=FALSE, export=FALSE, quiet=TRUE) test_that("Noise filtering is added", { - expect_equal(length(sc@noiseF), 272) + expect_equal(length(sc@noiseF), 163) }) # In this case this function is used to normalize the reads @@ -30,14 +30,14 @@ sc <- Normalizedata( test_that("Data is normalized", { expect_equal(class(sc@fdata), "data.frame") - expect_output(str(sc@fdata), "908 obs. of 21 variables") + expect_output(str(sc@fdata), "708 obs. of 15 variables") }) # This function can be used for: 1- filtering and normalizing the dataset that has no ERCC. 2- to normalize and filter genes and cells after the noise filtering. sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE, quiet=TRUE) test_that("Data is normalized", { - expect_equal(dim(sc@fdata), c(272, 21)) + expect_equal(dim(sc@fdata), c(163, 15)) }) # ---------------------------------------------------------------------------- # @@ -51,13 +51,13 @@ sc <- comptSNE(sc, rseed=15555, quiet=TRUE, max_iter=1, epoch=10) test_that("tSNE is computed", { expect_equal(class(sc@tsne), "data.frame") - expect_output(str(sc@tsne), "21 obs. of 2 variables") + expect_output(str(sc@tsne), "15 obs. of 2 variables") }) test_that("Cluster plots output is as expexted", { expect_equivalent( object = Jaccard(sc, Clustering="K-means", K=2, plot=FALSE, R=5), - expected = c(.327, .425), + expected = c(.417, .413), tolerance = .01 ) expect_equal( @@ -77,31 +77,11 @@ Outliers <- FindOutliers( Order <- KmeanOrder(sc, quiet = TRUE, export = FALSE) test_that("Outliers are the expected", { - expect_equivalent(Outliers, c(3, 7, 14)) + expect_equivalent(Outliers, c(3, 10)) expect_equivalent( object = Order@kordering, expected = c( - 15, 13, 5, 14, 20, 18, 16, 21, 17, 9, 8, 12, 3, 10, 2, 11, 7, 19, - 6, 4, 1 + 5, 6, 13, 2, 3, 1, 4, 7, 12, 9, 15, 8, 10, 11, 14 ) ) -}) - -# --------------------- Differential Expression Analysis --------------------- # - -context("Differential Expression Analysis") - -# differential expression analysis between two particular clusters. -cdiff3 <- suppressMessages( - DEGanalysis2clust( - sc, Clustering="K-means", K=2, fdr=.2, name="Name", First="CL1", - Second="CL2", export=FALSE, quiet=TRUE, plot=FALSE, nperms=2, nresamp=2 - ) -) - -test_that("DEGs are calculated", { - expect_equivalent( - object = sapply(cdiff3, function(x) class(x)[1]), - expected = c("matrix", "data.frame", "data.frame", "data.frame") - ) -}) +}) \ No newline at end of file From 790ab63cee9091f230e6466b53fb6b7e55414c26 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 3 Aug 2020 17:07:48 +0200 Subject: [PATCH 88/99] Eliminated unused column from dictionary --- data/HumanMouseGeneIds.rda | Bin 388328 -> 386544 bytes 1 file changed, 0 insertions(+), 0 deletions(-) diff --git a/data/HumanMouseGeneIds.rda b/data/HumanMouseGeneIds.rda index 7d0a6c8b79f16b1d8bf39088ef1490033c7d0af4..4e4c1e7757ee7bc208c98605ee5b03dcf69427ab 100644 GIT binary patch literal 386544 zcmV(lK=i-;H+ooF0004LBHlIv03iV!0000G&sfauOZxBrT>vQ&2UKVgRpfklI z#ix#GmMP!ctZhp*SntSRl*d1=kl*bbFQE%3&1WMDko05WwVZ%-{B&I-?JgfXE%Y*b z(W#HC>lf+9a?2o7>Kb<}=jl$CAk_VBzC;;r2 z@l618;Dgs$C2X#y-wQcmyzKUcsv^eLk}8rNnvc4Q{su=GF;57_!Pno~2GcmUP>y_9 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z646DvP<(q6>OmH<>Ons?dIg{21oFS$FdnXF?@Rg`Dj;8ERx;dQr1@8o8rWeBCeq-H zaE;p8ye=CwM7!+a3r2KnSWJ;U2X)Z+FtjffiZph-A2ok4HO`KCgokUMw)_9a`ng3j zZ0g}%H*aRfb(NFoJ~spvrT^ZjMSlNlnTk5Wy~S)?pD!zz71VfyQhY|8n3bDiDep4? z1>2bAZWQLd(cS%Mp4z}4ydRj+qt_^THF2?3-%7d`ctKwh1a{uga>DX#Pxb==oxIsUsvn$oEzB+nH71u#($DrJ>*eLtf})VbpAig2kb#zpM*7 zOAXk6*&^*W;uWNq_A~8~V?6|hkuVQcZuq?JV>=%`=l3@C&p154G;mgpc>`Q z-^=<0ghl5YnL~{B(eix4u3IDtK4r4}*|HIwov9>(BtL=~yS#P>lKLZQbNq6gdg;n0 zb52`LT5m@z{bCL&di>J+bLR|CIRwq@+zVrh&9yOzEZWqsCClBX zn#d;?`t=jYzZvj6J6+PM&Gt^v_~*^t93wH9jp`SqgnXNpbVP;!yc_^}1NF}fD$gj^o?!-r+Is!M0T3ALLgTu{w^ngz()7FGNny0eS#~4w z!lMT}h1P%TzVD;LPhaOCKnc9TGGK54?R0%Dr_NLdKH)2O{$-yGAOmGjFmMroIKjZ* zsi@kPr1R53LG<>lt08TN{i08<3~97s#_RU#7y$i01BU Date: Mon, 3 Aug 2020 17:27:32 +0200 Subject: [PATCH 89/99] Removed some suggested packages --- DESCRIPTION | 3 --- 1 file changed, 3 deletions(-) diff --git a/DESCRIPTION b/DESCRIPTION index 2401dff..b85a7a8 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -44,10 +44,7 @@ Imports: methods, TSCAN, boot, httr, mclust, statmod, igraph, Depends: R (>= 4.0), SingleCellExperiment Suggests: - knitr, - rmarkdown, testthat, - enrichR, Seurat, BiocManager LazyData: true From d56e5ca95b2bdd08d00ed0d740ccb67144b0c293 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 3 Aug 2020 17:53:57 +0200 Subject: [PATCH 90/99] Further reduction in suggested packages --- DESCRIPTION | 3 +-- 1 file changed, 1 insertion(+), 2 deletions(-) diff --git a/DESCRIPTION b/DESCRIPTION index b85a7a8..d957fc6 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -45,8 +45,7 @@ Depends: R (>= 4.0), SingleCellExperiment Suggests: testthat, - Seurat, - BiocManager + Seurat LazyData: true RoxygenNote: 7.1.1 URL: https://github.com/ocbe-uio/DIscBIO#usage From 04f83c8aad3e6d31115186affeafaef252472bb4 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Mon, 3 Aug 2020 17:57:37 +0200 Subject: [PATCH 91/99] Removed outdated install script --- install.R | 18 ------------------ 1 file changed, 18 deletions(-) delete mode 100644 install.R diff --git a/install.R b/install.R deleted file mode 100644 index 0fb2dd3..0000000 --- a/install.R +++ /dev/null @@ -1,18 +0,0 @@ -if (!requireNamespace("BiocManager", quietly = TRUE)) install.packages("BiocManager") - -BiocManager::install( - c( - "pheatmap", "cluster", "mclust", "flexmix", - "lattice", "fpc", "amap", "RColorBrewer", "locfit", "TSCAN", - "genefilter", "statmod", "ggplot2", "gplots", "DESeq2", - "matrixStats", "robustbase", "philentropy", "igraph", "boot", - "biomaRt", "tidyr", "calibrate", "partykit", "RWeka", "rpart", - "rpart.plot", "imager", "png", "NetIndices", "httr", "jsonlite", - "tidyverse", "samr", "tidyverse", "org.Hs.eg.db", "AnnotationDbi", - "enrichR", "tsne" - ) -) - -# BiocInstaller appears to be needed to install DIscBIO from source -# it shouldn't be needed, however -install.packages("BiocInstaller", repos="http://bioconductor.org/packages/2.13/bioc") From fac5aee33f35dc173149f76ff3f5a6c530498651 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Tue, 25 Aug 2020 10:41:11 +0200 Subject: [PATCH 92/99] Reinstated examples --- R/DIscBIO-classes.R | 7 +++++++ R/DIscBIO-generic-ClustDiffGenes.R | 8 +++++++- R/DIscBIO-generic-Exprmclust.R | 11 +++++++++++ R/DIscBIO-generic-FinalPreprocessing.R | 5 +++++ R/DIscBIO-generic-FindOutliers.R | 4 ++++ R/DIscBIO-generic-NoiseFiltering.R | 5 +++++ R/DIscBIO-generic-Normalizedata.R | 10 ++++++++++ R/DIscBIO-generic-PCAplotSymbols.R | 10 ++++++++++ R/DIscBIO-generic-comptSNE.R | 5 +++++ R/customConverters.R | 6 ++++++ 10 files changed, 70 insertions(+), 1 deletion(-) diff --git a/R/DIscBIO-classes.R b/R/DIscBIO-classes.R index db2da5a..c5ca4a0 100644 --- a/R/DIscBIO-classes.R +++ b/R/DIscBIO-classes.R @@ -51,6 +51,13 @@ #' #' @importClassesFrom SingleCellExperiment SingleCellExperiment #' @export +#' +#' @examples +#' class(valuesG1msTest) +#' G1_reclassified <- DISCBIO(valuesG1msTest) +#' class(G1_reclassified) +#' str(G1_reclassified, max.level=2) +#' identical(G1_reclassified@expdataAll, valuesG1msTest) DISCBIO <- setClass( Class = "DISCBIO", slots = c( diff --git a/R/DIscBIO-generic-ClustDiffGenes.R b/R/DIscBIO-generic-ClustDiffGenes.R index af757d1..4df2e6e 100644 --- a/R/DIscBIO-generic-ClustDiffGenes.R +++ b/R/DIscBIO-generic-ClustDiffGenes.R @@ -16,7 +16,13 @@ #' @rdname ClustDiffGenes #' @return A list containing two tables. #' @export -#' +#' @examples +#' sc <- DISCBIO(valuesG1msTest) +#' sc <- Clustexp(sc, cln=3, quiet=TRUE) +#' cdiff <- ClustDiffGenes(sc, K=3, fdr=.3, export=FALSE) +#' str(cdiff) +#' cdiff[[2]] + setGeneric( "ClustDiffGenes", function( diff --git a/R/DIscBIO-generic-Exprmclust.R b/R/DIscBIO-generic-Exprmclust.R index e381249..f19404d 100644 --- a/R/DIscBIO-generic-Exprmclust.R +++ b/R/DIscBIO-generic-Exprmclust.R @@ -18,6 +18,17 @@ #' the MBclusters slot filled. If the `object` is a data frame, the function #' returns a named list containing the four objects that together correspond #' to the contents of the MBclusters slot. +#' @examples +#' sc <- DISCBIO(valuesG1msTest) +#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE, plot=FALSE) +#' sc <- Normalizedata( +#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, +#' dsn=1, rseed=17000 +#' ) +#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) +#' sc <- Exprmclust(sc, reduce=FALSE, K=1) +#' print(sc@MBclusters) +#' setGeneric( name = "Exprmclust", def = function(object, diff --git a/R/DIscBIO-generic-FinalPreprocessing.R b/R/DIscBIO-generic-FinalPreprocessing.R index 34178b2..0b6bb18 100644 --- a/R/DIscBIO-generic-FinalPreprocessing.R +++ b/R/DIscBIO-generic-FinalPreprocessing.R @@ -8,6 +8,11 @@ #' @param quiet if `TRUE`, intermediary output is suppressed #' @param fileName File name for exporting (if `export = TRUE`) #' @return The DISCBIO-class object input with the FinalGeneList slot filled. +#' @examples +#' sc <- DISCBIO(valuesG1msTest) +#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) +#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) +#' setGeneric( "FinalPreprocessing", function( diff --git a/R/DIscBIO-generic-FindOutliers.R b/R/DIscBIO-generic-FindOutliers.R index 8c0c51f..5785602 100644 --- a/R/DIscBIO-generic-FindOutliers.R +++ b/R/DIscBIO-generic-FindOutliers.R @@ -21,6 +21,10 @@ #' @importFrom stats coef pnbinom #' @return A named vector of the genes containing outlying cells and the number #' of cells on each. +#' @examples +#' sc <- DISCBIO(valuesG1msTest) +#' sc <- Clustexp(sc, cln=2) # K-means clustering +#' FindOutliers(sc, K=2) #' setGeneric( "FindOutliers", diff --git a/R/DIscBIO-generic-NoiseFiltering.R b/R/DIscBIO-generic-NoiseFiltering.R index 0add7d0..2f34189 100644 --- a/R/DIscBIO-generic-NoiseFiltering.R +++ b/R/DIscBIO-generic-NoiseFiltering.R @@ -25,6 +25,11 @@ #' @importFrom statmod glmgam.fit #' @note This function should be used only if the dataset has ERCC. #' @return The DISCBIO-class object input with the noiseF slot filled. +#' @examples +#' sc <- DISCBIO(valuesG1msTest) # changes signature of data +#' sd_filtered <- NoiseFiltering(sc, export=FALSE) +#' str(sd_filtered) +#' setGeneric( name = "NoiseFiltering", def = function( diff --git a/R/DIscBIO-generic-Normalizedata.R b/R/DIscBIO-generic-Normalizedata.R index 4357069..875b86f 100644 --- a/R/DIscBIO-generic-Normalizedata.R +++ b/R/DIscBIO-generic-Normalizedata.R @@ -23,6 +23,16 @@ #' results #' @include DIscBIO-classes.R #' @return The DISCBIO-class object input with the ndata and fdata slots filled. +#' @examples +#' sc <- DISCBIO(valuesG1msTest) # changes signature of data +#' +#' # In this case this function is used to normalize the reads +#' sc_normal <- Normalizedata( +#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, +#' dsn=1, rseed=17000 +#' ) +#' summary(sc_normal@fdata) +#' setGeneric( "Normalizedata", function( diff --git a/R/DIscBIO-generic-PCAplotSymbols.R b/R/DIscBIO-generic-PCAplotSymbols.R index 0de0caf..52b068c 100644 --- a/R/DIscBIO-generic-PCAplotSymbols.R +++ b/R/DIscBIO-generic-PCAplotSymbols.R @@ -6,6 +6,16 @@ #' @importFrom grDevices rainbow #' @importFrom graphics legend #' @return Plot of the Principal Components +#' @examples +#' sc <- DISCBIO(valuesG1msRed) +#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) +#' sc <- Normalizedata( +#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, +#' dsn=1, rseed=17000 +#' ) +#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) +#' sc <- Exprmclust(sc, K=2) +#' PCAplotSymbols(sc) #' setGeneric("PCAplotSymbols", function(object, types = NULL) standardGeneric("PCAplotSymbols")) diff --git a/R/DIscBIO-generic-comptSNE.R b/R/DIscBIO-generic-comptSNE.R index acb7a34..e6740a3 100644 --- a/R/DIscBIO-generic-comptSNE.R +++ b/R/DIscBIO-generic-comptSNE.R @@ -11,6 +11,11 @@ #' @importFrom tsne tsne #' @importFrom stats as.dist cor #' @return The DISCBIO-class object input with the tsne slot filled. +#' @examples +#' sc <- DISCBIO(valuesG1msTest) # changes signature of data +#' sc <- Clustexp(sc, cln=2) # data must be clustered before plottin +#' sc <- comptSNE(sc, max_iter=30) +#' head(sc@tsne) #' setGeneric( name = "comptSNE", diff --git a/R/customConverters.R b/R/customConverters.R index bf7b5ab..ac1e328 100644 --- a/R/customConverters.R +++ b/R/customConverters.R @@ -147,6 +147,12 @@ as.DISCBIO <- function(x, ...) { #' @return a SingleCellExperiment-class object #' #' @export +#' @examples +#' g1_disc <- DISCBIO(valuesG1msTest) +#' class(g1_disc) +#' g1_sce <- DISCBIO2SingleCellExperiment(g1_disc) +#' class(g1_sce) +#' #' DISCBIO2SingleCellExperiment <- function(x) { return(x@SingleCellExperiment) From b29c423bc47e5bd593289a2b18238d9d4a387906 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Tue, 25 Aug 2020 10:41:32 +0200 Subject: [PATCH 93/99] Changed default argument value --- R/DIscBIO-generic-Clustexp.R | 5 ++++- 1 file changed, 4 insertions(+), 1 deletion(-) diff --git a/R/DIscBIO-generic-Clustexp.R b/R/DIscBIO-generic-Clustexp.R index 1b90ace..e4967a6 100644 --- a/R/DIscBIO-generic-Clustexp.R +++ b/R/DIscBIO-generic-Clustexp.R @@ -30,7 +30,10 @@ #' @importFrom graphics pairs #' @importFrom methods is #' @return The DISCBIO-class object input with the cpart slot filled. -setGeneric("Clustexp", function(object, clustnr = 20, bootnr = 50, +#' @examples +#' sc <- DISCBIO(valuesG1msTest) # changes signature of data +#' sc <- Clustexp(sc, cln=2) +setGeneric("Clustexp", function(object, clustnr = 3, bootnr = 50, metric = "pearson", do.gap = TRUE, SE.method = "Tibs2001SEmax", SE.factor = .25, B.gap = 50, cln = 0, rseed = NULL, quiet = FALSE) { From 92ebfb405cae072fc50b140412112a76b860006c Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Tue, 25 Aug 2020 10:41:46 +0200 Subject: [PATCH 94/99] Improved documentation, formatting --- R/DIscBIO-generic-ClustDiffGenes.R | 2 +- R/DIscBIO-generic-Exprmclust.R | 135 ++++++++++++++-------------- man/ClustDiffGenes.Rd | 9 +- man/Clustexp.Rd | 8 +- man/DISCBIO.Rd | 7 ++ man/DISCBIO2SingleCellExperiment.Rd | 8 ++ man/FinalPreprocessing.Rd | 6 ++ man/FindOutliers.Rd | 6 ++ man/NoiseFiltering.Rd | 6 ++ man/Normalizedata.Rd | 11 +++ man/PCAplotSymbols.Rd | 12 +++ man/comptSNE.Rd | 7 ++ 12 files changed, 145 insertions(+), 72 deletions(-) diff --git a/R/DIscBIO-generic-ClustDiffGenes.R b/R/DIscBIO-generic-ClustDiffGenes.R index 4df2e6e..5b2e4cc 100644 --- a/R/DIscBIO-generic-ClustDiffGenes.R +++ b/R/DIscBIO-generic-ClustDiffGenes.R @@ -1,5 +1,5 @@ #' @title ClustDiffGenes -#' @description description +#' @description Creates a table of cluster differences #' @param object \code{DISCBIO} class object. #' @param K A numeric value of the number of clusters. #' @param pValue A numeric value of the p-value. Default is 0.05. diff --git a/R/DIscBIO-generic-Exprmclust.R b/R/DIscBIO-generic-Exprmclust.R index f19404d..e28a435 100644 --- a/R/DIscBIO-generic-Exprmclust.R +++ b/R/DIscBIO-generic-Exprmclust.R @@ -31,80 +31,79 @@ #' setGeneric( name = "Exprmclust", -def = function(object, - K = 3, - modelNames = "VVV", - reduce = TRUE, - cluster = NULL, - quiet = FALSE) -standardGeneric("Exprmclust") +def = function( + object, + K = 3, + modelNames = "VVV", + reduce = TRUE, + cluster = NULL, + quiet = FALSE +) { + standardGeneric("Exprmclust") +} ) #' @export #' @rdname Exprmclust setMethod( -f = "Exprmclust", -signature = "DISCBIO", -definition = function(object, - K = 3, - modelNames = "VVV", - reduce = TRUE, - cluster = NULL, - quiet = FALSE) { - obj <- object@fdata - if (reduce) { - sdev <- prcomp(t(obj), scale = T)$sdev[1:20] - x <- 1:20 - optpoint <- which.min(sapply(2:10, function(i) { - x2 <- pmax(0, x - i) - sum(lm(sdev ~ x + x2)$residuals ^ 2) - })) - pcadim = optpoint + 1 - tmpdata <- t(apply(obj, 1, scale)) - colnames(tmpdata) <- colnames(obj) - tmppc <- prcomp(t(tmpdata), scale = T) - pcareduceres <- - t(tmpdata) %*% tmppc$rotation[, 1:pcadim] - } - else { - pcareduceres <- t(obj) - } - if (is.null(cluster)) { - K <- K[K > 1] - res <- Mclust( - data = pcareduceres, - G = K, - modelNames = modelNames, - warn = FALSE, - verbose = !quiet - ) - if (is.null(res)) stop("Unable to cluster. Try a lower value for K.") - clusterid <- apply(res$z, 1, which.max) - clunum <- res$G - } else { - clunum <- length(unique(cluster)) - clusterid <- cluster - } - clucenter <- - matrix(0, ncol = ncol(pcareduceres), nrow = clunum) - for (cid in 1:clunum) { - clucenter[cid, ] <- colMeans( - pcareduceres[names(clusterid[clusterid == cid]), , drop = FALSE] - ) - } - dp <- as.matrix(dist(clucenter)) - gp <- - graph.adjacency(dp, mode = "undirected", weighted = TRUE) - dp_mst <- minimum.spanning.tree(gp) - full_List <- - list( - pcareduceres = pcareduceres, - MSTtree = dp_mst, - clusterid = clusterid, - clucenter = clucenter + f = "Exprmclust", + signature = "DISCBIO", + definition = function(object, K, modelNames, reduce, cluster, quiet) { + obj <- object@fdata + if (reduce) { + sdev <- prcomp(t(obj), scale = T)$sdev[1:20] + x <- 1:20 + optpoint <- which.min( + sapply( + 2:10, + function(i) { + x2 <- pmax(0, x - i) + sum(lm(sdev ~ x + x2)$residuals ^ 2) + } + ) + ) + pcadim <- optpoint + 1 + tmpdata <- t(apply(obj, 1, scale)) + colnames(tmpdata) <- colnames(obj) + tmppc <- prcomp(t(tmpdata), scale = T) + pcareduceres <- t(tmpdata) %*% tmppc$rotation[, 1:pcadim] + } + else { + pcareduceres <- t(obj) + } + if (is.null(cluster)) { + K <- K[K > 1] + res <- Mclust( + data = pcareduceres, + G = K, + modelNames = modelNames, + warn = FALSE, + verbose = !quiet + ) + if (is.null(res)) stop("Unable to cluster. Try a lower value for K.") + clusterid <- apply(res$z, 1, which.max) + clunum <- res$G + } else { + clunum <- length(unique(cluster)) + clusterid <- cluster + } + clucenter <- matrix(0, ncol = ncol(pcareduceres), nrow = clunum) + for (cid in 1:clunum) { + clucenter[cid, ] <- colMeans( + pcareduceres[names(clusterid[clusterid == cid]), , drop = FALSE] + ) + } + dp <- as.matrix(dist(clucenter)) + gp <- graph.adjacency(dp, mode = "undirected", weighted = TRUE) + dp_mst <- minimum.spanning.tree(gp) + full_List <- list( + pcareduceres = pcareduceres, + MSTtree = dp_mst, + clusterid = clusterid, + clucenter = clucenter ) - object@MBclusters <- full_List - return(object) + object@MBclusters <- full_List + return(object) } ) diff --git a/man/ClustDiffGenes.Rd b/man/ClustDiffGenes.Rd index a98aa4f..ff1bb8a 100644 --- a/man/ClustDiffGenes.Rd +++ b/man/ClustDiffGenes.Rd @@ -57,5 +57,12 @@ excel file. Default is TRUE.} A list containing two tables. } \description{ -description +Creates a table of cluster differences +} +\examples{ +sc <- DISCBIO(valuesG1msTest) +sc <- Clustexp(sc, cln=3, quiet=TRUE) +cdiff <- ClustDiffGenes(sc, K=3, fdr=.3, export=FALSE) +str(cdiff) +cdiff[[2]] } diff --git a/man/Clustexp.Rd b/man/Clustexp.Rd index 7711797..c5874ac 100644 --- a/man/Clustexp.Rd +++ b/man/Clustexp.Rd @@ -8,7 +8,7 @@ \usage{ Clustexp( object, - clustnr = 20, + clustnr = 3, bootnr = 50, metric = "pearson", do.gap = TRUE, @@ -22,7 +22,7 @@ Clustexp( \S4method{Clustexp}{DISCBIO}( object, - clustnr = 20, + clustnr = 3, bootnr = 50, metric = "pearson", do.gap = TRUE, @@ -76,3 +76,7 @@ The DISCBIO-class object input with the cpart slot filled. This functions performs the initial clustering of the RaceID algorithm. } +\examples{ +sc <- DISCBIO(valuesG1msTest) # changes signature of data +sc <- Clustexp(sc, cln=2) +} diff --git a/man/DISCBIO.Rd b/man/DISCBIO.Rd index 261b1b2..e3d863b 100644 --- a/man/DISCBIO.Rd +++ b/man/DISCBIO.Rd @@ -82,3 +82,10 @@ noise filtering.} from running the noise filtering or/and the expression filtering.} }} +\examples{ +class(valuesG1msTest) +G1_reclassified <- DISCBIO(valuesG1msTest) +class(G1_reclassified) +str(G1_reclassified, max.level=2) +identical(G1_reclassified@expdataAll, valuesG1msTest) +} diff --git a/man/DISCBIO2SingleCellExperiment.Rd b/man/DISCBIO2SingleCellExperiment.Rd index adad853..16a96a6 100644 --- a/man/DISCBIO2SingleCellExperiment.Rd +++ b/man/DISCBIO2SingleCellExperiment.Rd @@ -16,3 +16,11 @@ a SingleCellExperiment-class object Extract the SingleCellExperiment input data from the corresponding input slot in a DISCBIO-class object } +\examples{ +g1_disc <- DISCBIO(valuesG1msTest) +class(g1_disc) +g1_sce <- DISCBIO2SingleCellExperiment(g1_disc) +class(g1_sce) + + +} diff --git a/man/FinalPreprocessing.Rd b/man/FinalPreprocessing.Rd index e6506c8..68af9d1 100644 --- a/man/FinalPreprocessing.Rd +++ b/man/FinalPreprocessing.Rd @@ -40,3 +40,9 @@ The DISCBIO-class object input with the FinalGeneList slot filled. \description{ This function generates the final filtered normalized dataset. } +\examples{ +sc <- DISCBIO(valuesG1msTest) +sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) +sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) + +} diff --git a/man/FindOutliers.Rd b/man/FindOutliers.Rd index 2d9913e..11a3074 100644 --- a/man/FindOutliers.Rd +++ b/man/FindOutliers.Rd @@ -64,3 +64,9 @@ A named vector of the genes containing outlying cells and the number \description{ This functions performs the outlier identification for k-means and model-based clustering } +\examples{ +sc <- DISCBIO(valuesG1msTest) +sc <- Clustexp(sc, cln=2) # K-means clustering +FindOutliers(sc, K=2) + +} diff --git a/man/NoiseFiltering.Rd b/man/NoiseFiltering.Rd index b043910..c22bb6f 100644 --- a/man/NoiseFiltering.Rd +++ b/man/NoiseFiltering.Rd @@ -74,3 +74,9 @@ Given a matrix or data frame of count data, this function \note{ This function should be used only if the dataset has ERCC. } +\examples{ +sc <- DISCBIO(valuesG1msTest) # changes signature of data +sd_filtered <- NoiseFiltering(sc, export=FALSE) +str(sd_filtered) + +} diff --git a/man/Normalizedata.Rd b/man/Normalizedata.Rd index 3c7e8dc..2e1fbca 100644 --- a/man/Normalizedata.Rd +++ b/man/Normalizedata.Rd @@ -63,3 +63,14 @@ The DISCBIO-class object input with the ndata and fdata slots filled. This function allows filtering of genes and cells to be used in the downstream analysis. } +\examples{ +sc <- DISCBIO(valuesG1msTest) # changes signature of data + +# In this case this function is used to normalize the reads +sc_normal <- Normalizedata( + sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, + dsn=1, rseed=17000 +) +summary(sc_normal@fdata) + +} diff --git a/man/PCAplotSymbols.Rd b/man/PCAplotSymbols.Rd index f83a6e4..531fe9c 100644 --- a/man/PCAplotSymbols.Rd +++ b/man/PCAplotSymbols.Rd @@ -21,3 +21,15 @@ Plot of the Principal Components \description{ Generates a plot of grouped PCA components } +\examples{ +sc <- DISCBIO(valuesG1msTest) +sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) +sc <- Normalizedata( + sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, + dsn=1, rseed=17000 +) +sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) +sc <- Exprmclust(sc, K=2) +PCAplotSymbols(sc) + +} diff --git a/man/comptSNE.Rd b/man/comptSNE.Rd index e79add1..d603f92 100644 --- a/man/comptSNE.Rd +++ b/man/comptSNE.Rd @@ -44,3 +44,10 @@ The DISCBIO-class object input with the tsne slot filled. This function is used to compute the t-Distributed Stochastic Neighbor Embedding (t-SNE). } +\examples{ +sc <- DISCBIO(valuesG1msTest) # changes signature of data +sc <- Clustexp(sc, cln=2) # data must be clustered before plottin +sc <- comptSNE(sc, max_iter=30) +head(sc@tsne) + +} From d560e7119307377f27e05ee5ad029ce1ed81d3a8 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Tue, 25 Aug 2020 10:45:56 +0200 Subject: [PATCH 95/99] Removed examples that need larger datasets --- R/DIscBIO-generic-Exprmclust.R | 10 ---------- R/DIscBIO-generic-PCAplotSymbols.R | 10 ---------- man/PCAplotSymbols.Rd | 12 ------------ 3 files changed, 32 deletions(-) diff --git a/R/DIscBIO-generic-Exprmclust.R b/R/DIscBIO-generic-Exprmclust.R index e28a435..0530792 100644 --- a/R/DIscBIO-generic-Exprmclust.R +++ b/R/DIscBIO-generic-Exprmclust.R @@ -18,16 +18,6 @@ #' the MBclusters slot filled. If the `object` is a data frame, the function #' returns a named list containing the four objects that together correspond #' to the contents of the MBclusters slot. -#' @examples -#' sc <- DISCBIO(valuesG1msTest) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE, plot=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- Exprmclust(sc, reduce=FALSE, K=1) -#' print(sc@MBclusters) #' setGeneric( name = "Exprmclust", diff --git a/R/DIscBIO-generic-PCAplotSymbols.R b/R/DIscBIO-generic-PCAplotSymbols.R index 52b068c..0de0caf 100644 --- a/R/DIscBIO-generic-PCAplotSymbols.R +++ b/R/DIscBIO-generic-PCAplotSymbols.R @@ -6,16 +6,6 @@ #' @importFrom grDevices rainbow #' @importFrom graphics legend #' @return Plot of the Principal Components -#' @examples -#' sc <- DISCBIO(valuesG1msRed) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- Exprmclust(sc, K=2) -#' PCAplotSymbols(sc) #' setGeneric("PCAplotSymbols", function(object, types = NULL) standardGeneric("PCAplotSymbols")) diff --git a/man/PCAplotSymbols.Rd b/man/PCAplotSymbols.Rd index 531fe9c..f83a6e4 100644 --- a/man/PCAplotSymbols.Rd +++ b/man/PCAplotSymbols.Rd @@ -21,15 +21,3 @@ Plot of the Principal Components \description{ Generates a plot of grouped PCA components } -\examples{ -sc <- DISCBIO(valuesG1msTest) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -sc <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, - dsn=1, rseed=17000 -) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -sc <- Exprmclust(sc, K=2) -PCAplotSymbols(sc) - -} From c9add4ed62192a15efcb32ff6e38cdd5e053dc86 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Thu, 27 Aug 2020 11:05:52 +0200 Subject: [PATCH 96/99] Added CRAN badge --- README.md | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/README.md b/README.md index b631030..97f3471 100644 --- a/README.md +++ b/README.md @@ -1,4 +1,4 @@ -[![Binder](https://mybinder.org/badge_logo.svg)](https://mybinder.org/v2/gh/ocbe-uio/DIscBIO/dev?filepath=notebook) [![Build Status](https://travis-ci.org/ocbe-uio/DIscBIO.svg?branch=dev)](https://travis-ci.org/ocbe-uio/DIscBIO) +![Current CRAN release](https://www.r-pkg.org/badges/version/DIscBIO) [![Binder](https://mybinder.org/badge_logo.svg)](https://mybinder.org/v2/gh/ocbe-uio/DIscBIO/dev?filepath=notebook) [![Build Status](https://travis-ci.org/ocbe-uio/DIscBIO.svg?branch=dev)](https://travis-ci.org/ocbe-uio/DIscBIO) # DIscBIO @@ -54,7 +54,7 @@ A step-by-step tutorial of DIscBIO is under construction as a standalone R vigne In order to use the Binder version of DIscBIO, just click on the badge below: -[![Binder](https://mybinder.org/badge_logo.svg)](https://mybinder.org/v2/gh/ocbe-uio/DIscBIO/dev?filepath=notebook) +[![Binder](https://mybinder.org/badge_logo.svg)](https://mybinder.org/v2/gh/ocbe-uio/DIscBIO/dev?filepath=notebook) # Development From 6732ed9721c0b8f8a4d17caa45c85253d8c94b3e Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Thu, 27 Aug 2020 11:28:01 +0200 Subject: [PATCH 97/99] Added BiocManager to Suggests (for Travis) --- DESCRIPTION | 1 + 1 file changed, 1 insertion(+) diff --git a/DESCRIPTION b/DESCRIPTION index d957fc6..c6140ad 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -45,6 +45,7 @@ Depends: R (>= 4.0), SingleCellExperiment Suggests: testthat, + BiocManager, Seurat LazyData: true RoxygenNote: 7.1.1 From c5c8578fccdeb5a05da1c01237cc3adfec2f54ea Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Thu, 27 Aug 2020 11:44:02 +0200 Subject: [PATCH 98/99] Removed BiocManager again (it didn't fix Travis after all) --- DESCRIPTION | 1 - 1 file changed, 1 deletion(-) diff --git a/DESCRIPTION b/DESCRIPTION index c6140ad..d957fc6 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -45,7 +45,6 @@ Depends: R (>= 4.0), SingleCellExperiment Suggests: testthat, - BiocManager, Seurat LazyData: true RoxygenNote: 7.1.1 From 4945b4c4641dc3a5c30c770edd71df2daaacd77d Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Thu, 27 Aug 2020 11:51:32 +0200 Subject: [PATCH 99/99] Added bioc usage to Travis checks --- .travis.yml | 4 +++- 1 file changed, 3 insertions(+), 1 deletion(-) diff --git a/.travis.yml b/.travis.yml index 5a35725..fb80073 100644 --- a/.travis.yml +++ b/.travis.yml @@ -6,4 +6,6 @@ cache: timeout: 3000 r: - release - - devel \ No newline at end of file + - devel +bioc_required: true +use_bioc: true \ No newline at end of file

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z;*^n$i!Lr-zZq_ASBUVoW{z~)$Ej4kj9X}paY*{O+U;8AFGL=Z7<^igc>Exk=sXa`MYaE3DwpX1d_P;1zEV!voER7)bCINUHRlCwI5pa_?N}PiycsCmy^~2-|J(_MwRj<*{8ysABY`nr(lXcP|`CYG-BSeQefTvt-=6fw5C~~XHU=7hxGa8n({h5U;;Mk7-W5Hu3!N*-2-`TjJ!~_7Q)i)CMZi(N#hpW^D67SCg-BPPQQ{Fr#sOS{;J!Pt2mRkkMTnDEb8suGaJBWd857R z)L}W`2vb2((auT5X)cM5ei(>;FX4ZHSu7vQJS(!%*>-U6P0Uxd_Jez2U?C#1!S!3< zkpbCpaXP@%&|VaF{;l2)D!e_oCodPuJOx^U)&Dl(0tF!CFMvPtU&tZ=*z~XDkHY_O z4dVRL`xov%$bWnl#e8k>)?Um?fb6jIy#};FLXHQReE(KKiob*pz?PXmeYk6RDwG-X z8x{D6#eebo#rr=!X0E&fUJO4-%kMa0;qT5|Cp%%vb^>i;g13c{R{IS$^VD`gkA8^$I4hWO*~my3Tn zhW)|*4x~_86)TATPae8*PeMrgBz(TDRGfYf;ez9OeE)vF$6)82xf~tWEy3zcOuNwA zx3efYxqz78)ex`{5qZHbF9!si6?xGLN?-q${3_aie*=qOCa3<<`%lsSFTj68{xbXx z`5#pf7<3R&6w?mgBWEsW$MtK!ixVKH_=J83ke)iA?Hreo&>!}1Q_*jm|Hb&f=~+4b zuWtT9R{!1U6PwJmetBp>u6QiB*LQ1L-MxgoR<5`cmnxh-H=k{{;=GaH@kTqpWPDZP O;DfPZ!I4A9{`!9kE!Q{z diff --git a/man/prepExampleDataset.Rd b/man/prepExampleDataset.Rd index 71d3b61..5b990b3 100644 --- a/man/prepExampleDataset.Rd +++ b/man/prepExampleDataset.Rd @@ -4,10 +4,12 @@ \alias{prepExampleDataset} \title{Prepare Example Dataset} \usage{ -prepExampleDataset(clustering) +prepExampleDataset(dataset, save = TRUE) } \arguments{ -\item{clustering}{determines the clustering method (either "K" or "MB").} +\item{dataset}{Dataset used for transformation} + +\item{save}{save results?} } \value{ rda file @@ -16,3 +18,8 @@ rda file Internal function that prepares a pre-treated dataset for use in several examples } +\examples{ +\donttest{ + DIscBIO:::prepExampleDataset(valuesG1msReduced, save=FALSE) +} +} diff --git a/man/valuesG1msReduced_treated_K.Rd b/man/valuesG1msReduced_treated_K.Rd new file mode 100644 index 0000000..a6c744a --- /dev/null +++ b/man/valuesG1msReduced_treated_K.Rd @@ -0,0 +1,9 @@ +% Generated by roxygen2: do not edit by hand +% Please edit documentation in R/datasets.R +\docType{data} +\name{valuesG1msReduced_treated_K} +\alias{valuesG1msReduced_treated_K} +\title{Single-cells data from a myxoid liposarcoma cell line (K variant)} +\description{ +Equivalent to valuesG1msReduced after treatment and K-means clustering. +} diff --git a/man/valuesG1msReduced_treated_MB.Rd b/man/valuesG1msReduced_treated_MB.Rd new file mode 100644 index 0000000..2d28a20 --- /dev/null +++ b/man/valuesG1msReduced_treated_MB.Rd @@ -0,0 +1,9 @@ +% Generated by roxygen2: do not edit by hand +% Please edit documentation in R/datasets.R +\docType{data} +\name{valuesG1msReduced_treated_MB} +\alias{valuesG1msReduced_treated_MB} +\title{Single-cells data from a myxoid liposarcoma cell line (MB variant)} +\description{ +Equivalent to valuesG1msReduced after treatment and model-based clustering. +} From 0717d9aeec634e1d974dd2ae576b1133ad89d5cd Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Thu, 2 Jul 2020 10:06:38 +0200 Subject: [PATCH 28/99] Simplified donttest examples --- R/DIscBIO-generic-ClassVectoringDT.R | 11 ++--------- R/DIscBIO-generic-MBClustDiffGenes.R | 12 ++---------- R/DIscBIO-generic-MBclustheatmap.R | 12 ++---------- R/DIscBIO-generic-plotexptsneMB.R | 12 ++---------- R/MB_Order.R | 12 ++---------- R/NetAnalysis.R | 11 ++--------- R/Networking.R | 11 ++--------- R/PPI.R | 11 ++--------- R/VolcanoPlot.R | 11 ++--------- man/ClassVectoringDT.Rd | 11 ++--------- man/MBClustDiffGenes.Rd | 12 ++---------- man/MB_Order.Rd | 12 ++---------- man/MBclustheatmap.Rd | 12 ++---------- man/NetAnalysis.Rd | 11 ++--------- man/Networking.Rd | 11 ++--------- man/PPI.Rd | 11 ++--------- man/VolcanoPlot.Rd | 11 ++--------- man/plotexptsneMB.Rd | 12 ++---------- 18 files changed, 36 insertions(+), 170 deletions(-) diff --git a/R/DIscBIO-generic-ClassVectoringDT.R b/R/DIscBIO-generic-ClassVectoringDT.R index fdc1e4f..147f4d2 100644 --- a/R/DIscBIO-generic-ClassVectoringDT.R +++ b/R/DIscBIO-generic-ClassVectoringDT.R @@ -15,15 +15,8 @@ #' @importFrom biomaRt useDataset useMart getBM #' @return A data frame. #' @examples -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- Clustexp(sc, cln=2) # K-means clustering -#' sc <- comptSNE(sc, max_iter=100) +#' data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details +#' sc <- valuesG1msReduced_treated_K #' cdiff <- DEGanalysis2clust( #' sc, Clustering="K-means", K=2, fdr=.2, name="Name", First="CL1", #' Second="CL2", export=FALSE diff --git a/R/DIscBIO-generic-MBClustDiffGenes.R b/R/DIscBIO-generic-MBClustDiffGenes.R index 317bd9c..02f7ac1 100644 --- a/R/DIscBIO-generic-MBClustDiffGenes.R +++ b/R/DIscBIO-generic-MBClustDiffGenes.R @@ -14,16 +14,8 @@ #' @return A list containing two tables. #' @examples #' \donttest{ -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- Exprmclust(sc, K=3) -#' sc <- comptsneMB(sc, max_iter=100) -#' sc <- Clustexp(sc, cln=3) +#' data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset +#' sc <- valuesG1msReduced_treated_MB #' sc <- MB_Order(sc, export = FALSE) #' cdiff <- MBClustDiffGenes(sc, K=3, fdr=.1, export=FALSE) #' str(cdiff) diff --git a/R/DIscBIO-generic-MBclustheatmap.R b/R/DIscBIO-generic-MBclustheatmap.R index eee63c0..a41c005 100644 --- a/R/DIscBIO-generic-MBclustheatmap.R +++ b/R/DIscBIO-generic-MBclustheatmap.R @@ -16,16 +16,8 @@ #' cluster order. #' @examples #' \donttest{ -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- NoiseFiltering(sc, export=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- Exprmclust(sc,K = 2) -#' sc <- comptsneMB(sc, max_iter=100) -#' sc <- Clustexp(sc, cln=3) +#' data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset +#' sc <- valuesG1msReduced_treated_MB #' sc <- MB_Order(sc, export = FALSE) #' MBclustheatmap(sc, hmethod="single") #' } diff --git a/R/DIscBIO-generic-plotexptsneMB.R b/R/DIscBIO-generic-plotexptsneMB.R index fc3ca35..e37436f 100644 --- a/R/DIscBIO-generic-plotexptsneMB.R +++ b/R/DIscBIO-generic-plotexptsneMB.R @@ -12,16 +12,8 @@ #' @return t-SNE plot for one particular gene #' @examples #' \donttest{ -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- Exprmclust(sc, K=3) -#' sc <- comptsneMB(sc, max_iter=100) -#' sc <- Clustexp(sc, cln=3) +#' data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset +#' sc <- valuesG1msReduced_treated_MB #' sc <- MB_Order(sc, export = FALSE) #' g <- 'ENSG00000001460' #' plotexptsneMB(sc, g) diff --git a/R/MB_Order.R b/R/MB_Order.R index 6e7c54b..87b56aa 100644 --- a/R/MB_Order.R +++ b/R/MB_Order.R @@ -10,16 +10,8 @@ #' @return The DISCBIO-class object input with the MBordering slot filled. #' @examples #' \donttest{ -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- Exprmclust(sc, K=2) -#' sc <- comptsneMB(sc, max_iter=100) -#' sc <- Clustexp(sc, cln=3) +#' data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset +#' sc <- valuesG1msReduced_treated_MB #' sc <- MB_Order(sc, export = FALSE) #' sc@MBordering #' } diff --git a/R/NetAnalysis.R b/R/NetAnalysis.R index 3b9be6a..639a857 100644 --- a/R/NetAnalysis.R +++ b/R/NetAnalysis.R @@ -13,15 +13,8 @@ #' @return A network analysis table #' @examples #' \donttest{ -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' sc <- FinalPreprocessing(sc, export=FALSE, quiet=TRUE) -#' sc <- Clustexp(sc, cln=3, quiet=TRUE) # K-means clustering -#' sc <- comptSNE(sc, max_iter=100, quiet=TRUE) +#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +#' sc <- valuesG1msReduced_treated_K #' dff <- DEGanalysis2clust(sc, K=3, export=FALSE, quiet=TRUE, plot=FALSE) #' ppi <- PPI(dff$FinalDEGsU[, "Gene Name"]) #' NetAnalysis(ppi, export=FALSE) diff --git a/R/Networking.R b/R/Networking.R index cbbfeac..1afbb10 100644 --- a/R/Networking.R +++ b/R/Networking.R @@ -15,15 +15,8 @@ #' @return A plot of the network #' @examples #' \donttest{ -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE, plot=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' sc <- FinalPreprocessing(sc, export=FALSE, quiet=TRUE) -#' sc <- Clustexp(sc, cln=3, quiet=TRUE) #' K-means clustering -#' sc <- comptSNE(sc, max_iter=100, quiet=TRUE) +#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +#' sc <- valuesG1msReduced_treated_K #' dff <- DEGanalysis2clust(sc, K=3, export=FALSE, quiet=TRUE, plot=FALSE) #' Networking(dff$FinalDEGsU[, "Gene Name"]) #' } diff --git a/R/PPI.R b/R/PPI.R index f61b61f..c0d8d97 100644 --- a/R/PPI.R +++ b/R/PPI.R @@ -13,15 +13,8 @@ #' information. #' @examples #' \donttest{ -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' sc <- FinalPreprocessing(sc, export=FALSE, quiet=TRUE) -#' sc <- Clustexp(sc, cln=3, quiet=TRUE) # K-means clustering -#' sc <- comptSNE(sc, max_iter=100, quiet=TRUE) +#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +#' sc <- valuesG1msReduced_treated_K #' dff <- DEGanalysis2clust(sc, K=3, export=FALSE, quiet=TRUE, plot=FALSE) #' PPI(dff$FinalDEGsU[, "Gene Name"]) #' } diff --git a/R/VolcanoPlot.R b/R/VolcanoPlot.R index 4b73f58..9cb4549 100644 --- a/R/VolcanoPlot.R +++ b/R/VolcanoPlot.R @@ -18,15 +18,8 @@ #' @export #' @examples #' \donttest{ -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE, plot=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' sc <- FinalPreprocessing(sc, export=FALSE, quiet=TRUE) -#' sc <- Clustexp(sc, cln=3, quiet=TRUE) # K-means clustering -#' sc <- comptSNE(sc, max_iter=100, quiet=TRUE) +#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +#' sc <- valuesG1msReduced_treated_K #' dff <- DEGanalysis2clust(sc, K=3, export=FALSE, quiet=TRUE, plot=FALSE) #' VolcanoPlot(dff$FinalDEGsU, value=0.05, FS=.4) #' } diff --git a/man/ClassVectoringDT.Rd b/man/ClassVectoringDT.Rd index c713447..013ff44 100644 --- a/man/ClassVectoringDT.Rd +++ b/man/ClassVectoringDT.Rd @@ -52,15 +52,8 @@ This function generates a class vector for the input dataset so the decision tree analysis can be implemented afterwards. } \examples{ -sc <- DISCBIO(valuesG1msReduced) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -sc <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, - dsn=1, rseed=17000 -) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -sc <- Clustexp(sc, cln=2) # K-means clustering -sc <- comptSNE(sc, max_iter=100) +data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details +sc <- valuesG1msReduced_treated_K cdiff <- DEGanalysis2clust( sc, Clustering="K-means", K=2, fdr=.2, name="Name", First="CL1", Second="CL2", export=FALSE diff --git a/man/MBClustDiffGenes.Rd b/man/MBClustDiffGenes.Rd index 910f567..c0a002d 100644 --- a/man/MBClustDiffGenes.Rd +++ b/man/MBClustDiffGenes.Rd @@ -45,16 +45,8 @@ ClustDiffGenes } \examples{ \donttest{ -sc <- DISCBIO(valuesG1msReduced) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -sc <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, - dsn=1, rseed=17000 -) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -sc <- Exprmclust(sc, K=3) -sc <- comptsneMB(sc, max_iter=100) -sc <- Clustexp(sc, cln=3) +data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset +sc <- valuesG1msReduced_treated_MB sc <- MB_Order(sc, export = FALSE) cdiff <- MBClustDiffGenes(sc, K=3, fdr=.1, export=FALSE) str(cdiff) diff --git a/man/MB_Order.Rd b/man/MB_Order.Rd index 5271ddb..35adaca 100644 --- a/man/MB_Order.Rd +++ b/man/MB_Order.Rd @@ -23,16 +23,8 @@ This function takes the exact output of exprmclust function and } \examples{ \donttest{ -sc <- DISCBIO(valuesG1msReduced) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -sc <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, - dsn=1, rseed=17000 -) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -sc <- Exprmclust(sc, K=2) -sc <- comptsneMB(sc, max_iter=100) -sc <- Clustexp(sc, cln=3) +data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset +sc <- valuesG1msReduced_treated_MB sc <- MB_Order(sc, export = FALSE) sc@MBordering } diff --git a/man/MBclustheatmap.Rd b/man/MBclustheatmap.Rd index 6c7bcf9..4cd5858 100644 --- a/man/MBclustheatmap.Rd +++ b/man/MBclustheatmap.Rd @@ -32,16 +32,8 @@ This functions plots a heatmap of the distance matrix grouped } \examples{ \donttest{ -sc <- DISCBIO(valuesG1msReduced) -sc <- NoiseFiltering(sc, export=FALSE) -sc <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, - dsn=1, rseed=17000 -) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -sc <- Exprmclust(sc,K = 2) -sc <- comptsneMB(sc, max_iter=100) -sc <- Clustexp(sc, cln=3) +data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset +sc <- valuesG1msReduced_treated_MB sc <- MB_Order(sc, export = FALSE) MBclustheatmap(sc, hmethod="single") } diff --git a/man/NetAnalysis.Rd b/man/NetAnalysis.Rd index 109018b..d26a2b4 100644 --- a/man/NetAnalysis.Rd +++ b/man/NetAnalysis.Rd @@ -24,15 +24,8 @@ This function checks the connectivity degree and the betweenness } \examples{ \donttest{ -sc <- DISCBIO(valuesG1msReduced) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -sc <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, - dsn=1, rseed=17000 -) -sc <- FinalPreprocessing(sc, export=FALSE, quiet=TRUE) -sc <- Clustexp(sc, cln=3, quiet=TRUE) # K-means clustering -sc <- comptSNE(sc, max_iter=100, quiet=TRUE) +data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +sc <- valuesG1msReduced_treated_K dff <- DEGanalysis2clust(sc, K=3, export=FALSE, quiet=TRUE, plot=FALSE) ppi <- PPI(dff$FinalDEGsU[, "Gene Name"]) NetAnalysis(ppi, export=FALSE) diff --git a/man/Networking.Rd b/man/Networking.Rd index 6915731..a12694a 100644 --- a/man/Networking.Rd +++ b/man/Networking.Rd @@ -33,15 +33,8 @@ This function uses STRING-api to plot the network. } \examples{ \donttest{ -sc <- DISCBIO(valuesG1msReduced) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE, plot=FALSE) -sc <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, - dsn=1, rseed=17000 -) -sc <- FinalPreprocessing(sc, export=FALSE, quiet=TRUE) -sc <- Clustexp(sc, cln=3, quiet=TRUE) #' K-means clustering -sc <- comptSNE(sc, max_iter=100, quiet=TRUE) +data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +sc <- valuesG1msReduced_treated_K dff <- DEGanalysis2clust(sc, K=3, export=FALSE, quiet=TRUE, plot=FALSE) Networking(dff$FinalDEGsU[, "Gene Name"]) } diff --git a/man/PPI.Rd b/man/PPI.Rd index 9646c95..7b163d2 100644 --- a/man/PPI.Rd +++ b/man/PPI.Rd @@ -25,15 +25,8 @@ This function uses STRING-api. The outcome of STRING analysis } \examples{ \donttest{ -sc <- DISCBIO(valuesG1msReduced) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -sc <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, - dsn=1, rseed=17000 -) -sc <- FinalPreprocessing(sc, export=FALSE, quiet=TRUE) -sc <- Clustexp(sc, cln=3, quiet=TRUE) # K-means clustering -sc <- comptSNE(sc, max_iter=100, quiet=TRUE) +data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +sc <- valuesG1msReduced_treated_K dff <- DEGanalysis2clust(sc, K=3, export=FALSE, quiet=TRUE, plot=FALSE) PPI(dff$FinalDEGsU[, "Gene Name"]) } diff --git a/man/VolcanoPlot.Rd b/man/VolcanoPlot.Rd index f733247..37387ee 100644 --- a/man/VolcanoPlot.Rd +++ b/man/VolcanoPlot.Rd @@ -29,15 +29,8 @@ Plotting differentially expressed genes (DEGs) in a particular } \examples{ \donttest{ -sc <- DISCBIO(valuesG1msReduced) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE, plot=FALSE) -sc <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, - dsn=1, rseed=17000 -) -sc <- FinalPreprocessing(sc, export=FALSE, quiet=TRUE) -sc <- Clustexp(sc, cln=3, quiet=TRUE) # K-means clustering -sc <- comptSNE(sc, max_iter=100, quiet=TRUE) +data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +sc <- valuesG1msReduced_treated_K dff <- DEGanalysis2clust(sc, K=3, export=FALSE, quiet=TRUE, plot=FALSE) VolcanoPlot(dff$FinalDEGsU, value=0.05, FS=.4) } diff --git a/man/plotexptsneMB.Rd b/man/plotexptsneMB.Rd index e71ed2c..ca78123 100644 --- a/man/plotexptsneMB.Rd +++ b/man/plotexptsneMB.Rd @@ -30,16 +30,8 @@ The t-SNE map representation can also be used to analyze } \examples{ \donttest{ -sc <- DISCBIO(valuesG1msReduced) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -sc <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, - dsn=1, rseed=17000 -) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -sc <- Exprmclust(sc, K=3) -sc <- comptsneMB(sc, max_iter=100) -sc <- Clustexp(sc, cln=3) +data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset +sc <- valuesG1msReduced_treated_MB sc <- MB_Order(sc, export = FALSE) g <- 'ENSG00000001460' plotexptsneMB(sc, g) From 0963cb42f157c31e7353b40b4aa46caf83ec1a65 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Thu, 2 Jul 2020 10:20:03 +0200 Subject: [PATCH 29/99] Replaced dataset in other examples --- R/DIscBIO-generic-DEGanalysis.R | 11 ++--------- R/DIscBIO-generic-DEGanalysis2clust.R | 11 ++--------- R/DIscBIO-generic-KMclustheatmap.R | 5 ++--- R/DIscBIO-generic-KmeanOrder.R | 4 ++-- R/DIscBIO-generic-PlotmclustMB.R | 10 ++-------- R/DIscBIO-generic-plotExptSNE.R | 5 ++--- R/DIscBIO-generic-plotGap.R | 4 ++-- R/DIscBIO-generic-plotKmeansLabelstSNE.R | 5 ++--- R/DIscBIO-generic-plotMBLabelstSNE.R | 11 ++--------- R/DIscBIO-generic-plotOrderKMtsne.R | 5 ++--- R/DIscBIO-generic-plotOrderMBtsne.R | 8 ++------ R/DIscBIO-generic-plotSilhouette.R | 5 ++--- R/DIscBIO-generic-plotSymbolstSNE.R | 5 ++--- R/DIscBIO-generic-plotsilhouetteMB.R | 10 ++-------- R/DIscBIO-generic-plottSNE.R | 5 ++--- R/DIscBIO-generic-plottsneMB.R | 11 ++--------- R/J48DT.R | 11 ++--------- R/J48DTeval.R | 11 ++--------- R/Jaccard.R | 4 ++-- R/PlotMBexpPCA.R | 7 ++----- R/PlotMBorderPCA.R | 11 ++--------- R/RpartDT.R | 11 ++--------- R/RpartEVAL.R | 11 ++--------- man/DEGanalysis.Rd | 11 ++--------- man/DEGanalysis2clust.Rd | 11 ++--------- man/J48DT.Rd | 11 ++--------- man/J48DTeval.Rd | 11 ++--------- man/Jaccard.Rd | 4 ++-- man/KMclustheatmap.Rd | 5 ++--- man/KmeanOrder.Rd | 4 ++-- man/PlotMBexpPCA.Rd | 7 ++----- man/PlotMBorderPCA.Rd | 11 ++--------- man/PlotmclustMB.Rd | 10 ++-------- man/RpartDT.Rd | 11 ++--------- man/RpartEVAL.Rd | 11 ++--------- man/plotExptSNE.Rd | 5 ++--- man/plotGap.Rd | 4 ++-- man/plotKmeansLabelstSNE.Rd | 5 ++--- man/plotMBLabelstSNE.Rd | 11 ++--------- man/plotOrderKMtsne.Rd | 5 ++--- man/plotOrderMBtsne.Rd | 8 ++------ man/plotSilhouette.Rd | 5 ++--- man/plotSymbolstSNE.Rd | 5 ++--- man/plotsilhouetteMB.Rd | 10 ++-------- man/plottSNE.Rd | 5 ++--- man/plottsneMB.Rd | 11 ++--------- 46 files changed, 92 insertions(+), 270 deletions(-) diff --git a/R/DIscBIO-generic-DEGanalysis.R b/R/DIscBIO-generic-DEGanalysis.R index 4364e25..57384a1 100644 --- a/R/DIscBIO-generic-DEGanalysis.R +++ b/R/DIscBIO-generic-DEGanalysis.R @@ -20,15 +20,8 @@ #' @param ... additional parameters to be passed to samr() #' @return A list containing two tables. #' @examples -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' sc <- Clustexp(sc, cln=3) # K-means clustering -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- comptSNE(sc, max_iter=100) +#' data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details +#' sc <- valuesG1msReduced_treated_K #' DEGanalysis( #' sc, Clustering="K-means", K=3, fdr=0.1, name="Name", export = FALSE, #' nresamp=5, nperms=20 diff --git a/R/DIscBIO-generic-DEGanalysis2clust.R b/R/DIscBIO-generic-DEGanalysis2clust.R index bb1bb77..536b7d7 100644 --- a/R/DIscBIO-generic-DEGanalysis2clust.R +++ b/R/DIscBIO-generic-DEGanalysis2clust.R @@ -25,15 +25,8 @@ #' @importFrom AnnotationDbi keys #' @return A list containing two tables. #' @examples -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- Clustexp(sc, cln=3) # K-means clustering -#' sc <- comptSNE(sc, max_iter=100) +#' data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details +#' sc <- valuesG1msReduced_treated_K #' DEGanalysis2clust( #' sc, Clustering="K-means", K=3, fdr=0.1, name="Name", export = FALSE #' ) diff --git a/R/DIscBIO-generic-KMclustheatmap.R b/R/DIscBIO-generic-KMclustheatmap.R index 3c1fd57..7ee5196 100644 --- a/R/DIscBIO-generic-KMclustheatmap.R +++ b/R/DIscBIO-generic-KMclustheatmap.R @@ -12,9 +12,8 @@ #' cluster order. #' @importFrom stats hclust as.dist cor #' @examples -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- Clustexp(sc, cln=3, quiet=TRUE) # K-means clustering -#' sc <- comptSNE(sc, max_iter=100, quiet=TRUE) +#' data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details +#' sc <- valuesG1msReduced_treated_K #' KMclustheatmap(sc, hmethod="single") setGeneric( "KMclustheatmap", diff --git a/R/DIscBIO-generic-KmeanOrder.R b/R/DIscBIO-generic-KmeanOrder.R index 140e2d0..96ee938 100644 --- a/R/DIscBIO-generic-KmeanOrder.R +++ b/R/DIscBIO-generic-KmeanOrder.R @@ -8,8 +8,8 @@ #' @importFrom TSCAN TSCANorder #' @return The DISCBIO-class object input with the kordering slot filled. #' @examples -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- Clustexp(sc, cln=3) # K-means clustering +#' data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details +#' sc <- valuesG1msReduced_treated_K #' Order <- KmeanOrder(sc, export = FALSE) #' Order@kordering setGeneric("KmeanOrder", function(object, quiet = FALSE, export = TRUE) diff --git a/R/DIscBIO-generic-PlotmclustMB.R b/R/DIscBIO-generic-PlotmclustMB.R index 7a640b9..a7d5eda 100644 --- a/R/DIscBIO-generic-PlotmclustMB.R +++ b/R/DIscBIO-generic-PlotmclustMB.R @@ -7,14 +7,8 @@ #' @importFrom igraph get.edgelist degree get.shortest.paths #' @return A plot of the PCA. #' @examples -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- Exprmclust(sc, K=2) +#' data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset +#' sc <- valuesG1msReduced_treated_MB #' PlotmclustMB(sc) setGeneric("PlotmclustMB", function(object) standardGeneric("PlotmclustMB")) diff --git a/R/DIscBIO-generic-plotExptSNE.R b/R/DIscBIO-generic-plotExptSNE.R index b73acae..1f8e6f5 100644 --- a/R/DIscBIO-generic-plotExptSNE.R +++ b/R/DIscBIO-generic-plotExptSNE.R @@ -10,9 +10,8 @@ #' NULL and the first element of \code{g} is chosen. #' @return t-SNE plot for one particular gene #' @examples -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- Clustexp(sc, cln=3, quiet=TRUE) # K-means clustering -#' sc <- comptSNE(sc, max_iter=100) +#' data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details +#' sc <- valuesG1msReduced_treated_K #' g <- 'ENSG00000001460' #' plotExptSNE(sc, g) diff --git a/R/DIscBIO-generic-plotGap.R b/R/DIscBIO-generic-plotGap.R index 91e2c69..70ad424 100644 --- a/R/DIscBIO-generic-plotGap.R +++ b/R/DIscBIO-generic-plotGap.R @@ -5,8 +5,8 @@ #' @param y_limits 2-length numeric vector with the limits for the gap plot #' @return A plot of the gap statistics #' @examples -#' sc <- DISCBIO(valuesG1msReduced) # changes signature of data -#' sc <- Clustexp(sc, cln=3) # data must be clustered before plotting +#' data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details +#' sc <- valuesG1msReduced_treated_K #' plotGap(sc) setGeneric( "plotGap", function(object, y_limits = NULL) standardGeneric("plotGap") diff --git a/R/DIscBIO-generic-plotKmeansLabelstSNE.R b/R/DIscBIO-generic-plotKmeansLabelstSNE.R index be29590..47d7fc6 100644 --- a/R/DIscBIO-generic-plotKmeansLabelstSNE.R +++ b/R/DIscBIO-generic-plotKmeansLabelstSNE.R @@ -5,9 +5,8 @@ #' @importFrom graphics text #' @return Plot containing the ID of the cells in each cluster #' @examples -#' sc <- DISCBIO(valuesG1msReduced) # changes signature of data -#' sc <- Clustexp(sc, cln=3) # data must be clustered before plottin -#' sc <- comptSNE(sc, max_iter=100) +#' data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details +#' sc <- valuesG1msReduced_treated_K #' plotKmeansLabelstSNE(sc) # Plots the ID of the cells in each cluster setGeneric("plotKmeansLabelstSNE", function(object) standardGeneric("plotKmeansLabelstSNE")) diff --git a/R/DIscBIO-generic-plotMBLabelstSNE.R b/R/DIscBIO-generic-plotMBLabelstSNE.R index 2cdcbbd..838349b 100644 --- a/R/DIscBIO-generic-plotMBLabelstSNE.R +++ b/R/DIscBIO-generic-plotMBLabelstSNE.R @@ -4,15 +4,8 @@ #' @importFrom graphics text #' @return A plot of the `object@MBtsne` values #' @examples -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- Exprmclust(sc, K=2) -#' sc <- comptsneMB(sc, rseed=15555, quiet=TRUE, max_iter=100) +#' data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset +#' sc <- valuesG1msReduced_treated_MB #' plotMBLabelstSNE(sc) setGeneric("plotMBLabelstSNE", function(object) standardGeneric("plotMBLabelstSNE") diff --git a/R/DIscBIO-generic-plotOrderKMtsne.R b/R/DIscBIO-generic-plotOrderKMtsne.R index f5319ca..ce5e946 100644 --- a/R/DIscBIO-generic-plotOrderKMtsne.R +++ b/R/DIscBIO-generic-plotOrderKMtsne.R @@ -5,9 +5,8 @@ #' @param object \code{DISCBIO} class object. #' @return A plot of the pseudo-time ordering. #' @examples -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- Clustexp(sc, cln=3) # K-means clustering -#' sc <- comptSNE(sc, max_iter=100) +#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +#' sc <- valuesG1msReduced_treated_K #' sc <- KmeanOrder(sc, export = FALSE) #' plotOrderKMtsne(sc) setGeneric("plotOrderKMtsne", function(object) diff --git a/R/DIscBIO-generic-plotOrderMBtsne.R b/R/DIscBIO-generic-plotOrderMBtsne.R index ea7d226..26d4388 100644 --- a/R/DIscBIO-generic-plotOrderMBtsne.R +++ b/R/DIscBIO-generic-plotOrderMBtsne.R @@ -5,12 +5,8 @@ #' @param object \code{DISCBIO} class object. #' @return A plot of the pseudo-time ordering. #' @examples -#' sc<- DISCBIO(valuesG1msReduced) -#' sc<-NoiseFiltering(sc,percentile=0.9, CV=0.2, export=FALSE) -#' sc<-Normalizedata(sc) -#' sc<-FinalPreprocessing(sc,GeneFlitering="NoiseF", export=FALSE) -#' sc <- Exprmclust(sc, K=2, reduce=TRUE, quiet=TRUE) -#' sc<- comptsneMB(sc, rseed=15555, quiet=TRUE, max_iter=100) +#' data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset +#' sc <- valuesG1msReduced_treated_MB #' plotOrderMBtsne(sc) setGeneric("plotOrderMBtsne", function(object) standardGeneric("plotOrderMBtsne")) diff --git a/R/DIscBIO-generic-plotSilhouette.R b/R/DIscBIO-generic-plotSilhouette.R index 58f5457..c1249ea 100644 --- a/R/DIscBIO-generic-plotSilhouette.R +++ b/R/DIscBIO-generic-plotSilhouette.R @@ -12,9 +12,8 @@ #' @importFrom cluster silhouette #' @return A silhouette plot #' @examples -#' sc <- DISCBIO(valuesG1msReduced) # changes signature of data -#' sc <- Clustexp(sc, cln=3) # data must be clustered before plottin -#' sc <- comptSNE(sc, max_iter=100) +#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +#' sc <- valuesG1msReduced_treated_K #' plotSilhouette(sc, K=3) setGeneric( name = "plotSilhouette", diff --git a/R/DIscBIO-generic-plotSymbolstSNE.R b/R/DIscBIO-generic-plotSymbolstSNE.R index 0ab56f7..adefae8 100644 --- a/R/DIscBIO-generic-plotSymbolstSNE.R +++ b/R/DIscBIO-generic-plotSymbolstSNE.R @@ -7,9 +7,8 @@ #' "left", "topleft", "top", "topright", "right" and "center". Default is #' "bottomright" #' @examples -#' sc <- DISCBIO(valuesG1msReduced) # changes signature of data -#' sc <- Clustexp(sc, cln=3) # data must be clustered before plottin -#' sc <- comptSNE(sc, max_iter=100, quiet=TRUE) +#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +#' sc <- valuesG1msReduced_treated_K #' plotSymbolstSNE(sc,types=sub("(\\_\\d+)$","", names(sc@ndata))) setGeneric( "plotSymbolstSNE", diff --git a/R/DIscBIO-generic-plotsilhouetteMB.R b/R/DIscBIO-generic-plotsilhouetteMB.R index de3338e..52ad55f 100644 --- a/R/DIscBIO-generic-plotsilhouetteMB.R +++ b/R/DIscBIO-generic-plotsilhouetteMB.R @@ -12,14 +12,8 @@ #' @importFrom cluster silhouette #' @return A silhouette plot #' @examples -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, -#' downsample=FALSE, dsn=1, rseed=17000 -#' ) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- Exprmclust(sc, K=2, reduce=TRUE, quiet=TRUE) +#' data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset +#' sc <- valuesG1msReduced_treated_MB #' plotsilhouetteMB(sc, K=2) setGeneric("plotsilhouetteMB", function(object, K) standardGeneric("plotsilhouetteMB")) diff --git a/R/DIscBIO-generic-plottSNE.R b/R/DIscBIO-generic-plottSNE.R index 3196498..15414d9 100644 --- a/R/DIscBIO-generic-plottSNE.R +++ b/R/DIscBIO-generic-plottSNE.R @@ -4,9 +4,8 @@ #' @importFrom graphics text #' @return A plot of t-SNEs. #' @examples -#' sc <- DISCBIO(valuesG1msReduced) # changes signature of data -#' sc <- Clustexp(sc, cln=3) # data must be clustered before plotting -#' sc <- comptSNE(sc, max_iter=100, quiet=TRUE) +#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +#' sc <- valuesG1msReduced_treated_K #' plottSNE(sc) setGeneric("plottSNE", function(object) standardGeneric("plottSNE")) diff --git a/R/DIscBIO-generic-plottsneMB.R b/R/DIscBIO-generic-plottsneMB.R index e70956c..3341658 100644 --- a/R/DIscBIO-generic-plottsneMB.R +++ b/R/DIscBIO-generic-plottsneMB.R @@ -5,15 +5,8 @@ #' @importFrom graphics text #' @return A plot of t-SNEs. #' @examples -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- Exprmclust(sc, K=2) -#' sc <- comptsneMB(sc, rseed=15555, quiet=TRUE, max_iter=100) +#' data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset +#' sc <- valuesG1msReduced_treated_MB #' plottsneMB(sc) setGeneric( name = "plottsneMB", diff --git a/R/J48DT.R b/R/J48DT.R index c62015c..f120e3a 100644 --- a/R/J48DT.R +++ b/R/J48DT.R @@ -13,15 +13,8 @@ #' @return Information about the J48 model and, by default, a plot of the #' decision tree. #' @examples -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- Clustexp(sc, cln=3) # K-means clustering -#' sc <- comptSNE(sc, max_iter=100) +#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +#' sc <- valuesG1msReduced_treated_K #' cdiff <- DEGanalysis2clust( #' sc, Clustering="K-means", K=3, fdr=.2, name="Name", First="CL1", #' Second="CL2", export=FALSE, diff --git a/R/J48DTeval.R b/R/J48DTeval.R index 3ccbf70..26814f8 100644 --- a/R/J48DTeval.R +++ b/R/J48DTeval.R @@ -13,15 +13,8 @@ #' @importFrom stats predict #' @return Statistics about the J48 model #' @examples -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- Clustexp(sc, cln=3) # K-means clustering -#' sc <- comptSNE(sc, max_iter=100) +#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +#' sc <- valuesG1msReduced_treated_K #' cdiff <- DEGanalysis2clust( #' sc, Clustering="K-means", K=3, fdr=.2, name="Name", First="CL1", #' Second="CL2", export=FALSE diff --git a/R/Jaccard.R b/R/Jaccard.R index b4fd7ea..5c82279 100644 --- a/R/Jaccard.R +++ b/R/Jaccard.R @@ -16,8 +16,8 @@ #' @importFrom graphics barplot box #' @return A plot of the mean Jaccard similarity coefficient per cluster. #' @examples -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- Clustexp(sc, cln=3, quiet=TRUE) # K-means clustering +#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +#' sc <- valuesG1msReduced_treated_K #' Jaccard(sc, Clustering="K-means", K=3) Jaccard <- function( object, diff --git a/R/PlotMBexpPCA.R b/R/PlotMBexpPCA.R index 30d98f2..7909499 100644 --- a/R/PlotMBexpPCA.R +++ b/R/PlotMBexpPCA.R @@ -12,11 +12,8 @@ #' @importFrom graphics layout par image #' @return A plot of the PCA. #' @examples -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -#' sc <- Normalizedata(sc) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- Exprmclust(sc, K=2, reduce=TRUE, quiet=TRUE) +#' data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset +#' sc <- valuesG1msReduced_treated_MB #' g <- "ENSG00000010244" # Plotting the expression of MT-RNR2 #' PlotMBexpPCA(sc, g) #' @export diff --git a/R/PlotMBorderPCA.R b/R/PlotMBorderPCA.R index 3e7e103..ef91613 100644 --- a/R/PlotMBorderPCA.R +++ b/R/PlotMBorderPCA.R @@ -8,15 +8,8 @@ #' @export #' @return A plot of the PCA. #' @examples -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- Exprmclust(sc, K=3) -#' sc <- Clustexp(sc, cln=3) +#' data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset +#' sc <- valuesG1msReduced_treated_MB #' sc <- MB_Order(sc, export = FALSE) #' PlotMBorderPCA(sc) diff --git a/R/RpartDT.R b/R/RpartDT.R index 269c0aa..89256d2 100644 --- a/R/RpartDT.R +++ b/R/RpartDT.R @@ -11,15 +11,8 @@ #' @return Information about the model and, by default, a plot of the decision #' tree. #' @examples -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- Clustexp(sc, cln=3) # K-means clustering -#' sc <- comptSNE(sc, max_iter=100) +#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +#' sc <- valuesG1msReduced_treated_K #' cdiff <- DEGanalysis2clust( #' sc, Clustering="K-means", K=3, fdr=.2, name="Name", First="CL1", #' Second="CL2", export=FALSE diff --git a/R/RpartEVAL.R b/R/RpartEVAL.R index 3f10349..724405e 100644 --- a/R/RpartEVAL.R +++ b/R/RpartEVAL.R @@ -13,15 +13,8 @@ #' @importFrom stats predict #' @return Performance statistics of the model #' @examples -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- Clustexp(sc, cln=3) # K-means clustering -#' sc <- comptSNE(sc, max_iter=100) +#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +#' sc <- valuesG1msReduced_treated_K #' cdiff <- DEGanalysis2clust( #' sc, Clustering="K-means", K=3, fdr=.2, name="Name", First="CL1", #' Second="CL2", export=FALSE diff --git a/man/DEGanalysis.Rd b/man/DEGanalysis.Rd index 9339b6d..ba717ad 100644 --- a/man/DEGanalysis.Rd +++ b/man/DEGanalysis.Rd @@ -60,15 +60,8 @@ This function defines DEGs between all individual clusters generated by either K-means or model based clustering. } \examples{ -sc <- DISCBIO(valuesG1msReduced) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -sc <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, - dsn=1, rseed=17000 -) -sc <- Clustexp(sc, cln=3) # K-means clustering -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -sc <- comptSNE(sc, max_iter=100) +data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details +sc <- valuesG1msReduced_treated_K DEGanalysis( sc, Clustering="K-means", K=3, fdr=0.1, name="Name", export = FALSE, nresamp=5, nperms=20 diff --git a/man/DEGanalysis2clust.Rd b/man/DEGanalysis2clust.Rd index e9c9ce8..9a2d461 100644 --- a/man/DEGanalysis2clust.Rd +++ b/man/DEGanalysis2clust.Rd @@ -70,15 +70,8 @@ This function defines DEGs between particular clusters generated by either K-means or model based clustering. } \examples{ -sc <- DISCBIO(valuesG1msReduced) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -sc <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, - dsn=1, rseed=17000 -) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -sc <- Clustexp(sc, cln=3) # K-means clustering -sc <- comptSNE(sc, max_iter=100) +data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details +sc <- valuesG1msReduced_treated_K DEGanalysis2clust( sc, Clustering="K-means", K=3, fdr=0.1, name="Name", export = FALSE ) diff --git a/man/J48DT.Rd b/man/J48DT.Rd index 5fabfcc..c5225e1 100644 --- a/man/J48DT.Rd +++ b/man/J48DT.Rd @@ -23,15 +23,8 @@ The decision tree analysis is implemented over a training binomial differential expression. } \examples{ -sc <- DISCBIO(valuesG1msReduced) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -sc <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, - dsn=1, rseed=17000 -) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -sc <- Clustexp(sc, cln=3) # K-means clustering -sc <- comptSNE(sc, max_iter=100) +data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +sc <- valuesG1msReduced_treated_K cdiff <- DEGanalysis2clust( sc, Clustering="K-means", K=3, fdr=.2, name="Name", First="CL1", Second="CL2", export=FALSE, diff --git a/man/J48DTeval.Rd b/man/J48DTeval.Rd index 669f593..f6c648c 100644 --- a/man/J48DTeval.Rd +++ b/man/J48DTeval.Rd @@ -28,15 +28,8 @@ This function evaluates the performance of the generated trees for error estimation by ten-fold cross validation assessment. } \examples{ -sc <- DISCBIO(valuesG1msReduced) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -sc <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, - dsn=1, rseed=17000 -) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -sc <- Clustexp(sc, cln=3) # K-means clustering -sc <- comptSNE(sc, max_iter=100) +data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +sc <- valuesG1msReduced_treated_K cdiff <- DEGanalysis2clust( sc, Clustering="K-means", K=3, fdr=.2, name="Name", First="CL1", Second="CL2", export=FALSE diff --git a/man/Jaccard.Rd b/man/Jaccard.Rd index 56dc870..2d5a0cd 100644 --- a/man/Jaccard.Rd +++ b/man/Jaccard.Rd @@ -30,7 +30,7 @@ Robustness of the clusters can be assessed by Jaccard’s clusters divided by the union. } \examples{ -sc <- DISCBIO(valuesG1msReduced) -sc <- Clustexp(sc, cln=3, quiet=TRUE) # K-means clustering +data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +sc <- valuesG1msReduced_treated_K Jaccard(sc, Clustering="K-means", K=3) } diff --git a/man/KMclustheatmap.Rd b/man/KMclustheatmap.Rd index 4c3137c..01e0f56 100644 --- a/man/KMclustheatmap.Rd +++ b/man/KMclustheatmap.Rd @@ -29,8 +29,7 @@ This functions plots a heatmap of the distance matrix grouped the x and y-axes. } \examples{ -sc <- DISCBIO(valuesG1msReduced) -sc <- Clustexp(sc, cln=3, quiet=TRUE) # K-means clustering -sc <- comptSNE(sc, max_iter=100, quiet=TRUE) +data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details +sc <- valuesG1msReduced_treated_K KMclustheatmap(sc, hmethod="single") } diff --git a/man/KmeanOrder.Rd b/man/KmeanOrder.Rd index 2a687e7..2674efe 100644 --- a/man/KmeanOrder.Rd +++ b/man/KmeanOrder.Rd @@ -25,8 +25,8 @@ This function takes the exact output of exprmclust function and connects cluster centers. } \examples{ -sc <- DISCBIO(valuesG1msReduced) -sc <- Clustexp(sc, cln=3) # K-means clustering +data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details +sc <- valuesG1msReduced_treated_K Order <- KmeanOrder(sc, export = FALSE) Order@kordering } diff --git a/man/PlotMBexpPCA.Rd b/man/PlotMBexpPCA.Rd index 0839fc4..6b7e48c 100644 --- a/man/PlotMBexpPCA.Rd +++ b/man/PlotMBexpPCA.Rd @@ -24,11 +24,8 @@ The PCA representation can also be used to show the gene expression of a particular gene. } \examples{ -sc <- DISCBIO(valuesG1msReduced) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -sc <- Normalizedata(sc) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -sc <- Exprmclust(sc, K=2, reduce=TRUE, quiet=TRUE) +data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset +sc <- valuesG1msReduced_treated_MB g <- "ENSG00000010244" # Plotting the expression of MT-RNR2 PlotMBexpPCA(sc, g) } diff --git a/man/PlotMBorderPCA.Rd b/man/PlotMBorderPCA.Rd index 5b1c728..43c540a 100644 --- a/man/PlotMBorderPCA.Rd +++ b/man/PlotMBorderPCA.Rd @@ -17,15 +17,8 @@ The PCA representation can also be used to show the pseudo-time ordering. } \examples{ -sc <- DISCBIO(valuesG1msReduced) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -sc <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, - dsn=1, rseed=17000 -) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -sc <- Exprmclust(sc, K=3) -sc <- Clustexp(sc, cln=3) +data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset +sc <- valuesG1msReduced_treated_MB sc <- MB_Order(sc, export = FALSE) PlotMBorderPCA(sc) } diff --git a/man/PlotmclustMB.Rd b/man/PlotmclustMB.Rd index 6838fd5..c62a0bf 100644 --- a/man/PlotmclustMB.Rd +++ b/man/PlotmclustMB.Rd @@ -19,13 +19,7 @@ A plot of the PCA. Plot the model-based clustering results } \examples{ -sc <- DISCBIO(valuesG1msReduced) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -sc <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, - dsn=1, rseed=17000 -) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -sc <- Exprmclust(sc, K=2) +data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset +sc <- valuesG1msReduced_treated_MB PlotmclustMB(sc) } diff --git a/man/RpartDT.Rd b/man/RpartDT.Rd index 70d0426..fa97410 100644 --- a/man/RpartDT.Rd +++ b/man/RpartDT.Rd @@ -23,15 +23,8 @@ The decision tree analysis is implemented over a training binomial differential expression. } \examples{ -sc <- DISCBIO(valuesG1msReduced) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -sc <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, - dsn=1, rseed=17000 -) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -sc <- Clustexp(sc, cln=3) # K-means clustering -sc <- comptSNE(sc, max_iter=100) +data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +sc <- valuesG1msReduced_treated_K cdiff <- DEGanalysis2clust( sc, Clustering="K-means", K=3, fdr=.2, name="Name", First="CL1", Second="CL2", export=FALSE diff --git a/man/RpartEVAL.Rd b/man/RpartEVAL.Rd index a1fc00e..5fcd1e0 100644 --- a/man/RpartEVAL.Rd +++ b/man/RpartEVAL.Rd @@ -28,15 +28,8 @@ This function evaluates the performance of the generated trees for error estimation by ten-fold cross validation assessment. } \examples{ -sc <- DISCBIO(valuesG1msReduced) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -sc <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, - dsn=1, rseed=17000 -) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -sc <- Clustexp(sc, cln=3) # K-means clustering -sc <- comptSNE(sc, max_iter=100) +data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +sc <- valuesG1msReduced_treated_K cdiff <- DEGanalysis2clust( sc, Clustering="K-means", K=3, fdr=.2, name="Name", First="CL1", Second="CL2", export=FALSE diff --git a/man/plotExptSNE.Rd b/man/plotExptSNE.Rd index 80dbb8a..1bcf8a3 100644 --- a/man/plotExptSNE.Rd +++ b/man/plotExptSNE.Rd @@ -28,9 +28,8 @@ The t-SNE map representation can also be used to analyze gene expression patterns } \examples{ -sc <- DISCBIO(valuesG1msReduced) -sc <- Clustexp(sc, cln=3, quiet=TRUE) # K-means clustering -sc <- comptSNE(sc, max_iter=100) +data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details +sc <- valuesG1msReduced_treated_K g <- 'ENSG00000001460' plotExptSNE(sc, g) } diff --git a/man/plotGap.Rd b/man/plotGap.Rd index e71f6c7..6fd97e6 100644 --- a/man/plotGap.Rd +++ b/man/plotGap.Rd @@ -21,7 +21,7 @@ A plot of the gap statistics Plotting Gap Statistics } \examples{ -sc <- DISCBIO(valuesG1msReduced) # changes signature of data -sc <- Clustexp(sc, cln=3) # data must be clustered before plotting +data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details +sc <- valuesG1msReduced_treated_K plotGap(sc) } diff --git a/man/plotKmeansLabelstSNE.Rd b/man/plotKmeansLabelstSNE.Rd index 82d3e3f..94153ee 100644 --- a/man/plotKmeansLabelstSNE.Rd +++ b/man/plotKmeansLabelstSNE.Rd @@ -19,8 +19,7 @@ Plot containing the ID of the cells in each cluster Visualizing the K-means clusters using tSNE maps } \examples{ -sc <- DISCBIO(valuesG1msReduced) # changes signature of data -sc <- Clustexp(sc, cln=3) # data must be clustered before plottin -sc <- comptSNE(sc, max_iter=100) +data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details +sc <- valuesG1msReduced_treated_K plotKmeansLabelstSNE(sc) # Plots the ID of the cells in each cluster } diff --git a/man/plotMBLabelstSNE.Rd b/man/plotMBLabelstSNE.Rd index 93585f5..014d2c0 100644 --- a/man/plotMBLabelstSNE.Rd +++ b/man/plotMBLabelstSNE.Rd @@ -19,14 +19,7 @@ A plot of the `object@MBtsne` values Visualizing the Model-based clusters using tSNE maps } \examples{ -sc <- DISCBIO(valuesG1msReduced) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -sc <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, - dsn=1, rseed=17000 -) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -sc <- Exprmclust(sc, K=2) -sc <- comptsneMB(sc, rseed=15555, quiet=TRUE, max_iter=100) +data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset +sc <- valuesG1msReduced_treated_MB plotMBLabelstSNE(sc) } diff --git a/man/plotOrderKMtsne.Rd b/man/plotOrderKMtsne.Rd index 0122164..81e1848 100644 --- a/man/plotOrderKMtsne.Rd +++ b/man/plotOrderKMtsne.Rd @@ -21,9 +21,8 @@ The tSNE representation can also be used to show the pseudo-time ordering. } \examples{ -sc <- DISCBIO(valuesG1msReduced) -sc <- Clustexp(sc, cln=3) # K-means clustering -sc <- comptSNE(sc, max_iter=100) +data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +sc <- valuesG1msReduced_treated_K sc <- KmeanOrder(sc, export = FALSE) plotOrderKMtsne(sc) } diff --git a/man/plotOrderMBtsne.Rd b/man/plotOrderMBtsne.Rd index 3d83b3f..bd93d9b 100644 --- a/man/plotOrderMBtsne.Rd +++ b/man/plotOrderMBtsne.Rd @@ -21,11 +21,7 @@ The tSNE representation can also be used to show the pseudo-time ordering. } \examples{ -sc<- DISCBIO(valuesG1msReduced) -sc<-NoiseFiltering(sc,percentile=0.9, CV=0.2, export=FALSE) -sc<-Normalizedata(sc) -sc<-FinalPreprocessing(sc,GeneFlitering="NoiseF", export=FALSE) -sc <- Exprmclust(sc, K=2, reduce=TRUE, quiet=TRUE) -sc<- comptsneMB(sc, rseed=15555, quiet=TRUE, max_iter=100) +data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset +sc <- valuesG1msReduced_treated_MB plotOrderMBtsne(sc) } diff --git a/man/plotSilhouette.Rd b/man/plotSilhouette.Rd index 0a88fd0..cea359c 100644 --- a/man/plotSilhouette.Rd +++ b/man/plotSilhouette.Rd @@ -31,8 +31,7 @@ The silhouette provides a representation of how well each point representation of a point by its cluster. } \examples{ -sc <- DISCBIO(valuesG1msReduced) # changes signature of data -sc <- Clustexp(sc, cln=3) # data must be clustered before plottin -sc <- comptSNE(sc, max_iter=100) +data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +sc <- valuesG1msReduced_treated_K plotSilhouette(sc, K=3) } diff --git a/man/plotSymbolstSNE.Rd b/man/plotSymbolstSNE.Rd index 9c520b7..f4a4036 100644 --- a/man/plotSymbolstSNE.Rd +++ b/man/plotSymbolstSNE.Rd @@ -26,8 +26,7 @@ Plot of tsne objet slot, grouped by gene. Visualizing the K-means clusters using tSNE maps } \examples{ -sc <- DISCBIO(valuesG1msReduced) # changes signature of data -sc <- Clustexp(sc, cln=3) # data must be clustered before plottin -sc <- comptSNE(sc, max_iter=100, quiet=TRUE) +data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +sc <- valuesG1msReduced_treated_K plotSymbolstSNE(sc,types=sub("(\\\\_\\\\d+)$","", names(sc@ndata))) } diff --git a/man/plotsilhouetteMB.Rd b/man/plotsilhouetteMB.Rd index c892fe8..477b329 100644 --- a/man/plotsilhouetteMB.Rd +++ b/man/plotsilhouetteMB.Rd @@ -27,13 +27,7 @@ The silhouette provides a representation of how well each point representation of a point by its cluster. } \examples{ -sc <- DISCBIO(valuesG1msReduced) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -sc <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, - downsample=FALSE, dsn=1, rseed=17000 -) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -sc <- Exprmclust(sc, K=2, reduce=TRUE, quiet=TRUE) +data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset +sc <- valuesG1msReduced_treated_MB plotsilhouetteMB(sc, K=2) } diff --git a/man/plottSNE.Rd b/man/plottSNE.Rd index 046c9f3..bf1c831 100644 --- a/man/plottSNE.Rd +++ b/man/plottSNE.Rd @@ -16,8 +16,7 @@ A plot of t-SNEs. Visualizing the K-means clusters using tSNE maps } \examples{ -sc <- DISCBIO(valuesG1msReduced) # changes signature of data -sc <- Clustexp(sc, cln=3) # data must be clustered before plotting -sc <- comptSNE(sc, max_iter=100, quiet=TRUE) +data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset +sc <- valuesG1msReduced_treated_K plottSNE(sc) } diff --git a/man/plottsneMB.Rd b/man/plottsneMB.Rd index 6e6a09b..5214204 100644 --- a/man/plottsneMB.Rd +++ b/man/plottsneMB.Rd @@ -21,14 +21,7 @@ A plot of t-SNEs. Visualizing the Model-based clusters using tSNE maps } \examples{ -sc <- DISCBIO(valuesG1msReduced) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -sc <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, - dsn=1, rseed=17000 -) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -sc <- Exprmclust(sc, K=2) -sc <- comptsneMB(sc, rseed=15555, quiet=TRUE, max_iter=100) +data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset +sc <- valuesG1msReduced_treated_MB plottsneMB(sc) } From bf53b6839d1f60e0e7a81df7e5eaa193bd34fa30 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Thu, 2 Jul 2020 10:24:17 +0200 Subject: [PATCH 30/99] Further reduced examples --- R/DIscBIO-generic-comptSNE.R | 2 +- man/comptSNE.Rd | 2 +- 2 files changed, 2 insertions(+), 2 deletions(-) diff --git a/R/DIscBIO-generic-comptSNE.R b/R/DIscBIO-generic-comptSNE.R index dcb9ea3..a80d982 100644 --- a/R/DIscBIO-generic-comptSNE.R +++ b/R/DIscBIO-generic-comptSNE.R @@ -14,7 +14,7 @@ #' @examples #' sc <- DISCBIO(valuesG1msReduced) # changes signature of data #' sc <- Clustexp(sc, cln=3) # data must be clustered before plottin -#' sc <- comptSNE(sc, max_iter=1000) +#' sc <- comptSNE(sc, max_iter=30) #' head(sc@tsne) setGeneric( name = "comptSNE", diff --git a/man/comptSNE.Rd b/man/comptSNE.Rd index 52308b3..c3e56b3 100644 --- a/man/comptSNE.Rd +++ b/man/comptSNE.Rd @@ -47,6 +47,6 @@ This function is used to compute the t-Distributed Stochastic \examples{ sc <- DISCBIO(valuesG1msReduced) # changes signature of data sc <- Clustexp(sc, cln=3) # data must be clustered before plottin -sc <- comptSNE(sc, max_iter=1000) +sc <- comptSNE(sc, max_iter=30) head(sc@tsne) } From fdb41638cb58fe718816b54269684238ec20ebae Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Thu, 2 Jul 2020 11:24:35 +0200 Subject: [PATCH 31/99] Added reset code to par() and options() changes --- R/DIscBIO-generic-Clustexp.R | 9 ++++++--- R/DIscBIO-generic-KMclustheatmap.R | 6 ++++-- R/DIscBIO-generic-MBclustheatmap.R | 6 ++++-- R/DIscBIO-generic-plotExptSNE.R | 6 ++++-- R/DIscBIO-generic-plotOrderKMtsne.R | 6 ++++-- R/DIscBIO-generic-plotOrderMBtsne.R | 6 ++++-- R/DIscBIO-generic-plotexptsneMB.R | 6 ++++-- R/Networking.R | 7 +++++-- R/PlotMBexpPCA.R | 6 ++++-- R/PlotMBorderPCA.R | 6 ++++-- vignettes/DIscBIO-vignette.Rmd | 4 ++-- 11 files changed, 45 insertions(+), 23 deletions(-) diff --git a/R/DIscBIO-generic-Clustexp.R b/R/DIscBIO-generic-Clustexp.R index cd1f8a2..f2bf8db 100644 --- a/R/DIscBIO-generic-Clustexp.R +++ b/R/DIscBIO-generic-Clustexp.R @@ -205,7 +205,8 @@ setMethod( kmopt <- NULL for (i in 1:runs) { - options(show.error.messages = FALSE) + opar <- options(show.error.messages = FALSE) + on.exit(par(opar)) repeat { kmm <- try(Kmeans( data, @@ -218,7 +219,8 @@ setMethod( "try-error")) break } - options(show.error.messages = TRUE) + opar <- options(show.error.messages = TRUE) + on.exit(par(opar)) swss <- sum(kmm$withinss) if (swss < minSS) { @@ -227,7 +229,8 @@ setMethod( swss } if (plot) { - par(ask = TRUE) + opar <- par(ask = TRUE) + on.exit(par(opar)) pairs(data, col = kmm$cluster, main = swss) diff --git a/R/DIscBIO-generic-KMclustheatmap.R b/R/DIscBIO-generic-KMclustheatmap.R index 7ee5196..f398445 100644 --- a/R/DIscBIO-generic-KMclustheatmap.R +++ b/R/DIscBIO-generic-KMclustheatmap.R @@ -109,7 +109,8 @@ setMethod( ) } - par(mar = c(3, 5, 2.5, 2)) + opar <- par(mar = c(3, 5, 2.5, 2)) + on.exit(par(opar)) image(as.matrix(di[ptn, ptn]), col = ColorRamp, axes = FALSE) abline(0, 1) box() @@ -136,7 +137,8 @@ setMethod( } axis(1, at = tmp, labels = cclmo) axis(2, at = tmp, labels = cclmo) - par(mar = c(3, 2.5, 2.5, 2)) + opar <- par(mar = c(3, 2.5, 2.5, 2)) + on.exit(par(opar)) image( 1, ColorLevels, diff --git a/R/DIscBIO-generic-MBclustheatmap.R b/R/DIscBIO-generic-MBclustheatmap.R index a41c005..47fb31c 100644 --- a/R/DIscBIO-generic-MBclustheatmap.R +++ b/R/DIscBIO-generic-MBclustheatmap.R @@ -191,7 +191,8 @@ setMethod( if (mi == ma) { ColorLevels <- seq(0.99 * mi, 1.01 * ma, length = length(ColorRamp)) } - par(mar = c(3, 5, 2.5, 2)) + opar <- par(mar = c(3, 5, 2.5, 2)) + on.exit(par(opar)) image(as.matrix(di[ptn, ptn]), col = ColorRamp, axes = FALSE) abline(0, 1) box() @@ -218,7 +219,8 @@ setMethod( } axis(1, at = tmp, labels = cclmo) axis(2, at = tmp, labels = cclmo) - par(mar = c(3, 2.5, 2.5, 2)) + opar <- par(mar = c(3, 2.5, 2.5, 2)) + on.exit(par(opar)) image( 1, ColorLevels, diff --git a/R/DIscBIO-generic-plotExptSNE.R b/R/DIscBIO-generic-plotExptSNE.R index 1f8e6f5..89aba63 100644 --- a/R/DIscBIO-generic-plotExptSNE.R +++ b/R/DIscBIO-generic-plotExptSNE.R @@ -50,7 +50,8 @@ setMethod( widths = c(5, 1, 5, 1), heights = c(5, 1, 1, 1) ) - par(mar = c(3, 5, 2.5, 2)) + opar <- par(mar = c(3, 5, 2.5, 2)) + on.exit(par(opar)) plot( object@tsne, xlab = "Dim 1", @@ -70,7 +71,8 @@ setMethod( cex = 1.5 ) } - par(mar = c(3, 2.5, 2.5, 2)) + opar <- par(mar = c(3, 2.5, 2.5, 2)) + on.exit(par(opar)) image( 1, ColorLevels, diff --git a/R/DIscBIO-generic-plotOrderKMtsne.R b/R/DIscBIO-generic-plotOrderKMtsne.R index ce5e946..1716a4e 100644 --- a/R/DIscBIO-generic-plotOrderKMtsne.R +++ b/R/DIscBIO-generic-plotOrderKMtsne.R @@ -42,7 +42,8 @@ setMethod( widths = c(5, 1, 5, 1), heights = c(5, 1, 1, 1) ) - par(mar = c(3, 5, 2.5, 2)) + opar <- par(mar = c(3, 5, 2.5, 2)) + on.exit(par(opar)) plot( object@tsne, xlab = "Dim 1", @@ -62,7 +63,8 @@ setMethod( cex = 1.5 ) } - par(mar = c(3, 2.5, 2.5, 2)) + opar <- par(mar = c(3, 2.5, 2.5, 2)) + on.exit(par(opar)) image( 1, ColorLevels, diff --git a/R/DIscBIO-generic-plotOrderMBtsne.R b/R/DIscBIO-generic-plotOrderMBtsne.R index 26d4388..4a9aa0e 100644 --- a/R/DIscBIO-generic-plotOrderMBtsne.R +++ b/R/DIscBIO-generic-plotOrderMBtsne.R @@ -41,7 +41,8 @@ setMethod( widths = c(5, 1, 5, 1), heights = c(5, 1, 1, 1) ) - par(mar = c(3, 5, 2.5, 2)) + opar <- par(mar = c(3, 5, 2.5, 2)) + on.exit(par(opar)) plot( object@MBtsne, xlab = "Dim 1", @@ -61,7 +62,8 @@ setMethod( cex = 1.5 ) } - par(mar = c(3, 2.5, 2.5, 2)) + opar <- par(mar = c(3, 2.5, 2.5, 2)) + on.exit(par(opar)) image( 1, ColorLevels, diff --git a/R/DIscBIO-generic-plotexptsneMB.R b/R/DIscBIO-generic-plotexptsneMB.R index e37436f..e729561 100644 --- a/R/DIscBIO-generic-plotexptsneMB.R +++ b/R/DIscBIO-generic-plotexptsneMB.R @@ -53,7 +53,8 @@ setMethod( widths = c(5, 1, 5, 1), heights = c(5, 1, 1, 1) ) - par(mar = c(3, 5, 2.5, 2)) + opar <- par(mar = c(3, 5, 2.5, 2)) + on.exit(par(opar)) plot( object@MBtsne, xlab = "Dim 1", @@ -73,7 +74,8 @@ setMethod( cex = 1.5 ) } - par(mar = c(3, 2.5, 2.5, 2)) + opar <- par(mar = c(3, 2.5, 2.5, 2)) + on.exit(par(opar)) image( 1, ColorLevels, diff --git a/R/Networking.R b/R/Networking.R index 1afbb10..bc9c686 100644 --- a/R/Networking.R +++ b/R/Networking.R @@ -67,8 +67,11 @@ Networking <- function( Network <- readPNG(FileName, native=TRUE) set_plot_dimensions <- function(width_choice, height_choice) { - options(repr.plot.width = width_choice, - repr.plot.height = height_choice) + opar <- options( + repr.plot.width = width_choice, + repr.plot.height = height_choice + ) + on.exit(par(opar)) } set_plot_dimensions(plot_width, plot_height) diff --git a/R/PlotMBexpPCA.R b/R/PlotMBexpPCA.R index 7909499..30bf9d0 100644 --- a/R/PlotMBexpPCA.R +++ b/R/PlotMBexpPCA.R @@ -45,7 +45,8 @@ PlotMBexpPCA <- function(object, g, n = NULL) { widths = c(5, 1, 5, 1), heights = c(5, 1, 1, 1) ) - par(mar = c(5, 5, 2.5, 2)) + opar <- par(mar = c(5, 5, 2.5, 2)) + on.exit(par(opar)) plot( data$pcareduceres[, 1], data$pcareduceres[, 2], @@ -66,7 +67,8 @@ PlotMBexpPCA <- function(object, g, n = NULL) { cex = 2 ) } - par(mar = c(3, 2.5, 2.5, 2)) + opar <- par(mar = c(3, 2.5, 2.5, 2)) + on.exit(par(opar)) image( 1, ColorLevels, diff --git a/R/PlotMBorderPCA.R b/R/PlotMBorderPCA.R index ef91613..58c6523 100644 --- a/R/PlotMBorderPCA.R +++ b/R/PlotMBorderPCA.R @@ -32,7 +32,8 @@ PlotMBorderPCA <- function(object) { widths = c(5, 1, 5, 1), heights = c(5, 1, 1, 1) ) - par(mar = c(5, 5, 2.5, 2)) + opar <- par(mar = c(5, 5, 2.5, 2)) + on.exit(par(opar)) plot( MBordertable[, 1], MBordertable[, 2], @@ -52,7 +53,8 @@ PlotMBorderPCA <- function(object) { cex = 2 ) } - par(mar = c(3, 2.5, 2.5, 2)) + opar <- par(mar = c(3, 2.5, 2.5, 2)) + on.exit(par(opar)) image( 1, ColorLevels, diff --git a/vignettes/DIscBIO-vignette.Rmd b/vignettes/DIscBIO-vignette.Rmd index 06bdba8..5ec169f 100644 --- a/vignettes/DIscBIO-vignette.Rmd +++ b/vignettes/DIscBIO-vignette.Rmd @@ -164,7 +164,7 @@ PSCAN enables the robustness assessment of the detected clusters in terms of sta ```{r} # Silhouette of k-means clusters -par(mar=c(6,2,4,2)) +# you might have to run "par(mar=c(6,2,4,2))" first so plot looks better plotSilhouette(sc,K=2) # K is the number of clusters ``` @@ -453,7 +453,7 @@ plotMBLabelstSNE(sc) ```{r, eval=FALSE} # Silhouette of Model-based clusters -par(mar=c(6,2,4,2)) +# you might have to run "par(mar=c(6,2,4,2))" first so plot looks better plotsilhouetteMB(sc,K=3) ``` From 0ebaf8e501346033b50b1a8df8ba56d6978b570b Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Thu, 2 Jul 2020 12:18:38 +0200 Subject: [PATCH 32/99] Added filename arguments to exporting functions --- R/DIscBIO-generic-DEGanalysis.R | 58 +++++++++++++++------------ R/DIscBIO-generic-DEGanalysis2clust.R | 16 ++++---- R/DIscBIO-generic-KMClustDiffGenes.R | 32 +++++++++++---- R/DIscBIO-generic-KmeanOrder.R | 18 ++++----- R/DIscBIO-generic-MBClustDiffGenes.R | 34 +++++++++++----- R/DIscBIO-generic-NoiseFiltering.R | 8 ++-- R/MB_Order.R | 12 +++--- R/NetAnalysis.R | 8 ++-- R/PPI.R | 2 +- man/DEGanalysis.Rd | 16 ++++++-- man/DEGanalysis2clust.Rd | 16 ++++++-- man/KMClustDiffGenes.Rd | 20 ++++++++- man/KmeanOrder.Rd | 16 +++++++- man/MBClustDiffGenes.Rd | 20 ++++++++- man/MB_Order.Rd | 9 ++++- man/NetAnalysis.Rd | 2 +- man/NoiseFiltering.Rd | 8 +++- 17 files changed, 203 insertions(+), 92 deletions(-) diff --git a/R/DIscBIO-generic-DEGanalysis.R b/R/DIscBIO-generic-DEGanalysis.R index 57384a1..b663909 100644 --- a/R/DIscBIO-generic-DEGanalysis.R +++ b/R/DIscBIO-generic-DEGanalysis.R @@ -13,6 +13,8 @@ #' excel file. Default is TRUE. #' @param quiet if `TRUE`, suppresses intermediate text output #' @param plot if `TRUE`, plots are generated +#' @param filename_deg Name of the exported DEG table +#' @param filename_sigdeg Name of the exported sigDEG table #' @importFrom samr samr samr.compute.delta.table samr.plot #' samr.compute.siggenes.table #' @importFrom graphics title @@ -28,16 +30,20 @@ #' ) setGeneric( name = "DEGanalysis", - def = function(object, - Clustering = "K-means", - K, - fdr = 0.05, - name = "Name", - export = TRUE, - quiet = FALSE, - plot = TRUE, - ...) - standardGeneric("DEGanalysis") + def = function( + object, + K, + Clustering = "K-means", + fdr = 0.05, + name = "Name", + export = TRUE, + quiet = FALSE, + plot = TRUE, + filename_deg = "DEGsTable", + filename_sigdeg = "sigDEG", + ...) { + standardGeneric("DEGanalysis") + } ) #' @export @@ -45,16 +51,19 @@ setGeneric( setMethod( f = "DEGanalysis", signature = "DISCBIO", - definition = function(object, - Clustering = "K-means", - K, - fdr = 0.05, - name = "Name", - export = TRUE, - quiet = FALSE, - plot = TRUE, - ...) - { + definition = function( + object, + K, + Clustering = "K-means", + fdr = 0.05, + name = "Name", + export = TRUE, + quiet = FALSE, + plot = TRUE, + filename_deg, + filename_sigdeg, + ... + ) { # Validation if (!(Clustering %in% c("K-means", "MB"))) { stop("Clustering has to be either K-means or MB") @@ -444,13 +453,10 @@ setMethod( "Comparisons", "Target cluster", "Gene number", "File name", "Gene number", "File name" ) - if (export) - write.csv(DEGsTable, file = "DEGsTable.csv") - if (!quiet) - print(DEGsTable) + if (export) write.csv(DEGsTable, file = paste0(filename_deg, ".csv")) + if (!quiet) print(DEGsTable) sigDEG <- cbind(DEGsE, DEGsS) - if (export) - write.csv(sigDEG, file = "sigDEG.csv") + if (export) write.csv(sigDEG, file = paste0(filename_sigdeg, ".csv")) return(list(sigDEG, DEGsTable)) } ) \ No newline at end of file diff --git a/R/DIscBIO-generic-DEGanalysis2clust.R b/R/DIscBIO-generic-DEGanalysis2clust.R index 536b7d7..beccd9c 100644 --- a/R/DIscBIO-generic-DEGanalysis2clust.R +++ b/R/DIscBIO-generic-DEGanalysis2clust.R @@ -17,6 +17,8 @@ #' excel file. Default is TRUE. #' @param quiet if `TRUE`, suppresses intermediate text output #' @param plot if `TRUE`, plots are generated +#' @param filename_deg Name of the exported DEG table +#' @param filename_sigdeg Name of the exported sigDEG table #' @param ... additional parameters to be passed to samr() #' @importFrom samr samr samr.compute.delta.table samr.plot #' samr.compute.siggenes.table @@ -33,9 +35,10 @@ setGeneric( "DEGanalysis2clust", function( - object, Clustering = "K-means", K, fdr = 0.05, name = "Name", + object, K, Clustering = "K-means", fdr = 0.05, name = "Name", First = "CL1", Second = "CL2", export = TRUE, quiet = FALSE, - plot = TRUE, ... + plot = TRUE, filename_deg = "DEGsTable", filename_sigdeg = "sigDEG", + ... ) standardGeneric("DEGanalysis2clust") ) @@ -46,9 +49,8 @@ setMethod( "DEGanalysis2clust", signature = "DISCBIO", definition = function( - object, Clustering = "K-means", K, fdr = 0.05, name = "Name", - First = "CL1", Second = "CL2", export = TRUE, quiet = FALSE, - plot = TRUE, ...) + object, K, Clustering, fdr, name, First, Second, export, quiet, plot, + filename_deg, filename_sigdeg, ...) { if (!(Clustering %in% c("K-means", "MB"))) { stop("Clustering has to be either K-means or MB") @@ -331,8 +333,8 @@ setMethod( if (!quiet) print(DEGsTable) sigDEG <- cbind(DEGsE, DEGsS) if (export) { - write.csv(DEGsTable, file = "DEGsTable.csv") - write.csv(sigDEG, file = "sigDEG.csv") + write.csv(DEGsTable, file = paste0(filename_deg, ".csv")) + write.csv(sigDEG, file = paste0(filename_sigdeg, ".csv")) } return( list( diff --git a/R/DIscBIO-generic-KMClustDiffGenes.R b/R/DIscBIO-generic-KMClustDiffGenes.R index d516fb3..03ee8c5 100644 --- a/R/DIscBIO-generic-KMClustDiffGenes.R +++ b/R/DIscBIO-generic-KMClustDiffGenes.R @@ -7,6 +7,10 @@ #' @param export A logical vector that allows writing the final gene list in #' excel file. Default is TRUE. #' @param quiet if `TRUE`, suppresses intermediate text output +#' @param filename_up Name of the exported "up" file (if `export=TRUE`) +#' @param filename_down Name of the exported "down" file (if `export=TRUE`) +#' @param filename_binom Name of the exported binomial file +#' @param filename_sigdeg Name of the exported sigDEG file #' @importFrom dplyr summarize #' @importFrom stats pbinom median #' @rdname KMClustDiffGenes @@ -18,15 +22,25 @@ #' KMClustDiffGenes(sc, K=3, fdr=.3, export=FALSE) setGeneric( "KMClustDiffGenes", - function(object, K, pValue = 0.05, fdr = .01, export = TRUE, quiet = FALSE) + function( + object, K, pValue = 0.05, fdr = .01, export = TRUE, quiet = FALSE, + filename_up = "Up-DEG-cluster", + filename_down = "Down-DEG-cluster", + filename_binom = "binomial-DEGsTable", + filename_sigdeg = "binomial-sigDEG" + ) { standardGeneric("KMClustDiffGenes") + } ) #' @export #' @rdname KMClustDiffGenes setMethod( "KMClustDiffGenes", signature = "DISCBIO", - definition = function(object, K, pValue, fdr, export, quiet) { + definition = function( + object, K, pValue, fdr, export, quiet, filename_up, filename_down, + filename_binom, filename_sigdeg + ) { # Validation if (!is.numeric(fdr)) { stop("fdr has to be a number between 0 and 1") @@ -166,7 +180,7 @@ setMethod( ) if (export) { write.csv( - Up, file = paste0("Up-DEG-cluster", n, ".csv") + Up, file = paste0(filename_up, n, ".csv") ) } @@ -190,21 +204,23 @@ setMethod( if (export) { write.csv( Down, - file = paste0("Down-DEG-cluster", n, ".csv") + file = paste0(filename_down, n, ".csv") ) } sigDEG <- cbind(DEGsE, DEGsS) if (export) { - write.csv(sigDEG, file = "binomial-sigDEG.csv") + write.csv( + sigDEG, file = paste0(filename_sigdeg, ".csv") + ) } DEGsTable[n, 1] <- paste0("Cluster ", n) DEGsTable[n, 2] <- "Remaining Clusters" DEGsTable[n, 3] <- length(Up[, 1]) - DEGsTable[n, 4] <- paste0("Up-DEG-cluster", n, ".csv") + DEGsTable[n, 4] <- paste0(filename_up, n, ".csv") DEGsTable[n, 5] <- length(Down[, 1]) - DEGsTable[n, 6] <- paste0("Down-DEG-cluster", n, ".csv") + DEGsTable[n, 6] <- paste0(filename_down, n, ".csv") } } } @@ -214,7 +230,7 @@ setMethod( "Gene number", "File name" ) if (export) { - write.csv(DEGsTable, file = "binomial-DEGsTable.csv") + write.csv(DEGsTable, file = paste0(filename_binom, ".csv")) } return(list(sigDEG, DEGsTable)) } else{ diff --git a/R/DIscBIO-generic-KmeanOrder.R b/R/DIscBIO-generic-KmeanOrder.R index 96ee938..a4061a0 100644 --- a/R/DIscBIO-generic-KmeanOrder.R +++ b/R/DIscBIO-generic-KmeanOrder.R @@ -5,6 +5,7 @@ #' @param object \code{DISCBIO} class object. #' @param quiet if `TRUE`, suppresses intermediary output #' @param export if `TRUE`, exports order table to csv +#' @param filename Name of the exported file (if `export=TRUE`) #' @importFrom TSCAN TSCANorder #' @return The DISCBIO-class object input with the kordering slot filled. #' @examples @@ -12,7 +13,8 @@ #' sc <- valuesG1msReduced_treated_K #' Order <- KmeanOrder(sc, export = FALSE) #' Order@kordering -setGeneric("KmeanOrder", function(object, quiet = FALSE, export = TRUE) +setGeneric("KmeanOrder", function(object, quiet = FALSE, export = TRUE, + filename = "Cellular_pseudo-time_ordering_based_on_k-meansc-lusters") standardGeneric("KmeanOrder") ) @@ -21,7 +23,7 @@ setGeneric("KmeanOrder", function(object, quiet = FALSE, export = TRUE) setMethod( "KmeanOrder", signature = "DISCBIO", - definition = function(object, quiet = FALSE, export = TRUE) { + definition = function(object, quiet, export, filename) { # Validation if (length(object@kmeans$kpart) == 0) { stop("run Clustexp before KmeanOrder") @@ -35,15 +37,9 @@ setMethod( orderID <- lpsorder order <- c(1:length(lpsorder)) orderTable <- data.frame(order, orderID) - if (export) { - nm <- "Cellular_pseudo-time_ordering_based_on_k-meansc-lusters.csv" - write.csv(orderTable, file = nm) - } - if (!quiet) { - print(orderTable) - } - FinalOrder <- - orderTable[match(sampleNames, orderTable$orderID), ] + if (export) write.csv(orderTable, file = paste0(filename, ".csv")) + if (!quiet) print(orderTable) + FinalOrder <- orderTable[match(sampleNames, orderTable$orderID), ] out_order <- FinalOrder[, 1] names(out_order) <- names(Clusters) object@kordering <- out_order diff --git a/R/DIscBIO-generic-MBClustDiffGenes.R b/R/DIscBIO-generic-MBClustDiffGenes.R index 02f7ac1..1ce620f 100644 --- a/R/DIscBIO-generic-MBClustDiffGenes.R +++ b/R/DIscBIO-generic-MBClustDiffGenes.R @@ -7,6 +7,10 @@ #' @param export A logical vector that allows writing the final gene list in #' excel file. Default is TRUE. #' @param quiet if `TRUE`, suppresses intermediate text output +#' @param filename_up Name of the exported "up" file (if `export=TRUE`) +#' @param filename_down Name of the exported "down" file (if `export=TRUE`) +#' @param filename_binom Name of the exported binomial file +#' @param filename_sigdeg Name of the exported sigDEG file #' @importFrom stats pbinom median #' @importFrom AnnotationDbi select #' @import org.Hs.eg.db @@ -22,15 +26,25 @@ #' } setGeneric( "MBClustDiffGenes", - function(object, K, pValue = 0.05, fdr = .01, export = TRUE, quiet = FALSE) - standardGeneric("MBClustDiffGenes") + function( + object, K, pValue = 0.05, fdr = .01, export = TRUE, quiet = FALSE, + filename_up = "Up-DEG-cluster", + filename_down = "Down-DEG-cluster", + filename_binom = "binomial-DEGsTable", + filename_sigdeg = "binomial-sigDEG" + ) { + standardGeneric("MBClustDiffGenes") + } ) #' @export #' @rdname MBClustDiffGenes setMethod( "MBClustDiffGenes", signature = "DISCBIO", - definition = function(object, K, pValue, fdr, export, quiet) { + definition = function( + object, K, pValue, fdr, export, quiet, filename_up, filename_down, + filename_binom, filename_sigdeg + ) { if (!is.numeric(fdr)) { stop("fdr has to be a number between 0 and 1") } else if (fdr < 0 | fdr > 1) { @@ -162,7 +176,7 @@ setMethod( ) if (export) { write.csv( - Up, file = paste0("Up-DEG-cluster", n, ".csv") + Up, file = paste0(filename_up, n, ".csv") ) } @@ -186,23 +200,25 @@ setMethod( if (export) { write.csv( Down, - file = paste0("Down-DEG-cluster", n, ".csv") + file = paste0(filename_down, n, ".csv") ) } sigDEG <- cbind(DEGsE, DEGsS) if (export) { - write.csv(sigDEG, file = "binomial-sigDEG.csv") + write.csv( + sigDEG, file = paste0(filename_sigdeg, ".csv") + ) } DEGsTable[n, 1] <- paste0("Cluster ", n) DEGsTable[n, 2] <- "Remaining Clusters" DEGsTable[n, 3] <- length(Up[, 1]) DEGsTable[n, 4] <- - paste0("Up-DEG-cluster", n, ".csv") + paste0(filename_up, n, ".csv") DEGsTable[n, 5] <- length(Down[, 1]) DEGsTable[n, 6] <- - paste0("Down-DEG-cluster", n, ".csv") + paste0(filename_down, n, ".csv") } } } @@ -212,7 +228,7 @@ setMethod( "Gene number", "File name" ) if (export) { - write.csv(DEGsTable, file = "binomial-DEGsTable.csv") + write.csv(DEGsTable, file = paste0(filename_binom, ".csv")) } return(list(sigDEG, DEGsTable)) } else{ diff --git a/R/DIscBIO-generic-NoiseFiltering.R b/R/DIscBIO-generic-NoiseFiltering.R index 97c602d..e25046c 100644 --- a/R/DIscBIO-generic-NoiseFiltering.R +++ b/R/DIscBIO-generic-NoiseFiltering.R @@ -19,6 +19,7 @@ #' @param export A logical vector that allows writing the final gene list in #' excel file. Default is TRUE. #' @param quiet if `TRUE`, suppresses printed output +#' @param filename Name of the exported file (if `export=TRUE`) #' @importFrom matrixStats rowVars #' @importFrom stats quantile var fitted.values pchisq p.adjust median aggregate #' @importFrom graphics plot axis abline points lines @@ -34,7 +35,8 @@ setGeneric( def = function( object, percentile = 0.8, CV = 0.3, geneCol = "yellow", FgeneCol = "black", erccCol = "blue", Val = TRUE, plot = TRUE, - export = TRUE, quiet = FALSE + export = TRUE, quiet = FALSE, + filename = "Noise_filtering_genes_test" ) standardGeneric("NoiseFiltering") ) @@ -46,7 +48,7 @@ setMethod( signature = "DISCBIO", definition = function( object, percentile, CV, geneCol, FgeneCol, erccCol, Val, plot, - export, quiet + export, quiet, filename ) { if (!is.numeric(percentile)) @@ -161,7 +163,7 @@ setMethod( } if (export) { - write.csv(genes_test, file = "Noise_filtering_genes_test.csv") + write.csv(genes_test, file = paste0(filename, ".csv")) cat( "The filtered gene list was saved as:", "Noise_filtering_genes_test\n" diff --git a/R/MB_Order.R b/R/MB_Order.R index 87b56aa..96e51b1 100644 --- a/R/MB_Order.R +++ b/R/MB_Order.R @@ -6,6 +6,7 @@ #' @param object \code{DISCBIO} class object. #' @param quiet if `TRUE`, intermediary output is suppressed #' @param export if `TRUE`, exports the results as a CSV file +#' @param filename Name of the exported file (if `export=TRUE`) #' @importFrom TSCAN TSCANorder #' @return The DISCBIO-class object input with the MBordering slot filled. #' @examples @@ -15,7 +16,10 @@ #' sc <- MB_Order(sc, export = FALSE) #' sc@MBordering #' } -MB_Order <- function(object, quiet = FALSE, export = TRUE) { +MB_Order <- function( + object, quiet = FALSE, export = TRUE, + filename = "Cellular_pseudo-time_ordering_based_on_Model-based_clusters" +) { data = object@MBclusters lpsorderMB <- TSCANorder(data) Names <- names(object@MBclusters$clusterid) @@ -24,14 +28,12 @@ MB_Order <- function(object, quiet = FALSE, export = TRUE) { order <- c(1:length(lpsorderMB)) orderTableMB <- data.frame(order, orderID) if (export) { - nm <- "Cellular_pseudo-time_ordering_based_on_Model-based_clusters.csv" - write.csv(orderTableMB, file = nm) + write.csv(orderTableMB, file = paste0(filename, ".csv")) } if (!quiet) { print(orderTableMB) } - FinalOrder <- - orderTableMB[match(sampleNames, orderTableMB$orderID), ] + FinalOrder <- orderTableMB[match(sampleNames, orderTableMB$orderID), ] MBordering <- FinalOrder[, 1] names(MBordering) <- names(Names) object@MBordering <- MBordering diff --git a/R/NetAnalysis.R b/R/NetAnalysis.R index 639a857..f024b82 100644 --- a/R/NetAnalysis.R +++ b/R/NetAnalysis.R @@ -19,7 +19,9 @@ #' ppi <- PPI(dff$FinalDEGsU[, "Gene Name"]) #' NetAnalysis(ppi, export=FALSE) #' } -NetAnalysis <- function(data, export = TRUE, FileName = "1") { +NetAnalysis <- function( + data, export = TRUE, FileName = "NetworkAnalysisTable-1" +) { if (length(data[, 1]) < 1) stop("No Protein-Protein Interactions") df <- data[, -c(1, 2)] @@ -35,9 +37,7 @@ NetAnalysis <- function(data, export = TRUE, FileName = "1") { AnalysisTable <- cbind(names, degree.table, betweenness.table) if (export) { - write.csv( - AnalysisTable, - file = paste0("NetworkAnalysisTable-", FileName, ".csv") + write.csv(AnalysisTable, file = paste0(FileName, ".csv") ) } diff --git a/R/PPI.R b/R/PPI.R index c0d8d97..1e5eef1 100644 --- a/R/PPI.R +++ b/R/PPI.R @@ -49,7 +49,7 @@ PPI <- function(data, FileName = NULL, species = "9606") { ) # Process API request content repo_content <- content(repos) - results <- read_tsv(repo_content) + results <- read_tsv(repo_content) if (!is.null(FileName)) { write.csv(results, file = paste0("PPI-", FileName, ".csv")) } diff --git a/man/DEGanalysis.Rd b/man/DEGanalysis.Rd index ba717ad..0c1798f 100644 --- a/man/DEGanalysis.Rd +++ b/man/DEGanalysis.Rd @@ -8,36 +8,40 @@ \usage{ DEGanalysis( object, - Clustering = "K-means", K, + Clustering = "K-means", fdr = 0.05, name = "Name", export = TRUE, quiet = FALSE, plot = TRUE, + filename_deg = "DEGsTable", + filename_sigdeg = "sigDEG", ... ) \S4method{DEGanalysis}{DISCBIO}( object, - Clustering = "K-means", K, + Clustering = "K-means", fdr = 0.05, name = "Name", export = TRUE, quiet = FALSE, plot = TRUE, + filename_deg = "DEGsTable", + filename_sigdeg = "sigDEG", ... ) } \arguments{ \item{object}{\code{DISCBIO} class object.} +\item{K}{A numeric value of the number of clusters.} + \item{Clustering}{Clustering has to be one of the following: ["K-means","MB"]. Default is "K-means"} -\item{K}{A numeric value of the number of clusters.} - \item{fdr}{A numeric value of the false discovery rate. Default is 0.05.} \item{name}{A string vector showing the name to be used to save the resulted @@ -50,6 +54,10 @@ excel file. Default is TRUE.} \item{plot}{if `TRUE`, plots are generated} +\item{filename_deg}{Name of the exported DEG table} + +\item{filename_sigdeg}{Name of the exported sigDEG table} + \item{...}{additional parameters to be passed to samr()} } \value{ diff --git a/man/DEGanalysis2clust.Rd b/man/DEGanalysis2clust.Rd index 9a2d461..2bc3921 100644 --- a/man/DEGanalysis2clust.Rd +++ b/man/DEGanalysis2clust.Rd @@ -8,8 +8,8 @@ \usage{ DEGanalysis2clust( object, - Clustering = "K-means", K, + Clustering = "K-means", fdr = 0.05, name = "Name", First = "CL1", @@ -17,13 +17,15 @@ DEGanalysis2clust( export = TRUE, quiet = FALSE, plot = TRUE, + filename_deg = "DEGsTable", + filename_sigdeg = "sigDEG", ... ) \S4method{DEGanalysis2clust}{DISCBIO}( object, - Clustering = "K-means", K, + Clustering = "K-means", fdr = 0.05, name = "Name", First = "CL1", @@ -31,17 +33,19 @@ DEGanalysis2clust( export = TRUE, quiet = FALSE, plot = TRUE, + filename_deg = "DEGsTable", + filename_sigdeg = "sigDEG", ... ) } \arguments{ \item{object}{\code{DISCBIO} class object.} +\item{K}{A numeric value of the number of clusters.} + \item{Clustering}{Clustering has to be one of the following: ["K-means","MB"]. Default is "K-means"} -\item{K}{A numeric value of the number of clusters.} - \item{fdr}{A numeric value of the false discovery rate. Default is 0.05.} \item{name}{A string vector showing the name to be used to save the resulted @@ -60,6 +64,10 @@ excel file. Default is TRUE.} \item{plot}{if `TRUE`, plots are generated} +\item{filename_deg}{Name of the exported DEG table} + +\item{filename_sigdeg}{Name of the exported sigDEG table} + \item{...}{additional parameters to be passed to samr()} } \value{ diff --git a/man/KMClustDiffGenes.Rd b/man/KMClustDiffGenes.Rd index e33d9f6..e6fd028 100644 --- a/man/KMClustDiffGenes.Rd +++ b/man/KMClustDiffGenes.Rd @@ -11,7 +11,11 @@ KMClustDiffGenes( pValue = 0.05, fdr = 0.01, export = TRUE, - quiet = FALSE + quiet = FALSE, + filename_up = "Up-DEG-cluster", + filename_down = "Down-DEG-cluster", + filename_binom = "binomial-DEGsTable", + filename_sigdeg = "binomial-sigDEG" ) \S4method{KMClustDiffGenes}{DISCBIO}( @@ -20,7 +24,11 @@ KMClustDiffGenes( pValue = 0.05, fdr = 0.01, export = TRUE, - quiet = FALSE + quiet = FALSE, + filename_up = "Up-DEG-cluster", + filename_down = "Down-DEG-cluster", + filename_binom = "binomial-DEGsTable", + filename_sigdeg = "binomial-sigDEG" ) } \arguments{ @@ -36,6 +44,14 @@ KMClustDiffGenes( excel file. Default is TRUE.} \item{quiet}{if `TRUE`, suppresses intermediate text output} + +\item{filename_up}{Name of the exported "up" file (if `export=TRUE`)} + +\item{filename_down}{Name of the exported "down" file (if `export=TRUE`)} + +\item{filename_binom}{Name of the exported binomial file} + +\item{filename_sigdeg}{Name of the exported sigDEG file} } \value{ A list containing two tables. diff --git a/man/KmeanOrder.Rd b/man/KmeanOrder.Rd index 2674efe..056891c 100644 --- a/man/KmeanOrder.Rd +++ b/man/KmeanOrder.Rd @@ -5,9 +5,19 @@ \alias{KmeanOrder,DISCBIO-method} \title{Pseudo-time ordering based on k-means clusters} \usage{ -KmeanOrder(object, quiet = FALSE, export = TRUE) +KmeanOrder( + object, + quiet = FALSE, + export = TRUE, + filename = "Cellular_pseudo-time_ordering_based_on_k-meansc-lusters" +) -\S4method{KmeanOrder}{DISCBIO}(object, quiet = FALSE, export = TRUE) +\S4method{KmeanOrder}{DISCBIO}( + object, + quiet = FALSE, + export = TRUE, + filename = "Cellular_pseudo-time_ordering_based_on_k-meansc-lusters" +) } \arguments{ \item{object}{\code{DISCBIO} class object.} @@ -15,6 +25,8 @@ KmeanOrder(object, quiet = FALSE, export = TRUE) \item{quiet}{if `TRUE`, suppresses intermediary output} \item{export}{if `TRUE`, exports order table to csv} + +\item{filename}{Name of the exported file (if `export=TRUE`)} } \value{ The DISCBIO-class object input with the kordering slot filled. diff --git a/man/MBClustDiffGenes.Rd b/man/MBClustDiffGenes.Rd index c0a002d..491911a 100644 --- a/man/MBClustDiffGenes.Rd +++ b/man/MBClustDiffGenes.Rd @@ -11,7 +11,11 @@ MBClustDiffGenes( pValue = 0.05, fdr = 0.01, export = TRUE, - quiet = FALSE + quiet = FALSE, + filename_up = "Up-DEG-cluster", + filename_down = "Down-DEG-cluster", + filename_binom = "binomial-DEGsTable", + filename_sigdeg = "binomial-sigDEG" ) \S4method{MBClustDiffGenes}{DISCBIO}( @@ -20,7 +24,11 @@ MBClustDiffGenes( pValue = 0.05, fdr = 0.01, export = TRUE, - quiet = FALSE + quiet = FALSE, + filename_up = "Up-DEG-cluster", + filename_down = "Down-DEG-cluster", + filename_binom = "binomial-DEGsTable", + filename_sigdeg = "binomial-sigDEG" ) } \arguments{ @@ -36,6 +44,14 @@ MBClustDiffGenes( excel file. Default is TRUE.} \item{quiet}{if `TRUE`, suppresses intermediate text output} + +\item{filename_up}{Name of the exported "up" file (if `export=TRUE`)} + +\item{filename_down}{Name of the exported "down" file (if `export=TRUE`)} + +\item{filename_binom}{Name of the exported binomial file} + +\item{filename_sigdeg}{Name of the exported sigDEG file} } \value{ A list containing two tables. diff --git a/man/MB_Order.Rd b/man/MB_Order.Rd index 35adaca..b6f3e1f 100644 --- a/man/MB_Order.Rd +++ b/man/MB_Order.Rd @@ -4,7 +4,12 @@ \alias{MB_Order} \title{Pseudo-time ordering based on Model-based clusters} \usage{ -MB_Order(object, quiet = FALSE, export = TRUE) +MB_Order( + object, + quiet = FALSE, + export = TRUE, + filename = "Cellular_pseudo-time_ordering_based_on_Model-based_clusters" +) } \arguments{ \item{object}{\code{DISCBIO} class object.} @@ -12,6 +17,8 @@ MB_Order(object, quiet = FALSE, export = TRUE) \item{quiet}{if `TRUE`, intermediary output is suppressed} \item{export}{if `TRUE`, exports the results as a CSV file} + +\item{filename}{Name of the exported file (if `export=TRUE`)} } \value{ The DISCBIO-class object input with the MBordering slot filled. diff --git a/man/NetAnalysis.Rd b/man/NetAnalysis.Rd index d26a2b4..3b6d4d0 100644 --- a/man/NetAnalysis.Rd +++ b/man/NetAnalysis.Rd @@ -4,7 +4,7 @@ \alias{NetAnalysis} \title{Networking analysis.} \usage{ -NetAnalysis(data, export = TRUE, FileName = "1") +NetAnalysis(data, export = TRUE, FileName = "NetworkAnalysisTable-1") } \arguments{ \item{data}{Protein-protein interaction data frame resulted from running the diff --git a/man/NoiseFiltering.Rd b/man/NoiseFiltering.Rd index d43d2c9..44f951e 100644 --- a/man/NoiseFiltering.Rd +++ b/man/NoiseFiltering.Rd @@ -15,7 +15,8 @@ NoiseFiltering( Val = TRUE, plot = TRUE, export = TRUE, - quiet = FALSE + quiet = FALSE, + filename = "Noise_filtering_genes_test" ) \S4method{NoiseFiltering}{DISCBIO}( @@ -28,7 +29,8 @@ NoiseFiltering( Val = TRUE, plot = TRUE, export = TRUE, - quiet = FALSE + quiet = FALSE, + filename = "Noise_filtering_genes_test" ) } \arguments{ @@ -56,6 +58,8 @@ Default is TRUE.} excel file. Default is TRUE.} \item{quiet}{if `TRUE`, suppresses printed output} + +\item{filename}{Name of the exported file (if `export=TRUE`)} } \value{ The DISCBIO-class object input with the noiseF slot filled. From ff725eb1357b8042b6db9ef6317d7dda92746e55 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Thu, 2 Jul 2020 12:25:49 +0200 Subject: [PATCH 33/99] Changed the default of export to FALSE --- R/DIscBIO-generic-DEGanalysis.R | 4 ++-- R/DIscBIO-generic-DEGanalysis2clust.R | 2 +- R/DIscBIO-generic-FinalPreprocessing.R | 4 ++-- R/DIscBIO-generic-KMClustDiffGenes.R | 2 +- R/DIscBIO-generic-KmeanOrder.R | 2 +- R/DIscBIO-generic-MBClustDiffGenes.R | 2 +- R/DIscBIO-generic-NoiseFiltering.R | 2 +- R/MB_Order.R | 2 +- R/NetAnalysis.R | 2 +- man/DEGanalysis.Rd | 4 ++-- man/DEGanalysis2clust.Rd | 4 ++-- man/FinalPreprocessing.Rd | 4 ++-- man/KMClustDiffGenes.Rd | 4 ++-- man/KmeanOrder.Rd | 4 ++-- man/MBClustDiffGenes.Rd | 4 ++-- man/MB_Order.Rd | 2 +- man/NetAnalysis.Rd | 2 +- man/NoiseFiltering.Rd | 4 ++-- 18 files changed, 27 insertions(+), 27 deletions(-) diff --git a/R/DIscBIO-generic-DEGanalysis.R b/R/DIscBIO-generic-DEGanalysis.R index b663909..46fbf63 100644 --- a/R/DIscBIO-generic-DEGanalysis.R +++ b/R/DIscBIO-generic-DEGanalysis.R @@ -36,7 +36,7 @@ setGeneric( Clustering = "K-means", fdr = 0.05, name = "Name", - export = TRUE, + export = FALSE, quiet = FALSE, plot = TRUE, filename_deg = "DEGsTable", @@ -57,7 +57,7 @@ setMethod( Clustering = "K-means", fdr = 0.05, name = "Name", - export = TRUE, + export = FALSE, quiet = FALSE, plot = TRUE, filename_deg, diff --git a/R/DIscBIO-generic-DEGanalysis2clust.R b/R/DIscBIO-generic-DEGanalysis2clust.R index beccd9c..66ca117 100644 --- a/R/DIscBIO-generic-DEGanalysis2clust.R +++ b/R/DIscBIO-generic-DEGanalysis2clust.R @@ -36,7 +36,7 @@ setGeneric( "DEGanalysis2clust", function( object, K, Clustering = "K-means", fdr = 0.05, name = "Name", - First = "CL1", Second = "CL2", export = TRUE, quiet = FALSE, + First = "CL1", Second = "CL2", export = FALSE, quiet = FALSE, plot = TRUE, filename_deg = "DEGsTable", filename_sigdeg = "sigDEG", ... ) diff --git a/R/DIscBIO-generic-FinalPreprocessing.R b/R/DIscBIO-generic-FinalPreprocessing.R index 996d505..2c00348 100644 --- a/R/DIscBIO-generic-FinalPreprocessing.R +++ b/R/DIscBIO-generic-FinalPreprocessing.R @@ -14,7 +14,7 @@ setGeneric( "FinalPreprocessing", function( - object, GeneFlitering = "NoiseF", export = TRUE, quiet = FALSE + object, GeneFlitering = "NoiseF", export = FALSE, quiet = FALSE ) standardGeneric("FinalPreprocessing") ) @@ -25,7 +25,7 @@ setMethod( "FinalPreprocessing", signature = "DISCBIO", definition = function( - object, GeneFlitering, export = TRUE, quiet = FALSE + object, GeneFlitering, export, quiet = FALSE ) { if (GeneFlitering == "NoiseF") { diff --git a/R/DIscBIO-generic-KMClustDiffGenes.R b/R/DIscBIO-generic-KMClustDiffGenes.R index 03ee8c5..c54f2b0 100644 --- a/R/DIscBIO-generic-KMClustDiffGenes.R +++ b/R/DIscBIO-generic-KMClustDiffGenes.R @@ -23,7 +23,7 @@ setGeneric( "KMClustDiffGenes", function( - object, K, pValue = 0.05, fdr = .01, export = TRUE, quiet = FALSE, + object, K, pValue = 0.05, fdr = .01, export = FALSE, quiet = FALSE, filename_up = "Up-DEG-cluster", filename_down = "Down-DEG-cluster", filename_binom = "binomial-DEGsTable", diff --git a/R/DIscBIO-generic-KmeanOrder.R b/R/DIscBIO-generic-KmeanOrder.R index a4061a0..b2747b2 100644 --- a/R/DIscBIO-generic-KmeanOrder.R +++ b/R/DIscBIO-generic-KmeanOrder.R @@ -13,7 +13,7 @@ #' sc <- valuesG1msReduced_treated_K #' Order <- KmeanOrder(sc, export = FALSE) #' Order@kordering -setGeneric("KmeanOrder", function(object, quiet = FALSE, export = TRUE, +setGeneric("KmeanOrder", function(object, quiet = FALSE, export = FALSE, filename = "Cellular_pseudo-time_ordering_based_on_k-meansc-lusters") standardGeneric("KmeanOrder") ) diff --git a/R/DIscBIO-generic-MBClustDiffGenes.R b/R/DIscBIO-generic-MBClustDiffGenes.R index 1ce620f..441a258 100644 --- a/R/DIscBIO-generic-MBClustDiffGenes.R +++ b/R/DIscBIO-generic-MBClustDiffGenes.R @@ -27,7 +27,7 @@ setGeneric( "MBClustDiffGenes", function( - object, K, pValue = 0.05, fdr = .01, export = TRUE, quiet = FALSE, + object, K, pValue = 0.05, fdr = .01, export = FALSE, quiet = FALSE, filename_up = "Up-DEG-cluster", filename_down = "Down-DEG-cluster", filename_binom = "binomial-DEGsTable", diff --git a/R/DIscBIO-generic-NoiseFiltering.R b/R/DIscBIO-generic-NoiseFiltering.R index e25046c..834f279 100644 --- a/R/DIscBIO-generic-NoiseFiltering.R +++ b/R/DIscBIO-generic-NoiseFiltering.R @@ -35,7 +35,7 @@ setGeneric( def = function( object, percentile = 0.8, CV = 0.3, geneCol = "yellow", FgeneCol = "black", erccCol = "blue", Val = TRUE, plot = TRUE, - export = TRUE, quiet = FALSE, + export = FALSE, quiet = FALSE, filename = "Noise_filtering_genes_test" ) standardGeneric("NoiseFiltering") diff --git a/R/MB_Order.R b/R/MB_Order.R index 96e51b1..4e9c273 100644 --- a/R/MB_Order.R +++ b/R/MB_Order.R @@ -17,7 +17,7 @@ #' sc@MBordering #' } MB_Order <- function( - object, quiet = FALSE, export = TRUE, + object, quiet = FALSE, export = FALSE, filename = "Cellular_pseudo-time_ordering_based_on_Model-based_clusters" ) { data = object@MBclusters diff --git a/R/NetAnalysis.R b/R/NetAnalysis.R index f024b82..218a08e 100644 --- a/R/NetAnalysis.R +++ b/R/NetAnalysis.R @@ -20,7 +20,7 @@ #' NetAnalysis(ppi, export=FALSE) #' } NetAnalysis <- function( - data, export = TRUE, FileName = "NetworkAnalysisTable-1" + data, export = FALSE, FileName = "NetworkAnalysisTable-1" ) { if (length(data[, 1]) < 1) stop("No Protein-Protein Interactions") diff --git a/man/DEGanalysis.Rd b/man/DEGanalysis.Rd index 0c1798f..64ce763 100644 --- a/man/DEGanalysis.Rd +++ b/man/DEGanalysis.Rd @@ -12,7 +12,7 @@ DEGanalysis( Clustering = "K-means", fdr = 0.05, name = "Name", - export = TRUE, + export = FALSE, quiet = FALSE, plot = TRUE, filename_deg = "DEGsTable", @@ -26,7 +26,7 @@ DEGanalysis( Clustering = "K-means", fdr = 0.05, name = "Name", - export = TRUE, + export = FALSE, quiet = FALSE, plot = TRUE, filename_deg = "DEGsTable", diff --git a/man/DEGanalysis2clust.Rd b/man/DEGanalysis2clust.Rd index 2bc3921..834d638 100644 --- a/man/DEGanalysis2clust.Rd +++ b/man/DEGanalysis2clust.Rd @@ -14,7 +14,7 @@ DEGanalysis2clust( name = "Name", First = "CL1", Second = "CL2", - export = TRUE, + export = FALSE, quiet = FALSE, plot = TRUE, filename_deg = "DEGsTable", @@ -30,7 +30,7 @@ DEGanalysis2clust( name = "Name", First = "CL1", Second = "CL2", - export = TRUE, + export = FALSE, quiet = FALSE, plot = TRUE, filename_deg = "DEGsTable", diff --git a/man/FinalPreprocessing.Rd b/man/FinalPreprocessing.Rd index feb0d3a..2a9f228 100644 --- a/man/FinalPreprocessing.Rd +++ b/man/FinalPreprocessing.Rd @@ -8,14 +8,14 @@ FinalPreprocessing( object, GeneFlitering = "NoiseF", - export = TRUE, + export = FALSE, quiet = FALSE ) \S4method{FinalPreprocessing}{DISCBIO}( object, GeneFlitering = "NoiseF", - export = TRUE, + export = FALSE, quiet = FALSE ) } diff --git a/man/KMClustDiffGenes.Rd b/man/KMClustDiffGenes.Rd index e6fd028..39dea6b 100644 --- a/man/KMClustDiffGenes.Rd +++ b/man/KMClustDiffGenes.Rd @@ -10,7 +10,7 @@ KMClustDiffGenes( K, pValue = 0.05, fdr = 0.01, - export = TRUE, + export = FALSE, quiet = FALSE, filename_up = "Up-DEG-cluster", filename_down = "Down-DEG-cluster", @@ -23,7 +23,7 @@ KMClustDiffGenes( K, pValue = 0.05, fdr = 0.01, - export = TRUE, + export = FALSE, quiet = FALSE, filename_up = "Up-DEG-cluster", filename_down = "Down-DEG-cluster", diff --git a/man/KmeanOrder.Rd b/man/KmeanOrder.Rd index 056891c..514c02c 100644 --- a/man/KmeanOrder.Rd +++ b/man/KmeanOrder.Rd @@ -8,14 +8,14 @@ KmeanOrder( object, quiet = FALSE, - export = TRUE, + export = FALSE, filename = "Cellular_pseudo-time_ordering_based_on_k-meansc-lusters" ) \S4method{KmeanOrder}{DISCBIO}( object, quiet = FALSE, - export = TRUE, + export = FALSE, filename = "Cellular_pseudo-time_ordering_based_on_k-meansc-lusters" ) } diff --git a/man/MBClustDiffGenes.Rd b/man/MBClustDiffGenes.Rd index 491911a..4d4d780 100644 --- a/man/MBClustDiffGenes.Rd +++ b/man/MBClustDiffGenes.Rd @@ -10,7 +10,7 @@ MBClustDiffGenes( K, pValue = 0.05, fdr = 0.01, - export = TRUE, + export = FALSE, quiet = FALSE, filename_up = "Up-DEG-cluster", filename_down = "Down-DEG-cluster", @@ -23,7 +23,7 @@ MBClustDiffGenes( K, pValue = 0.05, fdr = 0.01, - export = TRUE, + export = FALSE, quiet = FALSE, filename_up = "Up-DEG-cluster", filename_down = "Down-DEG-cluster", diff --git a/man/MB_Order.Rd b/man/MB_Order.Rd index b6f3e1f..f911588 100644 --- a/man/MB_Order.Rd +++ b/man/MB_Order.Rd @@ -7,7 +7,7 @@ MB_Order( object, quiet = FALSE, - export = TRUE, + export = FALSE, filename = "Cellular_pseudo-time_ordering_based_on_Model-based_clusters" ) } diff --git a/man/NetAnalysis.Rd b/man/NetAnalysis.Rd index 3b6d4d0..1dc1adb 100644 --- a/man/NetAnalysis.Rd +++ b/man/NetAnalysis.Rd @@ -4,7 +4,7 @@ \alias{NetAnalysis} \title{Networking analysis.} \usage{ -NetAnalysis(data, export = TRUE, FileName = "NetworkAnalysisTable-1") +NetAnalysis(data, export = FALSE, FileName = "NetworkAnalysisTable-1") } \arguments{ \item{data}{Protein-protein interaction data frame resulted from running the diff --git a/man/NoiseFiltering.Rd b/man/NoiseFiltering.Rd index 44f951e..17c34b6 100644 --- a/man/NoiseFiltering.Rd +++ b/man/NoiseFiltering.Rd @@ -14,7 +14,7 @@ NoiseFiltering( erccCol = "blue", Val = TRUE, plot = TRUE, - export = TRUE, + export = FALSE, quiet = FALSE, filename = "Noise_filtering_genes_test" ) @@ -28,7 +28,7 @@ NoiseFiltering( erccCol = "blue", Val = TRUE, plot = TRUE, - export = TRUE, + export = FALSE, quiet = FALSE, filename = "Noise_filtering_genes_test" ) From 9373eddf3abdfbafa898225bc29d95e86d9cf307 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Fri, 10 Jul 2020 09:14:36 +0200 Subject: [PATCH 34/99] Replaced copyright holder username --- LICENSE | 2 +- 1 file changed, 1 insertion(+), 1 deletion(-) diff --git a/LICENSE b/LICENSE index eaa36da..b02088b 100644 --- a/LICENSE +++ b/LICENSE @@ -1,2 +1,2 @@ YEAR: 2019 -COPYRIGHT HOLDER: SystemsBiologist \ No newline at end of file +COPYRIGHT HOLDER: Salim Ghannoum \ No newline at end of file From 0819e65a923cad4f803916296ffaf9c43c7f639e Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Fri, 10 Jul 2020 09:19:23 +0200 Subject: [PATCH 35/99] Remove example of internal function --- R/prepExampleDataset.R | 11 ++++++----- man/prepExampleDataset.Rd | 12 ++++++++---- 2 files changed, 14 insertions(+), 9 deletions(-) diff --git a/R/prepExampleDataset.R b/R/prepExampleDataset.R index 5641e9e..e834c58 100644 --- a/R/prepExampleDataset.R +++ b/R/prepExampleDataset.R @@ -3,11 +3,12 @@ #' several examples #' @param dataset Dataset used for transformation #' @param save save results? -#' @examples -#' \donttest{ -#' DIscBIO:::prepExampleDataset(valuesG1msReduced, save=FALSE) -#' } -#' @return rda file +#' @details This function serves the purpose of treating datasets such as +#' valuesG1msReduced to reduce examples of other functions by bypassing some +#' analysis steps covered in the vignettes. +#' @return Two rda files, ones for K-means clustering and another for +#' Model-based clustering. +#' @author Waldir Leoncio prepExampleDataset <- function(dataset, save=TRUE) { # ========================================================================== # Initial data treatment diff --git a/man/prepExampleDataset.Rd b/man/prepExampleDataset.Rd index 5b990b3..b3345f2 100644 --- a/man/prepExampleDataset.Rd +++ b/man/prepExampleDataset.Rd @@ -12,14 +12,18 @@ prepExampleDataset(dataset, save = TRUE) \item{save}{save results?} } \value{ -rda file +Two rda files, ones for K-means clustering and another for +Model-based clustering. } \description{ Internal function that prepares a pre-treated dataset for use in several examples } -\examples{ -\donttest{ - DIscBIO:::prepExampleDataset(valuesG1msReduced, save=FALSE) +\details{ +This function serves the purpose of treating datasets such as +valuesG1msReduced to reduce examples of other functions by bypassing some +analysis steps covered in the vignettes. } +\author{ +Waldir Leoncio } From 644be135aa84082048a1635e4024474bd2324c57 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Fri, 10 Jul 2020 09:23:17 +0200 Subject: [PATCH 36/99] Fixed wrong calling of on.exit after options --- R/DIscBIO-generic-Clustexp.R | 4 ++-- R/Networking.R | 2 +- 2 files changed, 3 insertions(+), 3 deletions(-) diff --git a/R/DIscBIO-generic-Clustexp.R b/R/DIscBIO-generic-Clustexp.R index f2bf8db..e716708 100644 --- a/R/DIscBIO-generic-Clustexp.R +++ b/R/DIscBIO-generic-Clustexp.R @@ -206,7 +206,7 @@ setMethod( NULL for (i in 1:runs) { opar <- options(show.error.messages = FALSE) - on.exit(par(opar)) + on.exit(options(opar)) repeat { kmm <- try(Kmeans( data, @@ -220,7 +220,7 @@ setMethod( break } opar <- options(show.error.messages = TRUE) - on.exit(par(opar)) + on.exit(options(opar)) swss <- sum(kmm$withinss) if (swss < minSS) { diff --git a/R/Networking.R b/R/Networking.R index bc9c686..0368430 100644 --- a/R/Networking.R +++ b/R/Networking.R @@ -71,7 +71,7 @@ Networking <- function( repr.plot.width = width_choice, repr.plot.height = height_choice ) - on.exit(par(opar)) + on.exit(options(opar)) } set_plot_dimensions(plot_width, plot_height) From 2acee7bcb00b5ebfcd7006fce1603af7944d7bf8 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Fri, 10 Jul 2020 10:11:36 +0200 Subject: [PATCH 37/99] Removed fixed seeds --- R/DIscBIO-generic-Clustexp.R | 82 ++++++++++++++---------------- R/DIscBIO-generic-Exprmclust.R | 46 ++++++++--------- R/DIscBIO-generic-FindOutliersKM.R | 3 +- R/DIscBIO-generic-MBclustheatmap.R | 12 ++--- R/DIscBIO-generic-Normalizedata.R | 6 +-- R/DIscBIO-generic-comptSNE.R | 6 +-- R/DIscBIO-generic-comptsneMB.R | 9 ++-- R/samr-adapted.R | 8 +-- man/Clustexp.Rd | 7 ++- man/MBclustheatmap.Rd | 18 ++++++- man/Normalizedata.Rd | 8 +-- man/comptSNE.Rd | 8 +-- man/comptsneMB.Rd | 11 ++-- tests/testthat/test.DEGfreeze.R | 4 +- tests/testthat/test.DIscBIO.IMP.R | 4 +- 15 files changed, 115 insertions(+), 117 deletions(-) diff --git a/R/DIscBIO-generic-Clustexp.R b/R/DIscBIO-generic-Clustexp.R index e716708..ac73cfa 100644 --- a/R/DIscBIO-generic-Clustexp.R +++ b/R/DIscBIO-generic-Clustexp.R @@ -23,8 +23,7 @@ #' statistics. Default is 50 #' @param cln Number of clusters to be used. Default is \code{NULL} and the #' cluster number is inferred by the saturation criterion. -#' @param rseed Integer number. Random seed to enforce reproducible clustering -#' results. Default is 17000. +#' @param rseed Random integer to enforce reproducible clustering results. #' @param quiet if `TRUE`, intermediate output is suppressed #' @importFrom stats as.dist cor kmeans #' @importFrom cluster clusGap maxSE @@ -39,7 +38,7 @@ #' sc <- Clustexp(sc, cln=3) setGeneric("Clustexp", function(object, clustnr = 20, bootnr = 50, metric = "pearson", do.gap = TRUE, SE.method = "Tibs2001SEmax", - SE.factor = .25, B.gap = 50, cln = 0, rseed = 17000, quiet = FALSE) + SE.factor = .25, B.gap = 50, cln = 0, rseed = NULL, quiet = FALSE) { standardGeneric("Clustexp") } @@ -51,7 +50,7 @@ setMethod( f = "Clustexp", signature = "DISCBIO", definition = function(object, clustnr, bootnr, metric, do.gap, SE.method, - SE.factor, B.gap, cln, rseed, quiet = FALSE) + SE.factor, B.gap, cln, rseed, quiet) { if (!is.numeric(clustnr)) stop("clustnr has to be a positive integer") @@ -97,8 +96,8 @@ setMethod( stop("cln has to be a non-negative integer") else if (round(cln) != cln | cln < 0) stop("cln has to be a non-negative integer") - if (!is.numeric(rseed)) - stop("rseed has to be numeric") + if (!is.null(rseed) & !is.numeric(rseed)) + stop("rseed has to be numeric or NULL") if (!do.gap & cln == 0) stop("cln has to be a positive integer or do.gap has to be TRUE") @@ -134,7 +133,7 @@ setMethod( SE.factor = .25, B.gap = 50, cln = 0, - rseed = 17000, + rseed = rseed, quiet = FALSE) { if (clustnr < 2) stop("Choose clustnr > 1") @@ -322,20 +321,19 @@ setMethod( out } - clb <- - clusterboot( - di, - B = bootnr, - distances = FALSE, - bootmethod = "boot", - clustermethod = KmeansCBI, - krange = cln, - scaling = FALSE, - multipleboot = FALSE, - bscompare = TRUE, - seed = rseed, - count = !quiet - ) + clb <- clusterboot( + di, + B = bootnr, + distances = FALSE, + bootmethod = "boot", + clustermethod = KmeansCBI, + krange = cln, + scaling = FALSE, + multipleboot = FALSE, + bscompare = TRUE, + seed = rseed, + count = !quiet + ) return(list( x = x, clb = clb, @@ -344,30 +342,26 @@ setMethod( )) } } - y <- - clustfun( - object@fdata, - clustnr, - bootnr, - metric, - do.gap, - SE.method, - SE.factor, - B.gap, - cln, - rseed, - quiet = quiet - ) - object@kmeans <- - list( - kpart = y$clb$result$partition, - jaccard = y$clb$bootmean, - gap = y$gpr - ) + y <- clustfun( + object@fdata, + clustnr, + bootnr, + metric, + do.gap, + SE.method, + SE.factor, + B.gap, + cln, + rseed = rseed, + quiet = quiet + ) + object@kmeans <- list( + kpart = y$clb$result$partition, + jaccard = y$clb$bootmean, + gap = y$gpr + ) object@distances <- as.matrix(y$di) - set.seed(111111) # fixed seed to keep the same colors - object@fcol <- - sample(rainbow(max(y$clb$result$partition))) + object@fcol <- rainbow(max(y$clb$result$partition)) object@cpart <- object@kmeans$kpart return(object) } diff --git a/R/DIscBIO-generic-Exprmclust.R b/R/DIscBIO-generic-Exprmclust.R index d0df1d1..d372805 100644 --- a/R/DIscBIO-generic-Exprmclust.R +++ b/R/DIscBIO-generic-Exprmclust.R @@ -28,29 +28,28 @@ #' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) #' sc <- Exprmclust(sc,K = 2) #' print(sc@MBclusters) - setGeneric( - name = "Exprmclust", - def = function(object, +setGeneric( +name = "Exprmclust", +def = function(object, + K = 3, + modelNames = "VVV", + reduce = TRUE, + cluster = NULL, + quiet = FALSE) +standardGeneric("Exprmclust") +) + +#' @export +#' @rdname Exprmclust +setMethod( +f = "Exprmclust", +signature = "DISCBIO", +definition = function(object, K = 3, modelNames = "VVV", reduce = TRUE, cluster = NULL, - quiet = FALSE) - standardGeneric("Exprmclust") - ) - - #' @export - #' @rdname Exprmclust - setMethod( - f = "Exprmclust", - signature = "DISCBIO", - definition = function(object, - K = 3, - modelNames = "VVV", - reduce = TRUE, - cluster = NULL, - quiet = FALSE) { - set.seed(12345) + quiet = FALSE) { obj <- object@fdata if (reduce) { sdev <- prcomp(t(obj), scale = T)$sdev[1:20] @@ -105,11 +104,11 @@ object@MBclusters <- full_List return(object) } - ) +) - #' @export - #' @rdname Exprmclust - setMethod( +#' @export +#' @rdname Exprmclust +setMethod( f = "Exprmclust", signature = "data.frame", definition = function(object, @@ -118,7 +117,6 @@ reduce = TRUE, cluster = NULL, quiet = FALSE) { - set.seed(12345) obj <- object if (reduce) { sdev <- prcomp(t(obj), scale = T)$sdev[1:20] diff --git a/R/DIscBIO-generic-FindOutliersKM.R b/R/DIscBIO-generic-FindOutliersKM.R index c9bc7c6..e51db3c 100644 --- a/R/DIscBIO-generic-FindOutliersKM.R +++ b/R/DIscBIO-generic-FindOutliersKM.R @@ -228,8 +228,7 @@ setMethod( object@cpart <- cpart - set.seed(111111) - object@fcol <- sample(rainbow(max(cpart))) + object@fcol <- rainbow(max(cpart)) p <- object@kmeans$kpart[order(object@kmeans$kpart, decreasing = FALSE)] x <- object@out$cprobs[names(p)] diff --git a/R/DIscBIO-generic-MBclustheatmap.R b/R/DIscBIO-generic-MBclustheatmap.R index 47fb31c..824b362 100644 --- a/R/DIscBIO-generic-MBclustheatmap.R +++ b/R/DIscBIO-generic-MBclustheatmap.R @@ -9,6 +9,7 @@ #' "ward.D", "ward.D2", "single", "complete", "average". Default is "single". #' @param plot if `TRUE`, plots the heatmap; otherwise, just prints cclmo #' @param quiet if `TRUE`, intermediary output is suppressed +#' @param rseed Random integer to fix random results. #' @importFrom stats hclust as.dist cor kmeans #' @importFrom cluster clusGap maxSE #' @importFrom fpc clusterboot kmeansCBI @@ -23,10 +24,7 @@ #' } setGeneric( "MBclustheatmap", - function( - object, hmethod="single", plot=TRUE, quiet=FALSE - ) - { + function(object, hmethod="single", plot=TRUE, quiet=FALSE, rseed=NULL) { standardGeneric("MBclustheatmap") } ) @@ -36,7 +34,7 @@ setGeneric( setMethod( "MBclustheatmap", signature = "DISCBIO", - definition = function(object, hmethod, plot, quiet) { + definition = function(object, hmethod, plot, quiet, rseed) { x <- object@fdata object@clusterpar$metric <- "pearson" dist.gen <- @@ -57,7 +55,7 @@ setMethod( SE.factor = .25, B.gap = 50, cln = 0, - rseed = 17000, + rseed = rseed, quiet = FALSE) { if (clustnr < 2) stop("Choose clustnr > 1") @@ -122,7 +120,7 @@ setMethod( SE.factor = .25, B.gap = 50, cln = 0, - rseed = 17000, + rseed = rseed, quiet = quiet ) object@distances <- as.matrix(y$di) diff --git a/R/DIscBIO-generic-Normalizedata.R b/R/DIscBIO-generic-Normalizedata.R index 71e67e5..84b0fc6 100644 --- a/R/DIscBIO-generic-Normalizedata.R +++ b/R/DIscBIO-generic-Normalizedata.R @@ -19,8 +19,8 @@ #' downsampled versions of the transcript count data. Default is 1 which means #' that sampling noise should be comparable across cells. For high numbers of #' dsn the data will become similar to the median normalization. -#' @param rseed Integer number. Random seed to enforce reproducible clustering -#' results. Default is 17000. +#' @param rseed Random integer to enforce reproducible clustering. +#' results #' @include DIscBIO-classes.R #' @return The DISCBIO-class object input with the ndata and fdata slots filled. #' @examples @@ -36,7 +36,7 @@ setGeneric( "Normalizedata", function( object, mintotal = 1000, minexpr = 0, minnumber = 0, maxexpr = Inf, - downsample = FALSE, dsn = 1, rseed = 17000 + downsample = FALSE, dsn = 1, rseed = NULL ) standardGeneric("Normalizedata") ) diff --git a/R/DIscBIO-generic-comptSNE.R b/R/DIscBIO-generic-comptSNE.R index a80d982..f57e939 100644 --- a/R/DIscBIO-generic-comptSNE.R +++ b/R/DIscBIO-generic-comptSNE.R @@ -2,8 +2,8 @@ #' @description This function is used to compute the t-Distributed Stochastic #' Neighbor Embedding (t-SNE). #' @param object \code{DISCBIO} class object. -#' @param rseed Integer number. Random seed to to yield exactly reproducible -#' maps across different runs. Default is 15555. +#' @param rseed Random integer to to yield reproducible maps across different +#' runs #' @param max_iter maximum number of iterations to perform. #' @param epoch The number of iterations in between update messages. #' @param quiet if `TRUE`, suppresses intermediate output @@ -18,7 +18,7 @@ #' head(sc@tsne) setGeneric( name = "comptSNE", - def = function(object, rseed = 15555, max_iter = 5000, epoch = 500, + def = function(object, rseed = NULL, max_iter = 5000, epoch = 500, quiet = FALSE, ...) { standardGeneric("comptSNE") } diff --git a/R/DIscBIO-generic-comptsneMB.R b/R/DIscBIO-generic-comptsneMB.R index 1b5a93d..ea7db0e 100644 --- a/R/DIscBIO-generic-comptsneMB.R +++ b/R/DIscBIO-generic-comptsneMB.R @@ -2,12 +2,11 @@ #' @description This function is used to compute the t-Distributed Stochastic #' Neighbor Embedding (t-SNE). #' @param object \code{DISCBIO} class object. -#' @param rseed Integer number. Random seed to to yield exactly reproducible -#' maps across different runs. Default is 15555. #' @param max_iter maximum number of iterations to perform. #' @param epoch The number of iterations in between update messages. -#' @param quiet if `TRUE`, suppresses intermediate output -#' @param ... other parameters to be passed to `tsne::tsne` +#' @param quiet if `TRUE`, suppresses intermediate output. +#' @param rseed Random number to be set for reproducible results. +#' @param ... other parameters to be passed to `tsne::tsne`. #' @importFrom tsne tsne #' @importFrom stats as.dist cor #' @return The DISCBIO-class object input with the MBtsne slot filled. @@ -24,7 +23,7 @@ #' print(sc@MBtsne) setGeneric( name = "comptsneMB", - def = function(object, rseed = 15555, max_iter = 5000, epoch = 500, + def = function(object, rseed = NULL, max_iter = 5000, epoch = 500, quiet = FALSE, ...) { standardGeneric("comptsneMB") } diff --git a/R/samr-adapted.R b/R/samr-adapted.R index eebb5d1..03a0d62 100644 --- a/R/samr-adapted.R +++ b/R/samr-adapted.R @@ -54,12 +54,8 @@ sammy <- function (data, resp.type = c("Quantitative", "Two class unpaired", resp.type.arg = match.arg(resp.type) assay.type = match.arg(assay.type) xl.mode = match.arg(xl.mode) - if (!is.null(random.seed)) { - set.seed(random.seed) - } - if (is.null(nresamp.perm)) { - nresamp.perm = nresamp - } + set.seed(random.seed) + if (is.null(nresamp.perm)) nresamp.perm <- nresamp nresamp.perm = min(nresamp, nresamp.perm) if (xl.mode == "regular" | xl.mode == "firsttime") { x = NULL diff --git a/man/Clustexp.Rd b/man/Clustexp.Rd index 8d4ba02..7b5698d 100644 --- a/man/Clustexp.Rd +++ b/man/Clustexp.Rd @@ -16,7 +16,7 @@ Clustexp( SE.factor = 0.25, B.gap = 50, cln = 0, - rseed = 17000, + rseed = NULL, quiet = FALSE ) @@ -30,7 +30,7 @@ Clustexp( SE.factor = 0.25, B.gap = 50, cln = 0, - rseed = 17000, + rseed = NULL, quiet = FALSE ) } @@ -65,8 +65,7 @@ statistics. Default is 50} \item{cln}{Number of clusters to be used. Default is \code{NULL} and the cluster number is inferred by the saturation criterion.} -\item{rseed}{Integer number. Random seed to enforce reproducible clustering -results. Default is 17000.} +\item{rseed}{Random integer to enforce reproducible clustering results.} \item{quiet}{if `TRUE`, intermediate output is suppressed} } diff --git a/man/MBclustheatmap.Rd b/man/MBclustheatmap.Rd index 4cd5858..2ba4aa3 100644 --- a/man/MBclustheatmap.Rd +++ b/man/MBclustheatmap.Rd @@ -6,9 +6,21 @@ \title{Plotting the Model-based clusters in a heatmap representation of the cell-to-cell distances} \usage{ -MBclustheatmap(object, hmethod = "single", plot = TRUE, quiet = FALSE) +MBclustheatmap( + object, + hmethod = "single", + plot = TRUE, + quiet = FALSE, + rseed = NULL +) -\S4method{MBclustheatmap}{DISCBIO}(object, hmethod = "single", plot = TRUE, quiet = FALSE) +\S4method{MBclustheatmap}{DISCBIO}( + object, + hmethod = "single", + plot = TRUE, + quiet = FALSE, + rseed = NULL +) } \arguments{ \item{object}{\code{DISCBIO} class object.} @@ -20,6 +32,8 @@ from hierarchical clustering of the cluster medoids. This should be one of \item{plot}{if `TRUE`, plots the heatmap; otherwise, just prints cclmo} \item{quiet}{if `TRUE`, intermediary output is suppressed} + +\item{rseed}{Random integer to fix random results.} } \value{ Unless otherwise specified, a heatmap and a vector of the underlying diff --git a/man/Normalizedata.Rd b/man/Normalizedata.Rd index 07c8c85..bda1241 100644 --- a/man/Normalizedata.Rd +++ b/man/Normalizedata.Rd @@ -13,7 +13,7 @@ Normalizedata( maxexpr = Inf, downsample = FALSE, dsn = 1, - rseed = 17000 + rseed = NULL ) \S4method{Normalizedata}{DISCBIO}( @@ -24,7 +24,7 @@ Normalizedata( maxexpr = Inf, downsample = FALSE, dsn = 1, - rseed = 17000 + rseed = NULL ) } \arguments{ @@ -53,8 +53,8 @@ downsampled versions of the transcript count data. Default is 1 which means that sampling noise should be comparable across cells. For high numbers of dsn the data will become similar to the median normalization.} -\item{rseed}{Integer number. Random seed to enforce reproducible clustering -results. Default is 17000.} +\item{rseed}{Random integer to enforce reproducible clustering. +results} } \value{ The DISCBIO-class object input with the ndata and fdata slots filled. diff --git a/man/comptSNE.Rd b/man/comptSNE.Rd index c3e56b3..73b784c 100644 --- a/man/comptSNE.Rd +++ b/man/comptSNE.Rd @@ -7,7 +7,7 @@ \usage{ comptSNE( object, - rseed = 15555, + rseed = NULL, max_iter = 5000, epoch = 500, quiet = FALSE, @@ -16,7 +16,7 @@ comptSNE( \S4method{comptSNE}{DISCBIO}( object, - rseed = 15555, + rseed = NULL, max_iter = 5000, epoch = 500, quiet = FALSE, @@ -26,8 +26,8 @@ comptSNE( \arguments{ \item{object}{\code{DISCBIO} class object.} -\item{rseed}{Integer number. Random seed to to yield exactly reproducible -maps across different runs. Default is 15555.} +\item{rseed}{Random integer to to yield reproducible maps across different +runs} \item{max_iter}{maximum number of iterations to perform.} diff --git a/man/comptsneMB.Rd b/man/comptsneMB.Rd index bcc8ba5..69f88eb 100644 --- a/man/comptsneMB.Rd +++ b/man/comptsneMB.Rd @@ -7,7 +7,7 @@ \usage{ comptsneMB( object, - rseed = 15555, + rseed = NULL, max_iter = 5000, epoch = 500, quiet = FALSE, @@ -16,7 +16,7 @@ comptsneMB( \S4method{comptsneMB}{DISCBIO}( object, - rseed = 15555, + rseed = NULL, max_iter = 5000, epoch = 500, quiet = FALSE, @@ -26,16 +26,15 @@ comptsneMB( \arguments{ \item{object}{\code{DISCBIO} class object.} -\item{rseed}{Integer number. Random seed to to yield exactly reproducible -maps across different runs. Default is 15555.} +\item{rseed}{Random number to be set for reproducible results.} \item{max_iter}{maximum number of iterations to perform.} \item{epoch}{The number of iterations in between update messages.} -\item{quiet}{if `TRUE`, suppresses intermediate output} +\item{quiet}{if `TRUE`, suppresses intermediate output.} -\item{...}{other parameters to be passed to `tsne::tsne`} +\item{...}{other parameters to be passed to `tsne::tsne`.} } \value{ The DISCBIO-class object input with the MBtsne slot filled. diff --git a/tests/testthat/test.DEGfreeze.R b/tests/testthat/test.DEGfreeze.R index 7d0c1ce..1948ce9 100644 --- a/tests/testthat/test.DEGfreeze.R +++ b/tests/testthat/test.DEGfreeze.R @@ -25,7 +25,9 @@ sc <- FinalPreprocessing(sc, GeneFlitering="ExpF", export=FALSE, quiet=TRUE) # ------------------------------------------------------------------------------ # DEG Analysis # ------------------------------------------------------------------------------ -sc <- Clustexp(sc, clustnr=n_clust, quiet=TRUE, bootnr = 2, B.gap = 2) +sc <- Clustexp( + sc, clustnr=n_clust, quiet=TRUE, bootnr = 2, B.gap = 2, rseed=17000 +) cdiff <- DEGanalysis( sc, Clustering="K-means", K=n_clust, fdr=0.10, name="all_clusters", export=FALSE, quiet=TRUE, plot=FALSE, nresamp=2, nperms=2 diff --git a/tests/testthat/test.DIscBIO.IMP.R b/tests/testthat/test.DIscBIO.IMP.R index b5e3925..643e2f7 100644 --- a/tests/testthat/test.DIscBIO.IMP.R +++ b/tests/testthat/test.DIscBIO.IMP.R @@ -46,7 +46,7 @@ test_that("Data is normalized", { context("K-means clustering") -sc <- Clustexp(sc, cln=2, quiet=TRUE) # K-means clustering +sc <- Clustexp(sc, cln=2, quiet=TRUE, rseed=17000) # K-means clustering sc <- comptSNE(sc, rseed=15555, quiet=TRUE, max_iter=10) test_that("tSNE is computed", { @@ -184,7 +184,7 @@ test_that("tSNE clustering works fine", { context("MB outliers") -sc <- Clustexp(sc, cln=2, quiet=TRUE) +sc <- Clustexp(sc, cln=2, quiet=TRUE, rseed=17000) Outliers <- FindOutliersMB( sc, K=2, outminc=5, outlg=2, probthr=.5*1e-3, thr=2 ^ (-1:-40), From 287d83f6e2492012763a22b07a36e2ed73a89186 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Fri, 10 Jul 2020 10:25:54 +0200 Subject: [PATCH 38/99] Minor simplification of vignette --- vignettes/DIscBIO-vignette.Rmd | 20 ++++++++++---------- 1 file changed, 10 insertions(+), 10 deletions(-) diff --git a/vignettes/DIscBIO-vignette.Rmd b/vignettes/DIscBIO-vignette.Rmd index 5ec169f..8357d53 100644 --- a/vignettes/DIscBIO-vignette.Rmd +++ b/vignettes/DIscBIO-vignette.Rmd @@ -9,7 +9,7 @@ vignette: > %\VignetteEncoding{UTF-8} --- -![DIscBIO](DiscBIO.png){width=90%} +![DIscBIO](DiscBIO.png){width=80%} The pipeline consists of four successive steps: data pre-processing, cellular clustering and pseudo-temporal ordering, determining differential expressed genes and identifying biomarkers. @@ -21,7 +21,7 @@ knitr::opts_chunk$set(fig.width=7, fig.height=7, eval=FALSE) ```{r} library(DIscBIO) -library(enrichR, quiet=TRUE) +library(enrichR) ``` ## Loading dataset @@ -46,7 +46,7 @@ The key idea in filtering genes is to appoint the genes that manifest abundant v In case the raw data does not include ERCC spike-ins, genes will be only filtered based on minimum expression in certain number of cells. -![DataPre-processing](Data-Preprocessing.png){width=90%} +![DataPre-processing](Data-Preprocessing.png){width=80%} ```{r} sc<- DISCBIO(DataSet) @@ -119,7 +119,7 @@ if (OnlyExpressionFiltering==TRUE){ Cellular clustering is performed according to the gene expression profiles to detect cellular sub-population with unique properties. After clustering, pseudo-temporal ordering is generated to indicate the cellular differentiation degree. -![PSCAN](Clustering.png){width=90%} +![PSCAN](Clustering.png){width=80%} ## 2.1. K-means Clustering @@ -140,7 +140,7 @@ The Clustexp() functions has several parameters: - rseed Integer number. Random seed to enforce reproducible clustering results. Default is 17000. - quiet if `TRUE`, intermediate output is suppressed -![DIsccBIO](KM1.png){width=90%} +![DIsccBIO](KM1.png){width=80%} ```{r} @@ -236,7 +236,7 @@ plotExptSNE(sc,g) Differentially expressed genes between individual clusters are identified using the significance analysis of sequencing data (SAMseq), which is a new function in significance analysis of microarrays (Li and Tibshirani 2011) in the samr package v2.0 (Tibshirani et all., 2015). SAMseq is a non-parametric statistical function dependent on Wilcoxon rank statistic that equalizes the sizes of the library by a resampling method accounting for the various sequencing depths. The analysis is implemented over the pure raw dataset that has the unnormalized expression read counts after excluding the ERCCs. Furthermore, DEGs in each cluster comparing to all the remaining clusters are determined using binomial differential expression, which is based on binomial counting statistics. -![DIsccBIO](KM2.png){width=90%} +![DIsccBIO](KM2.png){width=80%} ## 3.1 Identifying DEGs using SAMseq @@ -304,7 +304,7 @@ In case the user would like to get the names of the significant DEGs, then FS sh There are several methods to identify biomarkers, among them are decision trees and hub detection through networking analysis. The outcome of STRING analysis is stored in tab separated values (TSV) files. These TSV files served as an input to check both the connectivity degree and the betweenness centrality, which reflects the communication flow in the defined PPI networks -![DIsccBIO](KM3.png){width=90%} +![DIsccBIO](KM3.png){width=80%} Decision trees are one of the most efficient classification techniques in biomarkers discovery. Here we use it to predict the sub-population of a target cell based on transcriptomic data. Two types of decision trees can be performed: classification and regression trees (CART) and J48. The decision tree analysis is implemented over a training dataset, which consisted of the DEGs obtained by either SAMseq or the binomial differential expression. The performance of the generated trees can be evaluated for error estimation by ten-fold cross validation assessment using the "J48DTeval" and "RpartEVAL" functions. The decision tree analysis requires the dataset to be class vectored by applying the “ClassVectoringDT” function. @@ -418,7 +418,7 @@ GEA ## 2.2. Model-Based clustering -![DIsccBIO](MB1.png){width=90%} +![DIsccBIO](MB1.png){width=80%} Model-based clustering assumes that the data are generated by a model and attempts to recover the original model from the data to define cellular clusters. The steps reproduced here are comparable to the ones on Section [2.1](#21-k-means-clustering), and the output will be suppressed for brevity. @@ -532,7 +532,7 @@ PlotMBexpPCA(sc,g) Differentially expressed genes between individual clusters are identified using the significance analysis of sequencing data (SAMseq), which is a new function in significance analysis of microarrays (Li and Tibshirani 2011) in the samr package v2.0 (Tibshirani et all., 2015). SAMseq is a non-parametric statistical function dependent on Wilcoxon rank statistic that equalizes the sizes of the library by a resampling method accounting for the various sequencing depths. The analysis is implemented over the pure raw dataset that has the unnormalized expression read counts after excluding the ERCCs. Furthermore, DEGs in each cluster comparing to all the remaining clusters are determined using binomial differential expression, which is based on binomial counting statistics. -![DIsccBIO](MB2.png){width=90%} +![DIsccBIO](MB2.png){width=80%} ## 3.1 Identifying DEGs using SAMseq @@ -600,7 +600,7 @@ Vplot<-VolcanoPlot(U,value=0.001,name=name,FS=.5,fc=0.5) There are several methods to identify biomarkers, among them are decision trees and hub detection through networking analysis. The outcome of STRING analysis is stored in tab separated values (TSV) files. These TSV files served as an input to check both the connectivity degree and the betweenness centrality, which reflects the communication flow in the defined PPI networks -![DIsccBIO](MB3.png){width=90%} +![DIsccBIO](MB3.png){width=80%} Decision trees are one of the most efficient classification techniques in biomarkers discovery. Here we use it to predict the sub-population of a target cell based on transcriptomic data. Two types of decision trees can be performed: classification and regression trees (CART) and J48. The decision tree analysis is implemented over a training dataset, which consisted of the DEGs obtained by either SAMseq or the binomial differential expression. The performance of the generated trees can be evaluated for error estimation by ten-fold cross validation assessment using the "J48DTeval" and "RpartEVAL" functions. The decision tree analysis requires the dataset to be class vectored by applying the “ClassVectoringDT” function. From 7f4d48bcd68c4ce778e83f01ae005091172520ee Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Fri, 10 Jul 2020 10:45:41 +0200 Subject: [PATCH 39/99] Removed treated datasets and examples using them --- .Rbuildignore | 2 ++ R/DIscBIO-generic-ClassVectoringDT.R | 11 ----------- R/DIscBIO-generic-DEGanalysis.R | 7 ------- R/DIscBIO-generic-DEGanalysis2clust.R | 6 ------ R/DIscBIO-generic-KMclustheatmap.R | 4 ---- R/DIscBIO-generic-KmeanOrder.R | 5 ----- R/DIscBIO-generic-MBClustDiffGenes.R | 8 -------- R/DIscBIO-generic-MBclustheatmap.R | 7 ------- R/DIscBIO-generic-PlotmclustMB.R | 4 ---- R/DIscBIO-generic-plotExptSNE.R | 5 ----- R/DIscBIO-generic-plotGap.R | 4 ---- R/DIscBIO-generic-plotKmeansLabelstSNE.R | 4 ---- R/DIscBIO-generic-plotMBLabelstSNE.R | 4 ---- R/DIscBIO-generic-plotOrderKMtsne.R | 5 ----- R/DIscBIO-generic-plotOrderMBtsne.R | 4 ---- R/DIscBIO-generic-plotSilhouette.R | 4 ---- R/DIscBIO-generic-plotSymbolstSNE.R | 4 ---- R/DIscBIO-generic-plotexptsneMB.R | 8 -------- R/DIscBIO-generic-plotsilhouetteMB.R | 4 ---- R/DIscBIO-generic-plottSNE.R | 4 ---- R/DIscBIO-generic-plottsneMB.R | 4 ---- R/J48DT.R | 13 ------------- R/J48DTeval.R | 13 ------------- R/Jaccard.R | 4 ---- R/MB_Order.R | 7 ------- R/NetAnalysis.R | 8 -------- R/Networking.R | 7 ------- R/PPI.R | 7 ------- R/PlotMBexpPCA.R | 5 ----- R/PlotMBorderPCA.R | 6 ------ R/RpartDT.R | 12 ------------ R/RpartEVAL.R | 13 ------------- R/VolcanoPlot.R | 7 ------- man/ClassVectoringDT.Rd | 12 ------------ man/DEGanalysis.Rd | 8 -------- man/DEGanalysis2clust.Rd | 7 ------- man/J48DT.Rd | 14 -------------- man/J48DTeval.Rd | 13 ------------- man/Jaccard.Rd | 5 ----- man/KMclustheatmap.Rd | 5 ----- man/KmeanOrder.Rd | 6 ------ man/MBClustDiffGenes.Rd | 9 --------- man/MB_Order.Rd | 8 -------- man/MBclustheatmap.Rd | 8 -------- man/NetAnalysis.Rd | 9 --------- man/Networking.Rd | 8 -------- man/PPI.Rd | 8 -------- man/PlotMBexpPCA.Rd | 6 ------ man/PlotMBorderPCA.Rd | 6 ------ man/PlotmclustMB.Rd | 5 ----- man/RpartDT.Rd | 13 ------------- man/RpartEVAL.Rd | 13 ------------- man/VolcanoPlot.Rd | 8 -------- man/plotExptSNE.Rd | 6 ------ man/plotGap.Rd | 5 ----- man/plotKmeansLabelstSNE.Rd | 5 ----- man/plotMBLabelstSNE.Rd | 5 ----- man/plotOrderKMtsne.Rd | 6 ------ man/plotOrderMBtsne.Rd | 5 ----- man/plotSilhouette.Rd | 5 ----- man/plotSymbolstSNE.Rd | 5 ----- man/plotexptsneMB.Rd | 9 --------- man/plotsilhouetteMB.Rd | 5 ----- man/plottSNE.Rd | 5 ----- man/plottsneMB.Rd | 5 ----- 65 files changed, 2 insertions(+), 445 deletions(-) diff --git a/.Rbuildignore b/.Rbuildignore index 25ff9f8..c02729d 100644 --- a/.Rbuildignore +++ b/.Rbuildignore @@ -9,6 +9,8 @@ notebook data/valuesG1ms.rda data/genes20k.rda data/pan_indrop_matrix_8000cells_18556genes.rda +data/valuesG1msReduced_treated_K.rda +data/valuesG1msReduced_treated_MB.rda apt.txt install.R postBuild diff --git a/R/DIscBIO-generic-ClassVectoringDT.R b/R/DIscBIO-generic-ClassVectoringDT.R index 147f4d2..6aab1a8 100644 --- a/R/DIscBIO-generic-ClassVectoringDT.R +++ b/R/DIscBIO-generic-ClassVectoringDT.R @@ -14,17 +14,6 @@ #' @param quiet If `TRUE`, suppresses intermediary output #' @importFrom biomaRt useDataset useMart getBM #' @return A data frame. -#' @examples -#' data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details -#' sc <- valuesG1msReduced_treated_K -#' cdiff <- DEGanalysis2clust( -#' sc, Clustering="K-means", K=2, fdr=.2, name="Name", First="CL1", -#' Second="CL2", export=FALSE -#' ) -#' DATAforDT <- ClassVectoringDT( -#' sc, Clustering="K-means", K=2, First="CL1", Second="CL2", cdiff[[1]] -#' ) -#' str(DATAforDT) setGeneric( "ClassVectoringDT", function( diff --git a/R/DIscBIO-generic-DEGanalysis.R b/R/DIscBIO-generic-DEGanalysis.R index 46fbf63..3199a5e 100644 --- a/R/DIscBIO-generic-DEGanalysis.R +++ b/R/DIscBIO-generic-DEGanalysis.R @@ -21,13 +21,6 @@ #' @importFrom utils write.csv capture.output #' @param ... additional parameters to be passed to samr() #' @return A list containing two tables. -#' @examples -#' data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details -#' sc <- valuesG1msReduced_treated_K -#' DEGanalysis( -#' sc, Clustering="K-means", K=3, fdr=0.1, name="Name", export = FALSE, -#' nresamp=5, nperms=20 -#' ) setGeneric( name = "DEGanalysis", def = function( diff --git a/R/DIscBIO-generic-DEGanalysis2clust.R b/R/DIscBIO-generic-DEGanalysis2clust.R index 66ca117..5af9271 100644 --- a/R/DIscBIO-generic-DEGanalysis2clust.R +++ b/R/DIscBIO-generic-DEGanalysis2clust.R @@ -26,12 +26,6 @@ #' @importFrom utils write.csv capture.output #' @importFrom AnnotationDbi keys #' @return A list containing two tables. -#' @examples -#' data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details -#' sc <- valuesG1msReduced_treated_K -#' DEGanalysis2clust( -#' sc, Clustering="K-means", K=3, fdr=0.1, name="Name", export = FALSE -#' ) setGeneric( "DEGanalysis2clust", function( diff --git a/R/DIscBIO-generic-KMclustheatmap.R b/R/DIscBIO-generic-KMclustheatmap.R index f398445..f47e8c8 100644 --- a/R/DIscBIO-generic-KMclustheatmap.R +++ b/R/DIscBIO-generic-KMclustheatmap.R @@ -11,10 +11,6 @@ #' @return Unless otherwise specified, a heatmap and a vector of the underlying #' cluster order. #' @importFrom stats hclust as.dist cor -#' @examples -#' data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details -#' sc <- valuesG1msReduced_treated_K -#' KMclustheatmap(sc, hmethod="single") setGeneric( "KMclustheatmap", function( diff --git a/R/DIscBIO-generic-KmeanOrder.R b/R/DIscBIO-generic-KmeanOrder.R index b2747b2..5e8df8b 100644 --- a/R/DIscBIO-generic-KmeanOrder.R +++ b/R/DIscBIO-generic-KmeanOrder.R @@ -8,11 +8,6 @@ #' @param filename Name of the exported file (if `export=TRUE`) #' @importFrom TSCAN TSCANorder #' @return The DISCBIO-class object input with the kordering slot filled. -#' @examples -#' data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details -#' sc <- valuesG1msReduced_treated_K -#' Order <- KmeanOrder(sc, export = FALSE) -#' Order@kordering setGeneric("KmeanOrder", function(object, quiet = FALSE, export = FALSE, filename = "Cellular_pseudo-time_ordering_based_on_k-meansc-lusters") standardGeneric("KmeanOrder") diff --git a/R/DIscBIO-generic-MBClustDiffGenes.R b/R/DIscBIO-generic-MBClustDiffGenes.R index 441a258..ebaf110 100644 --- a/R/DIscBIO-generic-MBClustDiffGenes.R +++ b/R/DIscBIO-generic-MBClustDiffGenes.R @@ -16,14 +16,6 @@ #' @import org.Hs.eg.db #' @export #' @return A list containing two tables. -#' @examples -#' \donttest{ -#' data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset -#' sc <- valuesG1msReduced_treated_MB -#' sc <- MB_Order(sc, export = FALSE) -#' cdiff <- MBClustDiffGenes(sc, K=3, fdr=.1, export=FALSE) -#' str(cdiff) -#' } setGeneric( "MBClustDiffGenes", function( diff --git a/R/DIscBIO-generic-MBclustheatmap.R b/R/DIscBIO-generic-MBclustheatmap.R index 824b362..2994907 100644 --- a/R/DIscBIO-generic-MBclustheatmap.R +++ b/R/DIscBIO-generic-MBclustheatmap.R @@ -15,13 +15,6 @@ #' @importFrom fpc clusterboot kmeansCBI #' @return Unless otherwise specified, a heatmap and a vector of the underlying #' cluster order. -#' @examples -#' \donttest{ -#' data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset -#' sc <- valuesG1msReduced_treated_MB -#' sc <- MB_Order(sc, export = FALSE) -#' MBclustheatmap(sc, hmethod="single") -#' } setGeneric( "MBclustheatmap", function(object, hmethod="single", plot=TRUE, quiet=FALSE, rseed=NULL) { diff --git a/R/DIscBIO-generic-PlotmclustMB.R b/R/DIscBIO-generic-PlotmclustMB.R index a7d5eda..d9732fd 100644 --- a/R/DIscBIO-generic-PlotmclustMB.R +++ b/R/DIscBIO-generic-PlotmclustMB.R @@ -6,10 +6,6 @@ #' element_line unit element_text element_rect #' @importFrom igraph get.edgelist degree get.shortest.paths #' @return A plot of the PCA. -#' @examples -#' data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset -#' sc <- valuesG1msReduced_treated_MB -#' PlotmclustMB(sc) setGeneric("PlotmclustMB", function(object) standardGeneric("PlotmclustMB")) diff --git a/R/DIscBIO-generic-plotExptSNE.R b/R/DIscBIO-generic-plotExptSNE.R index 89aba63..b67e418 100644 --- a/R/DIscBIO-generic-plotExptSNE.R +++ b/R/DIscBIO-generic-plotExptSNE.R @@ -9,11 +9,6 @@ #' @param n String of characters representing the title of the plot. Default is #' NULL and the first element of \code{g} is chosen. #' @return t-SNE plot for one particular gene -#' @examples -#' data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details -#' sc <- valuesG1msReduced_treated_K -#' g <- 'ENSG00000001460' -#' plotExptSNE(sc, g) setGeneric("plotExptSNE", function(object, g, n = NULL) standardGeneric("plotExptSNE") diff --git a/R/DIscBIO-generic-plotGap.R b/R/DIscBIO-generic-plotGap.R index 70ad424..f878e48 100644 --- a/R/DIscBIO-generic-plotGap.R +++ b/R/DIscBIO-generic-plotGap.R @@ -4,10 +4,6 @@ #' @param object \code{DISCBIO} class object. #' @param y_limits 2-length numeric vector with the limits for the gap plot #' @return A plot of the gap statistics -#' @examples -#' data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details -#' sc <- valuesG1msReduced_treated_K -#' plotGap(sc) setGeneric( "plotGap", function(object, y_limits = NULL) standardGeneric("plotGap") ) diff --git a/R/DIscBIO-generic-plotKmeansLabelstSNE.R b/R/DIscBIO-generic-plotKmeansLabelstSNE.R index 47d7fc6..a033449 100644 --- a/R/DIscBIO-generic-plotKmeansLabelstSNE.R +++ b/R/DIscBIO-generic-plotKmeansLabelstSNE.R @@ -4,10 +4,6 @@ #' @rdname plotKmeansLabelstSNE #' @importFrom graphics text #' @return Plot containing the ID of the cells in each cluster -#' @examples -#' data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details -#' sc <- valuesG1msReduced_treated_K -#' plotKmeansLabelstSNE(sc) # Plots the ID of the cells in each cluster setGeneric("plotKmeansLabelstSNE", function(object) standardGeneric("plotKmeansLabelstSNE")) diff --git a/R/DIscBIO-generic-plotMBLabelstSNE.R b/R/DIscBIO-generic-plotMBLabelstSNE.R index 838349b..71d52c3 100644 --- a/R/DIscBIO-generic-plotMBLabelstSNE.R +++ b/R/DIscBIO-generic-plotMBLabelstSNE.R @@ -3,10 +3,6 @@ #' @param object \code{DISCBIO} class object. #' @importFrom graphics text #' @return A plot of the `object@MBtsne` values -#' @examples -#' data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset -#' sc <- valuesG1msReduced_treated_MB -#' plotMBLabelstSNE(sc) setGeneric("plotMBLabelstSNE", function(object) standardGeneric("plotMBLabelstSNE") ) diff --git a/R/DIscBIO-generic-plotOrderKMtsne.R b/R/DIscBIO-generic-plotOrderKMtsne.R index 1716a4e..d1d5f72 100644 --- a/R/DIscBIO-generic-plotOrderKMtsne.R +++ b/R/DIscBIO-generic-plotOrderKMtsne.R @@ -4,11 +4,6 @@ #' ordering. #' @param object \code{DISCBIO} class object. #' @return A plot of the pseudo-time ordering. -#' @examples -#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -#' sc <- valuesG1msReduced_treated_K -#' sc <- KmeanOrder(sc, export = FALSE) -#' plotOrderKMtsne(sc) setGeneric("plotOrderKMtsne", function(object) standardGeneric("plotOrderKMtsne")) diff --git a/R/DIscBIO-generic-plotOrderMBtsne.R b/R/DIscBIO-generic-plotOrderMBtsne.R index 4a9aa0e..22920ca 100644 --- a/R/DIscBIO-generic-plotOrderMBtsne.R +++ b/R/DIscBIO-generic-plotOrderMBtsne.R @@ -4,10 +4,6 @@ #' ordering. #' @param object \code{DISCBIO} class object. #' @return A plot of the pseudo-time ordering. -#' @examples -#' data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset -#' sc <- valuesG1msReduced_treated_MB -#' plotOrderMBtsne(sc) setGeneric("plotOrderMBtsne", function(object) standardGeneric("plotOrderMBtsne")) diff --git a/R/DIscBIO-generic-plotSilhouette.R b/R/DIscBIO-generic-plotSilhouette.R index c1249ea..ba9be84 100644 --- a/R/DIscBIO-generic-plotSilhouette.R +++ b/R/DIscBIO-generic-plotSilhouette.R @@ -11,10 +11,6 @@ #' @importFrom stats as.dist cor #' @importFrom cluster silhouette #' @return A silhouette plot -#' @examples -#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -#' sc <- valuesG1msReduced_treated_K -#' plotSilhouette(sc, K=3) setGeneric( name = "plotSilhouette", def = function(object, K) standardGeneric("plotSilhouette") diff --git a/R/DIscBIO-generic-plotSymbolstSNE.R b/R/DIscBIO-generic-plotSymbolstSNE.R index adefae8..099b6d3 100644 --- a/R/DIscBIO-generic-plotSymbolstSNE.R +++ b/R/DIscBIO-generic-plotSymbolstSNE.R @@ -6,10 +6,6 @@ #' @param legloc A keyword from the list "bottomright", "bottom", "bottomleft", #' "left", "topleft", "top", "topright", "right" and "center". Default is #' "bottomright" -#' @examples -#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -#' sc <- valuesG1msReduced_treated_K -#' plotSymbolstSNE(sc,types=sub("(\\_\\d+)$","", names(sc@ndata))) setGeneric( "plotSymbolstSNE", function(object, types = NULL, legloc = "bottomright") { diff --git a/R/DIscBIO-generic-plotexptsneMB.R b/R/DIscBIO-generic-plotexptsneMB.R index e729561..6e239e7 100644 --- a/R/DIscBIO-generic-plotexptsneMB.R +++ b/R/DIscBIO-generic-plotexptsneMB.R @@ -10,14 +10,6 @@ #' @param n String of characters representing the title of the plot. Default is #' NULL and the first element of \code{g} is chosen. #' @return t-SNE plot for one particular gene -#' @examples -#' \donttest{ -#' data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset -#' sc <- valuesG1msReduced_treated_MB -#' sc <- MB_Order(sc, export = FALSE) -#' g <- 'ENSG00000001460' -#' plotexptsneMB(sc, g) -#' } setGeneric("plotexptsneMB", function(object, g, n = NULL) standardGeneric("plotexptsneMB")) diff --git a/R/DIscBIO-generic-plotsilhouetteMB.R b/R/DIscBIO-generic-plotsilhouetteMB.R index 52ad55f..84893ba 100644 --- a/R/DIscBIO-generic-plotsilhouetteMB.R +++ b/R/DIscBIO-generic-plotsilhouetteMB.R @@ -11,10 +11,6 @@ #' @importFrom stats as.dist cor #' @importFrom cluster silhouette #' @return A silhouette plot -#' @examples -#' data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset -#' sc <- valuesG1msReduced_treated_MB -#' plotsilhouetteMB(sc, K=2) setGeneric("plotsilhouetteMB", function(object, K) standardGeneric("plotsilhouetteMB")) diff --git a/R/DIscBIO-generic-plottSNE.R b/R/DIscBIO-generic-plottSNE.R index 15414d9..1fb9636 100644 --- a/R/DIscBIO-generic-plottSNE.R +++ b/R/DIscBIO-generic-plottSNE.R @@ -3,10 +3,6 @@ #' @param object \code{DISCBIO} class object. #' @importFrom graphics text #' @return A plot of t-SNEs. -#' @examples -#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -#' sc <- valuesG1msReduced_treated_K -#' plottSNE(sc) setGeneric("plottSNE", function(object) standardGeneric("plottSNE")) diff --git a/R/DIscBIO-generic-plottsneMB.R b/R/DIscBIO-generic-plottsneMB.R index 3341658..64452ed 100644 --- a/R/DIscBIO-generic-plottsneMB.R +++ b/R/DIscBIO-generic-plottsneMB.R @@ -4,10 +4,6 @@ #' @param K A numeric value of the number of clusters #' @importFrom graphics text #' @return A plot of t-SNEs. -#' @examples -#' data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset -#' sc <- valuesG1msReduced_treated_MB -#' plottsneMB(sc) setGeneric( name = "plottsneMB", def = function(object, K = length(table(object@MBclusters$clusterid))) { diff --git a/R/J48DT.R b/R/J48DT.R index f120e3a..21f6dcc 100644 --- a/R/J48DT.R +++ b/R/J48DT.R @@ -12,19 +12,6 @@ #' @importFrom grid gpar #' @return Information about the J48 model and, by default, a plot of the #' decision tree. -#' @examples -#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -#' sc <- valuesG1msReduced_treated_K -#' cdiff <- DEGanalysis2clust( -#' sc, Clustering="K-means", K=3, fdr=.2, name="Name", First="CL1", -#' Second="CL2", export=FALSE, -#' nresamp=5, nperms=20 -#' ) -#' sigDEG <- cdiff[[1]] -#' DATAforDT <- ClassVectoringDT( -#' sc, Clustering="K-means", K=3, First="CL1", Second="CL2", sigDEG, -#' ) -#' J48DT(DATAforDT) J48DT <- function(data, quiet = FALSE, plot = TRUE) { msg <- NULL if (!is.data.frame(data)) { diff --git a/R/J48DTeval.R b/R/J48DTeval.R index 26814f8..670cc5c 100644 --- a/R/J48DTeval.R +++ b/R/J48DTeval.R @@ -12,19 +12,6 @@ #' @param quiet If `TRUE`, suppresses intermediary output #' @importFrom stats predict #' @return Statistics about the J48 model -#' @examples -#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -#' sc <- valuesG1msReduced_treated_K -#' cdiff <- DEGanalysis2clust( -#' sc, Clustering="K-means", K=3, fdr=.2, name="Name", First="CL1", -#' Second="CL2", export=FALSE -#' ) -#' sigDEG <- cdiff[[1]] -#' DATAforDT <- ClassVectoringDT( -#' sc, Clustering="K-means", K=3, First="CL1", Second="CL2", sigDEG -#' ) -#' J48DTeval(DATAforDT, num.folds=10, First="CL1", Second="CL2") - J48DTeval <- function( data, num.folds = 10, First = "CL1", Second = "CL2", quiet = FALSE ) diff --git a/R/Jaccard.R b/R/Jaccard.R index 5c82279..1680d53 100644 --- a/R/Jaccard.R +++ b/R/Jaccard.R @@ -15,10 +15,6 @@ #' @importFrom boot boot #' @importFrom graphics barplot box #' @return A plot of the mean Jaccard similarity coefficient per cluster. -#' @examples -#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -#' sc <- valuesG1msReduced_treated_K -#' Jaccard(sc, Clustering="K-means", K=3) Jaccard <- function( object, Clustering = "K-means", diff --git a/R/MB_Order.R b/R/MB_Order.R index 4e9c273..c8de73e 100644 --- a/R/MB_Order.R +++ b/R/MB_Order.R @@ -9,13 +9,6 @@ #' @param filename Name of the exported file (if `export=TRUE`) #' @importFrom TSCAN TSCANorder #' @return The DISCBIO-class object input with the MBordering slot filled. -#' @examples -#' \donttest{ -#' data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset -#' sc <- valuesG1msReduced_treated_MB -#' sc <- MB_Order(sc, export = FALSE) -#' sc@MBordering -#' } MB_Order <- function( object, quiet = FALSE, export = FALSE, filename = "Cellular_pseudo-time_ordering_based_on_Model-based_clusters" diff --git a/R/NetAnalysis.R b/R/NetAnalysis.R index 218a08e..769d27c 100644 --- a/R/NetAnalysis.R +++ b/R/NetAnalysis.R @@ -11,14 +11,6 @@ #' average.path.length get.adjacency V E mean_distance betweenness #' @importFrom NetIndices GenInd #' @return A network analysis table -#' @examples -#' \donttest{ -#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -#' sc <- valuesG1msReduced_treated_K -#' dff <- DEGanalysis2clust(sc, K=3, export=FALSE, quiet=TRUE, plot=FALSE) -#' ppi <- PPI(dff$FinalDEGsU[, "Gene Name"]) -#' NetAnalysis(ppi, export=FALSE) -#' } NetAnalysis <- function( data, export = FALSE, FileName = "NetworkAnalysisTable-1" ) { diff --git a/R/Networking.R b/R/Networking.R index 0368430..5916625 100644 --- a/R/Networking.R +++ b/R/Networking.R @@ -13,13 +13,6 @@ #' @importFrom png readPNG #' @importFrom graphics plot rasterImage #' @return A plot of the network -#' @examples -#' \donttest{ -#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -#' sc <- valuesG1msReduced_treated_K -#' dff <- DEGanalysis2clust(sc, K=3, export=FALSE, quiet=TRUE, plot=FALSE) -#' Networking(dff$FinalDEGsU[, "Gene Name"]) -#' } Networking <- function( data, FileName = NULL, diff --git a/R/PPI.R b/R/PPI.R index 1e5eef1..cc22cef 100644 --- a/R/PPI.R +++ b/R/PPI.R @@ -11,13 +11,6 @@ #' @return Either a TSV file stored in the user's file system and its #' corresponding `data.frame` object in R or and R object containing that #' information. -#' @examples -#' \donttest{ -#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -#' sc <- valuesG1msReduced_treated_K -#' dff <- DEGanalysis2clust(sc, K=3, export=FALSE, quiet=TRUE, plot=FALSE) -#' PPI(dff$FinalDEGsU[, "Gene Name"]) -#' } PPI <- function(data, FileName = NULL, species = "9606") { # Save base enpoint as variable string_api_url <- "https://string-db.org/api/" diff --git a/R/PlotMBexpPCA.R b/R/PlotMBexpPCA.R index 30bf9d0..55eb822 100644 --- a/R/PlotMBexpPCA.R +++ b/R/PlotMBexpPCA.R @@ -11,11 +11,6 @@ #' @importFrom grDevices colorRampPalette #' @importFrom graphics layout par image #' @return A plot of the PCA. -#' @examples -#' data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset -#' sc <- valuesG1msReduced_treated_MB -#' g <- "ENSG00000010244" # Plotting the expression of MT-RNR2 -#' PlotMBexpPCA(sc, g) #' @export PlotMBexpPCA <- function(object, g, n = NULL) { diff --git a/R/PlotMBorderPCA.R b/R/PlotMBorderPCA.R index 58c6523..2d618f1 100644 --- a/R/PlotMBorderPCA.R +++ b/R/PlotMBorderPCA.R @@ -7,12 +7,6 @@ #' @importFrom graphics layout par image #' @export #' @return A plot of the PCA. -#' @examples -#' data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset -#' sc <- valuesG1msReduced_treated_MB -#' sc <- MB_Order(sc, export = FALSE) -#' PlotMBorderPCA(sc) - PlotMBorderPCA <- function(object) { data = object@MBclusters MBordertable <- cbind(data$pcareduceres, object@MBordering) diff --git a/R/RpartDT.R b/R/RpartDT.R index 89256d2..3a19521 100644 --- a/R/RpartDT.R +++ b/R/RpartDT.R @@ -10,18 +10,6 @@ #' @importFrom rpart.plot rpart.plot #' @return Information about the model and, by default, a plot of the decision #' tree. -#' @examples -#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -#' sc <- valuesG1msReduced_treated_K -#' cdiff <- DEGanalysis2clust( -#' sc, Clustering="K-means", K=3, fdr=.2, name="Name", First="CL1", -#' Second="CL2", export=FALSE -#' ) -#' sigDEG <- cdiff[[1]] -#' DATAforDT <- ClassVectoringDT( -#' sc, Clustering="K-means", K=3, First="CL1", Second="CL2", sigDEG, -#' ) -#' RpartDT(DATAforDT) RpartDT <- function(data, quiet = FALSE, plot = TRUE) { exp.df <- as.data.frame(t(data)) classVector <- factor(colnames(data)) diff --git a/R/RpartEVAL.R b/R/RpartEVAL.R index 724405e..ac11e3a 100644 --- a/R/RpartEVAL.R +++ b/R/RpartEVAL.R @@ -12,19 +12,6 @@ #' @param quiet If `TRUE`, suppresses intermediary output #' @importFrom stats predict #' @return Performance statistics of the model -#' @examples -#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -#' sc <- valuesG1msReduced_treated_K -#' cdiff <- DEGanalysis2clust( -#' sc, Clustering="K-means", K=3, fdr=.2, name="Name", First="CL1", -#' Second="CL2", export=FALSE -#' ) -#' sigDEG <- cdiff[[1]] -#' DATAforDT <- ClassVectoringDT( -#' sc, Clustering="K-means", K=3, First="CL1", Second="CL2", sigDEG, -#' ) -#' RpartEVAL(DATAforDT,num.folds=10,First="CL1",Second="CL2") - RpartEVAL <- function(data, num.folds = 10, First = "CL1", Second = "CL2", quiet = FALSE) { exp.imput.df <- as.data.frame(t(data)) diff --git a/R/VolcanoPlot.R b/R/VolcanoPlot.R index 9cb4549..37d017f 100644 --- a/R/VolcanoPlot.R +++ b/R/VolcanoPlot.R @@ -16,13 +16,6 @@ #' @importFrom calibrate textxy #' @return A volcano plot #' @export -#' @examples -#' \donttest{ -#' data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -#' sc <- valuesG1msReduced_treated_K -#' dff <- DEGanalysis2clust(sc, K=3, export=FALSE, quiet=TRUE, plot=FALSE) -#' VolcanoPlot(dff$FinalDEGsU, value=0.05, FS=.4) -#' } VolcanoPlot <- function(object, value = 0.05, name = NULL, fc = 0.5, FS = .4) { if (length(object[1, ]) > 8) { object <- object[, -1] diff --git a/man/ClassVectoringDT.Rd b/man/ClassVectoringDT.Rd index 013ff44..c6f1c38 100644 --- a/man/ClassVectoringDT.Rd +++ b/man/ClassVectoringDT.Rd @@ -51,15 +51,3 @@ A data frame. This function generates a class vector for the input dataset so the decision tree analysis can be implemented afterwards. } -\examples{ -data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details -sc <- valuesG1msReduced_treated_K -cdiff <- DEGanalysis2clust( - sc, Clustering="K-means", K=2, fdr=.2, name="Name", First="CL1", - Second="CL2", export=FALSE -) -DATAforDT <- ClassVectoringDT( - sc, Clustering="K-means", K=2, First="CL1", Second="CL2", cdiff[[1]] -) -str(DATAforDT) -} diff --git a/man/DEGanalysis.Rd b/man/DEGanalysis.Rd index 64ce763..caf4744 100644 --- a/man/DEGanalysis.Rd +++ b/man/DEGanalysis.Rd @@ -67,11 +67,3 @@ A list containing two tables. This function defines DEGs between all individual clusters generated by either K-means or model based clustering. } -\examples{ -data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details -sc <- valuesG1msReduced_treated_K -DEGanalysis( - sc, Clustering="K-means", K=3, fdr=0.1, name="Name", export = FALSE, - nresamp=5, nperms=20 -) -} diff --git a/man/DEGanalysis2clust.Rd b/man/DEGanalysis2clust.Rd index 834d638..7d2314a 100644 --- a/man/DEGanalysis2clust.Rd +++ b/man/DEGanalysis2clust.Rd @@ -77,10 +77,3 @@ A list containing two tables. This function defines DEGs between particular clusters generated by either K-means or model based clustering. } -\examples{ -data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details -sc <- valuesG1msReduced_treated_K -DEGanalysis2clust( - sc, Clustering="K-means", K=3, fdr=0.1, name="Name", export = FALSE -) -} diff --git a/man/J48DT.Rd b/man/J48DT.Rd index c5225e1..714a7d1 100644 --- a/man/J48DT.Rd +++ b/man/J48DT.Rd @@ -22,17 +22,3 @@ The decision tree analysis is implemented over a training dataset, which consisted of the DEGs obtained by either SAMseq or the binomial differential expression. } -\examples{ -data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -sc <- valuesG1msReduced_treated_K -cdiff <- DEGanalysis2clust( - sc, Clustering="K-means", K=3, fdr=.2, name="Name", First="CL1", - Second="CL2", export=FALSE, - nresamp=5, nperms=20 -) -sigDEG <- cdiff[[1]] -DATAforDT <- ClassVectoringDT( - sc, Clustering="K-means", K=3, First="CL1", Second="CL2", sigDEG, -) -J48DT(DATAforDT) -} diff --git a/man/J48DTeval.Rd b/man/J48DTeval.Rd index f6c648c..33c073d 100644 --- a/man/J48DTeval.Rd +++ b/man/J48DTeval.Rd @@ -27,16 +27,3 @@ Statistics about the J48 model This function evaluates the performance of the generated trees for error estimation by ten-fold cross validation assessment. } -\examples{ -data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -sc <- valuesG1msReduced_treated_K -cdiff <- DEGanalysis2clust( - sc, Clustering="K-means", K=3, fdr=.2, name="Name", First="CL1", - Second="CL2", export=FALSE -) -sigDEG <- cdiff[[1]] -DATAforDT <- ClassVectoringDT( - sc, Clustering="K-means", K=3, First="CL1", Second="CL2", sigDEG -) -J48DTeval(DATAforDT, num.folds=10, First="CL1", Second="CL2") -} diff --git a/man/Jaccard.Rd b/man/Jaccard.Rd index 2d5a0cd..8c14dcf 100644 --- a/man/Jaccard.Rd +++ b/man/Jaccard.Rd @@ -29,8 +29,3 @@ Robustness of the clusters can be assessed by Jaccard’s across bootstrapping runs. Jaccard’s similarity is the intersect of two clusters divided by the union. } -\examples{ -data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -sc <- valuesG1msReduced_treated_K -Jaccard(sc, Clustering="K-means", K=3) -} diff --git a/man/KMclustheatmap.Rd b/man/KMclustheatmap.Rd index 01e0f56..9a1c4c8 100644 --- a/man/KMclustheatmap.Rd +++ b/man/KMclustheatmap.Rd @@ -28,8 +28,3 @@ This functions plots a heatmap of the distance matrix grouped by clusters. Individual clusters are highlighted with rainbow colors along the x and y-axes. } -\examples{ -data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details -sc <- valuesG1msReduced_treated_K -KMclustheatmap(sc, hmethod="single") -} diff --git a/man/KmeanOrder.Rd b/man/KmeanOrder.Rd index 514c02c..4c97fbe 100644 --- a/man/KmeanOrder.Rd +++ b/man/KmeanOrder.Rd @@ -36,9 +36,3 @@ This function takes the exact output of exprmclust function and construct Pseudo-time ordering by mapping all cells onto the path that connects cluster centers. } -\examples{ -data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details -sc <- valuesG1msReduced_treated_K -Order <- KmeanOrder(sc, export = FALSE) -Order@kordering -} diff --git a/man/MBClustDiffGenes.Rd b/man/MBClustDiffGenes.Rd index 4d4d780..c4ebd26 100644 --- a/man/MBClustDiffGenes.Rd +++ b/man/MBClustDiffGenes.Rd @@ -59,12 +59,3 @@ A list containing two tables. \description{ ClustDiffGenes } -\examples{ -\donttest{ -data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset -sc <- valuesG1msReduced_treated_MB -sc <- MB_Order(sc, export = FALSE) -cdiff <- MBClustDiffGenes(sc, K=3, fdr=.1, export=FALSE) -str(cdiff) -} -} diff --git a/man/MB_Order.Rd b/man/MB_Order.Rd index f911588..70c2739 100644 --- a/man/MB_Order.Rd +++ b/man/MB_Order.Rd @@ -28,11 +28,3 @@ This function takes the exact output of exprmclust function and construct Pseudo-time ordering by mapping all cells onto the path that connects cluster centers. } -\examples{ -\donttest{ -data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset -sc <- valuesG1msReduced_treated_MB -sc <- MB_Order(sc, export = FALSE) -sc@MBordering -} -} diff --git a/man/MBclustheatmap.Rd b/man/MBclustheatmap.Rd index 2ba4aa3..9d993c4 100644 --- a/man/MBclustheatmap.Rd +++ b/man/MBclustheatmap.Rd @@ -44,11 +44,3 @@ This functions plots a heatmap of the distance matrix grouped by clusters. Individual clusters are highlighted with rainbow colors along the x and y-axes. } -\examples{ -\donttest{ -data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset -sc <- valuesG1msReduced_treated_MB -sc <- MB_Order(sc, export = FALSE) -MBclustheatmap(sc, hmethod="single") -} -} diff --git a/man/NetAnalysis.Rd b/man/NetAnalysis.Rd index 1dc1adb..cfa0006 100644 --- a/man/NetAnalysis.Rd +++ b/man/NetAnalysis.Rd @@ -22,12 +22,3 @@ This function checks the connectivity degree and the betweenness centrality, which reflect the communication flow in the defined PPI networks } -\examples{ -\donttest{ -data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -sc <- valuesG1msReduced_treated_K -dff <- DEGanalysis2clust(sc, K=3, export=FALSE, quiet=TRUE, plot=FALSE) -ppi <- PPI(dff$FinalDEGsU[, "Gene Name"]) -NetAnalysis(ppi, export=FALSE) -} -} diff --git a/man/Networking.Rd b/man/Networking.Rd index a12694a..6126ca8 100644 --- a/man/Networking.Rd +++ b/man/Networking.Rd @@ -31,11 +31,3 @@ A plot of the network \description{ This function uses STRING-api to plot the network. } -\examples{ -\donttest{ -data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -sc <- valuesG1msReduced_treated_K -dff <- DEGanalysis2clust(sc, K=3, export=FALSE, quiet=TRUE, plot=FALSE) -Networking(dff$FinalDEGsU[, "Gene Name"]) -} -} diff --git a/man/PPI.Rd b/man/PPI.Rd index 7b163d2..d52bae7 100644 --- a/man/PPI.Rd +++ b/man/PPI.Rd @@ -23,11 +23,3 @@ information. This function uses STRING-api. The outcome of STRING analysis will be stored in tab separated values (TSV) files. } -\examples{ -\donttest{ -data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -sc <- valuesG1msReduced_treated_K -dff <- DEGanalysis2clust(sc, K=3, export=FALSE, quiet=TRUE, plot=FALSE) -PPI(dff$FinalDEGsU[, "Gene Name"]) -} -} diff --git a/man/PlotMBexpPCA.Rd b/man/PlotMBexpPCA.Rd index 6b7e48c..f946b93 100644 --- a/man/PlotMBexpPCA.Rd +++ b/man/PlotMBexpPCA.Rd @@ -23,9 +23,3 @@ A plot of the PCA. The PCA representation can also be used to show the gene expression of a particular gene. } -\examples{ -data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset -sc <- valuesG1msReduced_treated_MB -g <- "ENSG00000010244" # Plotting the expression of MT-RNR2 -PlotMBexpPCA(sc, g) -} diff --git a/man/PlotMBorderPCA.Rd b/man/PlotMBorderPCA.Rd index 43c540a..cdc4775 100644 --- a/man/PlotMBorderPCA.Rd +++ b/man/PlotMBorderPCA.Rd @@ -16,9 +16,3 @@ A plot of the PCA. The PCA representation can also be used to show the pseudo-time ordering. } -\examples{ -data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset -sc <- valuesG1msReduced_treated_MB -sc <- MB_Order(sc, export = FALSE) -PlotMBorderPCA(sc) -} diff --git a/man/PlotmclustMB.Rd b/man/PlotmclustMB.Rd index c62a0bf..8262f83 100644 --- a/man/PlotmclustMB.Rd +++ b/man/PlotmclustMB.Rd @@ -18,8 +18,3 @@ A plot of the PCA. \description{ Plot the model-based clustering results } -\examples{ -data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset -sc <- valuesG1msReduced_treated_MB -PlotmclustMB(sc) -} diff --git a/man/RpartDT.Rd b/man/RpartDT.Rd index fa97410..e9eee75 100644 --- a/man/RpartDT.Rd +++ b/man/RpartDT.Rd @@ -22,16 +22,3 @@ The decision tree analysis is implemented over a training dataset, which consisted of the DEGs obtained by either SAMseq or the binomial differential expression. } -\examples{ -data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -sc <- valuesG1msReduced_treated_K -cdiff <- DEGanalysis2clust( - sc, Clustering="K-means", K=3, fdr=.2, name="Name", First="CL1", - Second="CL2", export=FALSE -) -sigDEG <- cdiff[[1]] -DATAforDT <- ClassVectoringDT( - sc, Clustering="K-means", K=3, First="CL1", Second="CL2", sigDEG, -) -RpartDT(DATAforDT) -} diff --git a/man/RpartEVAL.Rd b/man/RpartEVAL.Rd index 5fcd1e0..f594ae0 100644 --- a/man/RpartEVAL.Rd +++ b/man/RpartEVAL.Rd @@ -27,16 +27,3 @@ Performance statistics of the model This function evaluates the performance of the generated trees for error estimation by ten-fold cross validation assessment. } -\examples{ -data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -sc <- valuesG1msReduced_treated_K -cdiff <- DEGanalysis2clust( - sc, Clustering="K-means", K=3, fdr=.2, name="Name", First="CL1", - Second="CL2", export=FALSE -) -sigDEG <- cdiff[[1]] -DATAforDT <- ClassVectoringDT( - sc, Clustering="K-means", K=3, First="CL1", Second="CL2", sigDEG, -) -RpartEVAL(DATAforDT,num.folds=10,First="CL1",Second="CL2") -} diff --git a/man/VolcanoPlot.Rd b/man/VolcanoPlot.Rd index 37387ee..b464f89 100644 --- a/man/VolcanoPlot.Rd +++ b/man/VolcanoPlot.Rd @@ -27,11 +27,3 @@ Plotting differentially expressed genes (DEGs) in a particular cluster. Volcano plots are used to readily show the DEGs by plotting significance versus fold-change on the y and x axes, respectively. } -\examples{ -\donttest{ -data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -sc <- valuesG1msReduced_treated_K -dff <- DEGanalysis2clust(sc, K=3, export=FALSE, quiet=TRUE, plot=FALSE) -VolcanoPlot(dff$FinalDEGsU, value=0.05, FS=.4) -} -} diff --git a/man/plotExptSNE.Rd b/man/plotExptSNE.Rd index 1bcf8a3..56d250b 100644 --- a/man/plotExptSNE.Rd +++ b/man/plotExptSNE.Rd @@ -27,9 +27,3 @@ The t-SNE map representation can also be used to analyze expression of a gene or a group of genes, to investigate cluster specific gene expression patterns } -\examples{ -data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details -sc <- valuesG1msReduced_treated_K -g <- 'ENSG00000001460' -plotExptSNE(sc, g) -} diff --git a/man/plotGap.Rd b/man/plotGap.Rd index 6fd97e6..330b805 100644 --- a/man/plotGap.Rd +++ b/man/plotGap.Rd @@ -20,8 +20,3 @@ A plot of the gap statistics \description{ Plotting Gap Statistics } -\examples{ -data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details -sc <- valuesG1msReduced_treated_K -plotGap(sc) -} diff --git a/man/plotKmeansLabelstSNE.Rd b/man/plotKmeansLabelstSNE.Rd index 94153ee..c6185aa 100644 --- a/man/plotKmeansLabelstSNE.Rd +++ b/man/plotKmeansLabelstSNE.Rd @@ -18,8 +18,3 @@ Plot containing the ID of the cells in each cluster \description{ Visualizing the K-means clusters using tSNE maps } -\examples{ -data(valuesG1msReduced_treated_K) # DIscBIO:::prepExampleDataset for details -sc <- valuesG1msReduced_treated_K -plotKmeansLabelstSNE(sc) # Plots the ID of the cells in each cluster -} diff --git a/man/plotMBLabelstSNE.Rd b/man/plotMBLabelstSNE.Rd index 014d2c0..7b67822 100644 --- a/man/plotMBLabelstSNE.Rd +++ b/man/plotMBLabelstSNE.Rd @@ -18,8 +18,3 @@ A plot of the `object@MBtsne` values \description{ Visualizing the Model-based clusters using tSNE maps } -\examples{ -data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset -sc <- valuesG1msReduced_treated_MB -plotMBLabelstSNE(sc) -} diff --git a/man/plotOrderKMtsne.Rd b/man/plotOrderKMtsne.Rd index 81e1848..7d220eb 100644 --- a/man/plotOrderKMtsne.Rd +++ b/man/plotOrderKMtsne.Rd @@ -20,9 +20,3 @@ A plot of the pseudo-time ordering. The tSNE representation can also be used to show the pseudo-time ordering. } -\examples{ -data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -sc <- valuesG1msReduced_treated_K -sc <- KmeanOrder(sc, export = FALSE) -plotOrderKMtsne(sc) -} diff --git a/man/plotOrderMBtsne.Rd b/man/plotOrderMBtsne.Rd index bd93d9b..fdd099e 100644 --- a/man/plotOrderMBtsne.Rd +++ b/man/plotOrderMBtsne.Rd @@ -20,8 +20,3 @@ A plot of the pseudo-time ordering. The tSNE representation can also be used to show the pseudo-time ordering. } -\examples{ -data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset -sc <- valuesG1msReduced_treated_MB -plotOrderMBtsne(sc) -} diff --git a/man/plotSilhouette.Rd b/man/plotSilhouette.Rd index cea359c..074dcb4 100644 --- a/man/plotSilhouette.Rd +++ b/man/plotSilhouette.Rd @@ -30,8 +30,3 @@ The silhouette provides a representation of how well each point take values between -1 and 1 with higher values reflecting better representation of a point by its cluster. } -\examples{ -data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -sc <- valuesG1msReduced_treated_K -plotSilhouette(sc, K=3) -} diff --git a/man/plotSymbolstSNE.Rd b/man/plotSymbolstSNE.Rd index f4a4036..0152863 100644 --- a/man/plotSymbolstSNE.Rd +++ b/man/plotSymbolstSNE.Rd @@ -25,8 +25,3 @@ Plot of tsne objet slot, grouped by gene. \description{ Visualizing the K-means clusters using tSNE maps } -\examples{ -data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -sc <- valuesG1msReduced_treated_K -plotSymbolstSNE(sc,types=sub("(\\\\_\\\\d+)$","", names(sc@ndata))) -} diff --git a/man/plotexptsneMB.Rd b/man/plotexptsneMB.Rd index ca78123..e7eb5c9 100644 --- a/man/plotexptsneMB.Rd +++ b/man/plotexptsneMB.Rd @@ -28,12 +28,3 @@ The t-SNE map representation can also be used to analyze expression of a gene or a group of genes, to investigate cluster specific gene expression patterns } -\examples{ -\donttest{ -data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset -sc <- valuesG1msReduced_treated_MB -sc <- MB_Order(sc, export = FALSE) -g <- 'ENSG00000001460' -plotexptsneMB(sc, g) -} -} diff --git a/man/plotsilhouetteMB.Rd b/man/plotsilhouetteMB.Rd index 477b329..55ec02c 100644 --- a/man/plotsilhouetteMB.Rd +++ b/man/plotsilhouetteMB.Rd @@ -26,8 +26,3 @@ The silhouette provides a representation of how well each point take values between -1 and 1 with higher values reflecting better representation of a point by its cluster. } -\examples{ -data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset -sc <- valuesG1msReduced_treated_MB -plotsilhouetteMB(sc, K=2) -} diff --git a/man/plottSNE.Rd b/man/plottSNE.Rd index bf1c831..6300ef1 100644 --- a/man/plottSNE.Rd +++ b/man/plottSNE.Rd @@ -15,8 +15,3 @@ A plot of t-SNEs. \description{ Visualizing the K-means clusters using tSNE maps } -\examples{ -data(valuesG1msReduced_treated_K) # details: DIscBIO:::prepExampleDataset -sc <- valuesG1msReduced_treated_K -plottSNE(sc) -} diff --git a/man/plottsneMB.Rd b/man/plottsneMB.Rd index 5214204..446a3db 100644 --- a/man/plottsneMB.Rd +++ b/man/plottsneMB.Rd @@ -20,8 +20,3 @@ A plot of t-SNEs. \description{ Visualizing the Model-based clusters using tSNE maps } -\examples{ -data(valuesG1msReduced_treated_MB) # details: DIscBIO:::prepExampleDataset -sc <- valuesG1msReduced_treated_MB -plottsneMB(sc) -} From 5ca11d1e2cdb96d1b903e9a9162d64081529ed00 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Wed, 15 Jul 2020 11:34:32 +0200 Subject: [PATCH 40/99] Reduced dataset --- .Rbuildignore | 2 ++ R/DIscBIO-classes.R | 6 +++--- R/DIscBIO-generic-Clustexp.R | 2 +- R/DIscBIO-generic-Exprmclust.R | 4 ++-- R/DIscBIO-generic-FinalPreprocessing.R | 2 +- R/DIscBIO-generic-FindOutliersKM.R | 2 +- R/DIscBIO-generic-FindOutliersMB.R | 8 ++++---- R/DIscBIO-generic-KMClustDiffGenes.R | 2 +- R/DIscBIO-generic-NoiseFiltering.R | 2 +- R/DIscBIO-generic-Normalizedata.R | 2 +- R/DIscBIO-generic-PCAplotSymbols.R | 2 +- R/DIscBIO-generic-comptSNE.R | 2 +- R/DIscBIO-generic-comptsneMB.R | 6 +++--- R/customConverters.R | 4 ++-- R/datasets.R | 2 +- data/valuesG1msRed.rda | Bin 0 -> 33380 bytes data/valuesG1msTest.rda | Bin 0 -> 19920 bytes tests/testthat/test.DEGfreeze.R | 2 +- tests/testthat/test.integration.R | 2 +- vignettes/DIscBIO-vignette.Rmd | 2 +- 20 files changed, 28 insertions(+), 26 deletions(-) create mode 100644 data/valuesG1msRed.rda create mode 100644 data/valuesG1msTest.rda diff --git a/.Rbuildignore b/.Rbuildignore index c02729d..485cdd7 100644 --- a/.Rbuildignore +++ b/.Rbuildignore @@ -6,6 +6,8 @@ CONTRIBUTING.md DIscBIOlogo.png README.md notebook +data/valuesG1msReduced +data/valuesG1msTest data/valuesG1ms.rda data/genes20k.rda data/pan_indrop_matrix_8000cells_18556genes.rda diff --git a/R/DIscBIO-classes.R b/R/DIscBIO-classes.R index c9a50f0..839f338 100644 --- a/R/DIscBIO-classes.R +++ b/R/DIscBIO-classes.R @@ -52,11 +52,11 @@ #' @importClassesFrom SingleCellExperiment SingleCellExperiment #' #' @examples -#' class(valuesG1msReduced) -#' G1_reclassified <- DISCBIO(valuesG1msReduced) +#' class(valuesG1msRed) +#' G1_reclassified <- DISCBIO(valuesG1msRed) #' class(G1_reclassified) #' str(G1_reclassified, max.level=2) -#' identical(G1_reclassified@expdataAll, valuesG1msReduced) +#' identical(G1_reclassified@expdataAll, valuesG1msRed) #' @export DISCBIO <- setClass( Class = "DISCBIO", diff --git a/R/DIscBIO-generic-Clustexp.R b/R/DIscBIO-generic-Clustexp.R index ac73cfa..b1ac23f 100644 --- a/R/DIscBIO-generic-Clustexp.R +++ b/R/DIscBIO-generic-Clustexp.R @@ -34,7 +34,7 @@ #' @importFrom methods is #' @return The DISCBIO-class object input with the cpart slot filled. #' @examples -#' sc <- DISCBIO(valuesG1msReduced) # changes signature of data +#' sc <- DISCBIO(valuesG1msRed) # changes signature of data #' sc <- Clustexp(sc, cln=3) setGeneric("Clustexp", function(object, clustnr = 20, bootnr = 50, metric = "pearson", do.gap = TRUE, SE.method = "Tibs2001SEmax", diff --git a/R/DIscBIO-generic-Exprmclust.R b/R/DIscBIO-generic-Exprmclust.R index d372805..4a0a0e1 100644 --- a/R/DIscBIO-generic-Exprmclust.R +++ b/R/DIscBIO-generic-Exprmclust.R @@ -19,14 +19,14 @@ #' returns a named list containing the four objects that together correspond #' to the contents of the MBclusters slot. #' @examples -#' sc <- DISCBIO(valuesG1msReduced) +#' sc <- DISCBIO(valuesG1msRed) #' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) #' sc <- Normalizedata( #' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, #' dsn=1, rseed=17000 #' ) #' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- Exprmclust(sc,K = 2) +#' sc <- Exprmclust(sc,K=2) #' print(sc@MBclusters) setGeneric( name = "Exprmclust", diff --git a/R/DIscBIO-generic-FinalPreprocessing.R b/R/DIscBIO-generic-FinalPreprocessing.R index 2c00348..1dcb221 100644 --- a/R/DIscBIO-generic-FinalPreprocessing.R +++ b/R/DIscBIO-generic-FinalPreprocessing.R @@ -8,7 +8,7 @@ #' @param quiet if `TRUE`, intermediary output is suppressed #' @return The DISCBIO-class object input with the FinalGeneList slot filled. #' @examples -#' sc <- DISCBIO(valuesG1msReduced) +#' sc <- DISCBIO(valuesG1msRed) #' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) #' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) setGeneric( diff --git a/R/DIscBIO-generic-FindOutliersKM.R b/R/DIscBIO-generic-FindOutliersKM.R index e51db3c..2ccebb1 100644 --- a/R/DIscBIO-generic-FindOutliersKM.R +++ b/R/DIscBIO-generic-FindOutliersKM.R @@ -23,7 +23,7 @@ #' @return A named vector of the genes containing outlying cells and the number #' of cells on each. #' @examples -#' sc <- DISCBIO(valuesG1msReduced) +#' sc <- DISCBIO(valuesG1msRed) #' sc <- Clustexp(sc, cln=3) # K-means clustering #' Outliers <- FindOutliersKM( #' sc, K=3, outminc=5, outlg=2, probthr=.5*1e-3, thr=2**-(1:40), diff --git a/R/DIscBIO-generic-FindOutliersMB.R b/R/DIscBIO-generic-FindOutliersMB.R index 93ca891..6910e92 100644 --- a/R/DIscBIO-generic-FindOutliersMB.R +++ b/R/DIscBIO-generic-FindOutliersMB.R @@ -23,16 +23,16 @@ #' @return A named vector of the genes containing outlying cells and the number #' of cells on each. #' @examples -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) +#' sc <- DISCBIO(valuesG1msRed) +#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE, plot=FALSE) #' sc <- Normalizedata( #' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, #' dsn=1, rseed=17000 #' ) #' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- Exprmclust(sc) +#' sc <- Exprmclust(sc, K=2) #' FindOutliersMB( -#' sc, K=3, outminc=5, outlg=2, probthr=.5*1e-3, thr=2**-(1:40), +#' sc, K=2, outminc=5, outlg=2, probthr=.5*1e-3, thr=2**-(1:40), #' outdistquant=.75, plot = FALSE, quiet = TRUE #' ) #' diff --git a/R/DIscBIO-generic-KMClustDiffGenes.R b/R/DIscBIO-generic-KMClustDiffGenes.R index c54f2b0..5d043cd 100644 --- a/R/DIscBIO-generic-KMClustDiffGenes.R +++ b/R/DIscBIO-generic-KMClustDiffGenes.R @@ -17,7 +17,7 @@ #' @return A list containing two tables. #' @export #' @examples -#' sc <- DISCBIO(valuesG1msReduced) +#' sc <- DISCBIO(valuesG1msRed) #' sc <- Clustexp(sc, cln=3, quiet=TRUE) # K-means clustering #' KMClustDiffGenes(sc, K=3, fdr=.3, export=FALSE) setGeneric( diff --git a/R/DIscBIO-generic-NoiseFiltering.R b/R/DIscBIO-generic-NoiseFiltering.R index 834f279..2f8a548 100644 --- a/R/DIscBIO-generic-NoiseFiltering.R +++ b/R/DIscBIO-generic-NoiseFiltering.R @@ -27,7 +27,7 @@ #' @note This function should be used only if the dataset has ERCC. #' @return The DISCBIO-class object input with the noiseF slot filled. #' @examples -#' sc <- DISCBIO(valuesG1msReduced) # changes signature of data +#' sc <- DISCBIO(valuesG1msRed) # changes signature of data #' sd_filtered <- NoiseFiltering(sc, export=FALSE) #' str(sd_filtered) setGeneric( diff --git a/R/DIscBIO-generic-Normalizedata.R b/R/DIscBIO-generic-Normalizedata.R index 84b0fc6..8a7db05 100644 --- a/R/DIscBIO-generic-Normalizedata.R +++ b/R/DIscBIO-generic-Normalizedata.R @@ -24,7 +24,7 @@ #' @include DIscBIO-classes.R #' @return The DISCBIO-class object input with the ndata and fdata slots filled. #' @examples -#' sc <- DISCBIO(valuesG1msReduced) # changes signature of data +#' sc <- DISCBIO(valuesG1msRed) # changes signature of data #' #' # In this case this function is used to normalize the reads #' sc_normal <- Normalizedata( diff --git a/R/DIscBIO-generic-PCAplotSymbols.R b/R/DIscBIO-generic-PCAplotSymbols.R index b43a076..391c9d3 100644 --- a/R/DIscBIO-generic-PCAplotSymbols.R +++ b/R/DIscBIO-generic-PCAplotSymbols.R @@ -7,7 +7,7 @@ #' @importFrom graphics legend #' @return Plot of the Principal Components #' @examples -#' sc <- DISCBIO(valuesG1msReduced) +#' sc <- DISCBIO(valuesG1msRed) #' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) #' sc <- Normalizedata( #' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, diff --git a/R/DIscBIO-generic-comptSNE.R b/R/DIscBIO-generic-comptSNE.R index f57e939..045b060 100644 --- a/R/DIscBIO-generic-comptSNE.R +++ b/R/DIscBIO-generic-comptSNE.R @@ -12,7 +12,7 @@ #' @importFrom stats as.dist cor #' @return The DISCBIO-class object input with the tsne slot filled. #' @examples -#' sc <- DISCBIO(valuesG1msReduced) # changes signature of data +#' sc <- DISCBIO(valuesG1msRed) # changes signature of data #' sc <- Clustexp(sc, cln=3) # data must be clustered before plottin #' sc <- comptSNE(sc, max_iter=30) #' head(sc@tsne) diff --git a/R/DIscBIO-generic-comptsneMB.R b/R/DIscBIO-generic-comptsneMB.R index ea7db0e..529b6e0 100644 --- a/R/DIscBIO-generic-comptsneMB.R +++ b/R/DIscBIO-generic-comptsneMB.R @@ -11,14 +11,14 @@ #' @importFrom stats as.dist cor #' @return The DISCBIO-class object input with the MBtsne slot filled. #' @examples -#' sc <- DISCBIO(valuesG1msReduced) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) +#' sc <- DISCBIO(valuesG1msRed) +#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE, plot=FALSE) #' sc <- Normalizedata( #' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, #' dsn=1, rseed=17000 #' ) #' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- Exprmclust(sc) +#' sc <- Exprmclust(sc, K=2) #' sc <- comptsneMB(sc, rseed=15555, max_iter = 1000) #' print(sc@MBtsne) setGeneric( diff --git a/R/customConverters.R b/R/customConverters.R index 37709cb..7364e2c 100644 --- a/R/customConverters.R +++ b/R/customConverters.R @@ -82,7 +82,7 @@ customConvertFeats <- function(x, verbose = TRUE) { #' #' @examples #' g1_sce <- SingleCellExperiment::SingleCellExperiment( -#' list(counts=as.matrix(valuesG1msReduced)) +#' list(counts=as.matrix(valuesG1msRed)) #' ) #' g1_disc <- as.DISCBIO(g1_sce) #' class(g1_disc) @@ -157,7 +157,7 @@ as.DISCBIO <- function(x, ...) { #' @export #' #' @examples -#' g1_disc <- DISCBIO(valuesG1msReduced) +#' g1_disc <- DISCBIO(valuesG1msRed) #' class(g1_disc) #' g1_sce <- DISCBIO2SingleCellExperiment(g1_disc) #' class(g1_sce) diff --git a/R/datasets.R b/R/datasets.R index 87ffd2f..fc26474 100644 --- a/R/datasets.R +++ b/R/datasets.R @@ -6,7 +6,7 @@ #' part of a larger dataset containing 94 single cells. 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b/tests/testthat/test.DIscBIO.IMP.R @@ -4,10 +4,10 @@ context("Data loading and pre-processing") -sc <- DISCBIO(valuesG1msReduced) # Reduced dataset used for testing +sc <- DISCBIO(valuesG1msRed) # Reduced dataset used for testing test_that("Loading datasets generate the expected output", { - expect_equal(dim(valuesG1msReduced), c(1092, 30)) + expect_equal(dim(valuesG1msRed), c(1000, 21)) }) test_that("Data signature changes", { @@ -19,7 +19,7 @@ test_that("Data signature changes", { sc <- NoiseFiltering(sc, plot=FALSE, export=FALSE, quiet=TRUE) test_that("Noise filtering is added", { - expect_equal(length(sc@noiseF), 341) + expect_equal(length(sc@noiseF), 272) }) # In this case this function is used to normalize the reads @@ -30,14 +30,14 @@ sc <- Normalizedata( test_that("Data is normalized", { expect_equal(class(sc@fdata), "data.frame") - expect_output(str(sc@fdata), "1000 obs. of 30 variables") + expect_output(str(sc@fdata), "908 obs. of 21 variables") }) # This function can be used for: 1- filtering and normalizing the dataset that has no ERCC. 2- to normalize and filter genes and cells after the noise filtering. sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE, quiet=TRUE) test_that("Data is normalized", { - expect_equal(dim(sc@fdata), c(341, 30)) + expect_equal(dim(sc@fdata), c(272, 21)) }) # ---------------------------------------------------------------------------- # @@ -51,13 +51,13 @@ sc <- comptSNE(sc, rseed=15555, quiet=TRUE, max_iter=10) test_that("tSNE is computed", { expect_equal(class(sc@tsne), "data.frame") - expect_output(str(sc@tsne), "30 obs. of 2 variables") + expect_output(str(sc@tsne), "21 obs. of 2 variables") }) test_that("Cluster plots output is as expexted", { expect_equivalent( object = Jaccard(sc, Clustering="K-means", K=2, plot=FALSE, R=5), - expected = c(.790, .653), + expected = c(.327, .425), tolerance = .01 ) expect_equal( @@ -74,23 +74,15 @@ Outliers <- FindOutliersKM( sc, K=2, outminc=5, outlg=2, probthr=.5*1e-3, thr=2 ^ (-1:-40), outdistquant=.75, plot = FALSE, quiet = TRUE ) - -# Adjusting outliers -outlg <- round(length(sc@fdata[, 1]) / 200) # The cell will be considered as an outlier if it has a minimum of 0.5% of the number of filtered genes as outlier genes. -Outliers2 <- FindOutliersKM( - sc, K=2, outminc=5, outlg=outlg, probthr=.5*1e-3, thr=2 ^ (-1:-40), - outdistquant=.75, plot = FALSE, quiet = TRUE -) Order <- KmeanOrder(sc, quiet = TRUE, export = FALSE) test_that("Outliers are the expected", { - expect_equivalent(Outliers, c(3, 7, 19)) - expect_equivalent(Outliers2, c(3, 7, 19)) + expect_equivalent(Outliers, c(3, 7, 14)) expect_equivalent( object = Order@kordering, expected = c( - 23, 20, 6, 21, 27, 26, 24, 28, 10, 19, 15, 25, 16, 8, 14, 13, 22, 4, - 17, 2, 3, 18, 11, 29, 9, 5, 12, 1, 30, 7 + 15, 13, 5, 14, 20, 18, 16, 21, 17, 9, 8, 12, 3, 10, 2, 11, 7, 19, + 6, 4, 1 ) ) }) @@ -110,7 +102,7 @@ cdiff2 <- DEGanalysis( # differential expression analysis between two particular clusters. cdiff3 <- DEGanalysis2clust( - sc, Clustering="K-means", K=2, fdr=.15, name="Name", First="CL1", + sc, Clustering="K-means", K=2, fdr=.2, name="Name", First="CL1", Second="CL2", export=FALSE, quiet=TRUE, plot=FALSE, nperms=5, nresamp=2 ) @@ -147,135 +139,12 @@ rpartEVAL <- RpartEVAL( ) test_that("Decision tree elements are defined", { - expect_output(str(DATAforDT), "3 obs. of 30 variables") - expect_s3_class(j48dt, "J48") - expect_identical( - attributes(j48dt)$class, c("J48", "Weka_tree", "Weka_classifier") - ) - expect_identical(j48dt_eval, c(TP = 14, FN = 4, FP = 3, TN = 9)) - expect_s3_class(rpartDT, "rpart") - expect_identical(rpartEVAL, c(TP = 16, FN = 2, FP = 4, TN = 8)) -}) - -# ---------------------------------------------------------------------------- # -# Model-based clustering # -# ---------------------------------------------------------------------------- # - -context("Model-based clustering") - -# Technically, this should be done before Clustexp, but it's ok in practice to -# apply it after K-means because it uses different slots. -sc <- Exprmclust(sc, K=2, quiet=TRUE) - -test_that("Model-based clustering elements are OK", { - expect_identical( - object = names(sc@MBclusters), - expected = c("pcareduceres", "MSTtree", "clusterid", "clucenter") - ) -}) - -sc <- comptsneMB(sc, rseed=15555, quiet=TRUE, max_iter=100) - -test_that("tSNE clustering works fine", { - expect_equal(dim(sc@MBtsne), c(30, 2)) -}) - -# --------------------------------- Outliers --------------------------------- # - -context("MB outliers") - -sc <- Clustexp(sc, cln=2, quiet=TRUE, rseed=17000) - -Outliers <- FindOutliersMB( - sc, K=2, outminc=5, outlg=2, probthr=.5*1e-3, thr=2 ^ (-1:-40), - outdistquant=.75, plot = FALSE, quiet = TRUE -) -outlg <- round(length(sc@fdata[, 1]) / 200) # The cell will be considered as an outlier if it has a minimum of 0.5% of the number of filtered genes as outlier genes. -Outliers2 <- FindOutliersMB( - sc, K=2, outminc=5, outlg=outlg, probthr=.5*1e-3, thr=2 ^ (-1:-40), - outdistquant=.75, plot = FALSE, quiet = TRUE -) - -test_that("MB clustering and outliers work as expected", { - expect_equivalent( - object = Jaccard(sc, Clustering="MB", K=2, plot = FALSE, R=5), - expected = c(.819, .499), - tolerance = 0.01 - ) - expect_equivalent(Outliers, c(3, 4, 7)) - expect_equal(outlg, 2) - expect_equal(Outliers2, c("G1_12" = 3, "G1_18" = 4, "G1_21" = 7)) -}) - -sc <- MB_Order(sc, quiet = TRUE, export = FALSE) -mb_heat <- MBclustheatmap(sc, hmethod="single", plot = FALSE, quiet = TRUE) - -test_that("More MB things are OK", { - expect_equal( - object = sc@MBordering, - expected = c( - 8, 28, 27, 18, 21, 10, 29, 26, 17, 25, 5, 13, 12, 19, 16, 15, 23, - 20, 30, 14, 7, 9, 24, 22, 3, 2, 4, 6, 1, 11 - ) - ) - expect_equal(mb_heat, c(1, 2)) -}) - -# ----------------------------------- DEGs ----------------------------------- # - -context("MB DEGs") - -# Binomial DE analysis -cdiff1 <- MBClustDiffGenes(sc, K=2, fdr=.2, export=FALSE, quiet=TRUE) - -# DE analysis between all clusters -cdiff2 <- DEGanalysis( - sc, Clustering="MB", K=2, fdr=.2, name="Name", export=FALSE, quiet=TRUE, - plot = FALSE, nperms=5, nresamp=2 -) - -# differential expression analysis between particular clusters. -cdiff3 <- DEGanalysis2clust( - sc, Clustering="MB", K=2, fdr=.15, name="Name", First="CL1", Second="CL2", - export = FALSE, plot = FALSE, quiet = TRUE, nperms=5, nresamp=2 -) - -test_that("DEGs are calculated", { - expect_identical( - object = sapply(cdiff1, function(x) class(x)[1]), - expected = c("matrix", "data.frame") - ) - expect_identical( - object = sapply(cdiff2, function(x) class(x)[1]), - expected = c("matrix", "data.frame") - ) - expect_equivalent( - object = sapply(cdiff3, function(x) class(x)[1]), - expected = c("matrix", "data.frame", "data.frame", "data.frame") - ) -}) - -# Decision tree -sigDEG <- cdiff3[[1]] -DATAforDT <- ClassVectoringDT( - sc, Clustering="MB", K=2, First="CL1", Second="CL2", sigDEG, quiet = TRUE -) -j48dt <- J48DT(DATAforDT, quiet = TRUE, plot = FALSE) -j48dt_eval <- J48DTeval( - DATAforDT, num.folds=10, First="CL1", Second="CL2", quiet = TRUE -) -rpartDT <- RpartDT(DATAforDT, quiet = TRUE, plot = FALSE) -rpartEVAL <- RpartEVAL( - DATAforDT, num.folds=10, First="CL1", Second="CL2", quiet = TRUE -) - -test_that("Decision tree elements are defined", { - expect_output(str(DATAforDT), "29 obs. of 30 variables") # used to be 31 + expect_output(str(DATAforDT), "5 obs. of 21 variables") expect_s3_class(j48dt, "J48") expect_identical( attributes(j48dt)$class, c("J48", "Weka_tree", "Weka_classifier") ) - expect_identical(j48dt_eval, c(TP = 21, FN = 2, FP = 4, TN = 3)) + expect_identical(j48dt_eval, c(TP = 7, FN = 4, FP = 5, TN = 5)) expect_s3_class(rpartDT, "rpart") - expect_identical(rpartEVAL, c(TP = 20, FN = 3, FP = 4, TN = 3)) -}) + expect_identical(rpartEVAL, c(TP = 5, FN = 6, FP = 10, TN = 0)) +}) \ No newline at end of file From 31cfd79345a4005fc32f0164b5e766d47a3f89f6 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Wed, 15 Jul 2020 11:35:00 +0200 Subject: [PATCH 42/99] Updated documentation --- man/Clustexp.Rd | 2 +- man/DISCBIO.Rd | 6 +++--- man/DISCBIO2SingleCellExperiment.Rd | 2 +- man/Exprmclust.Rd | 4 ++-- man/FinalPreprocessing.Rd | 2 +- man/FindOutliersKM.Rd | 2 +- man/FindOutliersMB.Rd | 8 ++++---- man/KMClustDiffGenes.Rd | 2 +- man/NoiseFiltering.Rd | 2 +- man/Normalizedata.Rd | 2 +- man/PCAplotSymbols.Rd | 2 +- man/as.DISCBIO.Rd | 2 +- man/comptSNE.Rd | 2 +- man/comptsneMB.Rd | 6 +++--- man/{valuesG1msReduced.Rd => valuesG1msRed.Rd} | 4 ++-- 15 files changed, 24 insertions(+), 24 deletions(-) rename man/{valuesG1msReduced.Rd => valuesG1msRed.Rd} (92%) diff --git a/man/Clustexp.Rd b/man/Clustexp.Rd index 7b5698d..a4350f5 100644 --- a/man/Clustexp.Rd +++ b/man/Clustexp.Rd @@ -77,6 +77,6 @@ This functions performs the initial clustering of the RaceID algorithm. } \examples{ -sc <- DISCBIO(valuesG1msReduced) # changes signature of data +sc <- DISCBIO(valuesG1msRed) # changes signature of data sc <- Clustexp(sc, cln=3) } diff --git a/man/DISCBIO.Rd b/man/DISCBIO.Rd index 421dd10..73ba2fe 100644 --- a/man/DISCBIO.Rd +++ b/man/DISCBIO.Rd @@ -83,9 +83,9 @@ from running the noise filtering or/and the expression filtering.} }} \examples{ -class(valuesG1msReduced) -G1_reclassified <- DISCBIO(valuesG1msReduced) +class(valuesG1msRed) +G1_reclassified <- DISCBIO(valuesG1msRed) class(G1_reclassified) str(G1_reclassified, max.level=2) -identical(G1_reclassified@expdataAll, valuesG1msReduced) +identical(G1_reclassified@expdataAll, valuesG1msRed) } diff --git a/man/DISCBIO2SingleCellExperiment.Rd b/man/DISCBIO2SingleCellExperiment.Rd index fb3f993..0d6ba72 100644 --- a/man/DISCBIO2SingleCellExperiment.Rd +++ b/man/DISCBIO2SingleCellExperiment.Rd @@ -17,7 +17,7 @@ Extract the SingleCellExperiment input data from the corresponding input slot in a DISCBIO-class object } \examples{ -g1_disc <- DISCBIO(valuesG1msReduced) +g1_disc <- DISCBIO(valuesG1msRed) class(g1_disc) g1_sce <- DISCBIO2SingleCellExperiment(g1_disc) class(g1_sce) diff --git a/man/Exprmclust.Rd b/man/Exprmclust.Rd index 6c696b3..87b4f59 100644 --- a/man/Exprmclust.Rd +++ b/man/Exprmclust.Rd @@ -61,13 +61,13 @@ this function first uses principal component analysis (PCA) to clustering on the transformed expression values. } \examples{ -sc <- DISCBIO(valuesG1msReduced) +sc <- DISCBIO(valuesG1msRed) sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) sc <- Normalizedata( sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, dsn=1, rseed=17000 ) sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -sc <- Exprmclust(sc,K = 2) +sc <- Exprmclust(sc,K=2) print(sc@MBclusters) } diff --git a/man/FinalPreprocessing.Rd b/man/FinalPreprocessing.Rd index 2a9f228..e17db70 100644 --- a/man/FinalPreprocessing.Rd +++ b/man/FinalPreprocessing.Rd @@ -37,7 +37,7 @@ The DISCBIO-class object input with the FinalGeneList slot filled. This function generates the final filtered normalized dataset. } \examples{ -sc <- DISCBIO(valuesG1msReduced) +sc <- DISCBIO(valuesG1msRed) sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) } diff --git a/man/FindOutliersKM.Rd b/man/FindOutliersKM.Rd index 7e193b9..f08cd13 100644 --- a/man/FindOutliersKM.Rd +++ b/man/FindOutliersKM.Rd @@ -65,7 +65,7 @@ A named vector of the genes containing outlying cells and the number This functions performs the outlier identification } \examples{ -sc <- DISCBIO(valuesG1msReduced) +sc <- DISCBIO(valuesG1msRed) sc <- Clustexp(sc, cln=3) # K-means clustering Outliers <- FindOutliersKM( sc, K=3, outminc=5, outlg=2, probthr=.5*1e-3, thr=2**-(1:40), diff --git a/man/FindOutliersMB.Rd b/man/FindOutliersMB.Rd index 3236c6d..531abf1 100644 --- a/man/FindOutliersMB.Rd +++ b/man/FindOutliersMB.Rd @@ -65,16 +65,16 @@ A named vector of the genes containing outlying cells and the number This functions performs the outlier identification } \examples{ -sc <- DISCBIO(valuesG1msReduced) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) +sc <- DISCBIO(valuesG1msRed) +sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE, plot=FALSE) sc <- Normalizedata( sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, dsn=1, rseed=17000 ) sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -sc <- Exprmclust(sc) +sc <- Exprmclust(sc, K=2) FindOutliersMB( - sc, K=3, outminc=5, outlg=2, probthr=.5*1e-3, thr=2**-(1:40), + sc, K=2, outminc=5, outlg=2, probthr=.5*1e-3, thr=2**-(1:40), outdistquant=.75, plot = FALSE, quiet = TRUE ) diff --git a/man/KMClustDiffGenes.Rd b/man/KMClustDiffGenes.Rd index 39dea6b..eef6539 100644 --- a/man/KMClustDiffGenes.Rd +++ b/man/KMClustDiffGenes.Rd @@ -60,7 +60,7 @@ A list containing two tables. description } \examples{ -sc <- DISCBIO(valuesG1msReduced) +sc <- DISCBIO(valuesG1msRed) sc <- Clustexp(sc, cln=3, quiet=TRUE) # K-means clustering KMClustDiffGenes(sc, K=3, fdr=.3, export=FALSE) } diff --git a/man/NoiseFiltering.Rd b/man/NoiseFiltering.Rd index 17c34b6..77db8fb 100644 --- a/man/NoiseFiltering.Rd +++ b/man/NoiseFiltering.Rd @@ -75,7 +75,7 @@ Given a matrix or data frame of count data, this function This function should be used only if the dataset has ERCC. } \examples{ -sc <- DISCBIO(valuesG1msReduced) # changes signature of data +sc <- DISCBIO(valuesG1msRed) # changes signature of data sd_filtered <- NoiseFiltering(sc, export=FALSE) str(sd_filtered) } diff --git a/man/Normalizedata.Rd b/man/Normalizedata.Rd index bda1241..a9851f0 100644 --- a/man/Normalizedata.Rd +++ b/man/Normalizedata.Rd @@ -64,7 +64,7 @@ This function allows filtering of genes and cells to be used in the downstream analysis. } \examples{ -sc <- DISCBIO(valuesG1msReduced) # changes signature of data +sc <- DISCBIO(valuesG1msRed) # changes signature of data # In this case this function is used to normalize the reads sc_normal <- Normalizedata( diff --git a/man/PCAplotSymbols.Rd b/man/PCAplotSymbols.Rd index d2d9c2e..9ba0d4a 100644 --- a/man/PCAplotSymbols.Rd +++ b/man/PCAplotSymbols.Rd @@ -22,7 +22,7 @@ Plot of the Principal Components Generates a plot of grouped PCA components } \examples{ -sc <- DISCBIO(valuesG1msReduced) +sc <- DISCBIO(valuesG1msRed) sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) sc <- Normalizedata( sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, diff --git a/man/as.DISCBIO.Rd b/man/as.DISCBIO.Rd index 240704d..85ce67a 100644 --- a/man/as.DISCBIO.Rd +++ b/man/as.DISCBIO.Rd @@ -25,7 +25,7 @@ Additional parameters to pass to `list` include, if x is } \examples{ g1_sce <- SingleCellExperiment::SingleCellExperiment( - list(counts=as.matrix(valuesG1msReduced)) + list(counts=as.matrix(valuesG1msRed)) ) g1_disc <- as.DISCBIO(g1_sce) class(g1_disc) diff --git a/man/comptSNE.Rd b/man/comptSNE.Rd index 73b784c..b832d47 100644 --- a/man/comptSNE.Rd +++ b/man/comptSNE.Rd @@ -45,7 +45,7 @@ This function is used to compute the t-Distributed Stochastic Neighbor Embedding (t-SNE). } \examples{ -sc <- DISCBIO(valuesG1msReduced) # changes signature of data +sc <- DISCBIO(valuesG1msRed) # changes signature of data sc <- Clustexp(sc, cln=3) # data must be clustered before plottin sc <- comptSNE(sc, max_iter=30) head(sc@tsne) diff --git a/man/comptsneMB.Rd b/man/comptsneMB.Rd index 69f88eb..8cd1938 100644 --- a/man/comptsneMB.Rd +++ b/man/comptsneMB.Rd @@ -44,14 +44,14 @@ This function is used to compute the t-Distributed Stochastic Neighbor Embedding (t-SNE). } \examples{ -sc <- DISCBIO(valuesG1msReduced) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) +sc <- DISCBIO(valuesG1msRed) +sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE, plot=FALSE) sc <- Normalizedata( sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, dsn=1, rseed=17000 ) sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -sc <- Exprmclust(sc) +sc <- Exprmclust(sc, K=2) sc <- comptsneMB(sc, rseed=15555, max_iter = 1000) print(sc@MBtsne) } diff --git a/man/valuesG1msReduced.Rd b/man/valuesG1msRed.Rd similarity index 92% rename from man/valuesG1msReduced.Rd rename to man/valuesG1msRed.Rd index 810c8d4..263d4b2 100644 --- a/man/valuesG1msReduced.Rd +++ b/man/valuesG1msRed.Rd @@ -1,8 +1,8 @@ % Generated by roxygen2: do not edit by hand % Please edit documentation in R/datasets.R \docType{data} -\name{valuesG1msReduced} -\alias{valuesG1msReduced} +\name{valuesG1msRed} +\alias{valuesG1msRed} \title{Single-cells data from a myxoid liposarcoma cell line} \description{ A dataset of 30 single cells from a myxoid liposarcoma cell From 5f69b8c9940cef45653050c567b2927d809bc439 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Wed, 15 Jul 2020 11:35:39 +0200 Subject: [PATCH 43/99] Improved handling of NULL internal objects --- R/DIscBIO-generic-DEGanalysis2clust.R | 53 +++++++++++++++------------ R/DIscBIO-generic-Exprmclust.R | 4 ++ 2 files changed, 33 insertions(+), 24 deletions(-) diff --git a/R/DIscBIO-generic-DEGanalysis2clust.R b/R/DIscBIO-generic-DEGanalysis2clust.R index 5af9271..5a9c572 100644 --- a/R/DIscBIO-generic-DEGanalysis2clust.R +++ b/R/DIscBIO-generic-DEGanalysis2clust.R @@ -121,6 +121,7 @@ setMethod( wm <- which.min(delta.table[, 5]) if (delta.table[wm, 5] <= fdr) { w <- which(delta.table[, 5] <= fdr) + if (is.null (w)) stop("No suitable deltas. Try a lower FDR value.") delta <- delta.table[w[1], 1] - 0.001 if (plot) { samr.plot(samr.obj, delta) @@ -193,18 +194,20 @@ setMethod( gene_list <- geneList[, 3] idx_genes <- is.element(gene_list, genes) genes2 <- geneList[idx_genes, ] - FinalDEGsL <- merge( - FinalDEGsL, - genes2, - by.x = "genes", - by.y = "ENSEMBL", - all.x = TRUE - ) - FinalDEGsL[, 3] <- FinalDEGsL[, 11] - FinalDEGsL <- FinalDEGsL[, c(-1, -10, -11)] - FinalDEGsL <- FinalDEGsL[order(FinalDEGsL[, 8]), ] - FinalDEGsL[is.na(FinalDEGsL[, 2]), c(2, 3)] <- - FinalDEGsL[is.na(FinalDEGsL[, 2]), 3] + if (!is.null(FinalDEGsL)) { + FinalDEGsL <- merge( + FinalDEGsL, + genes2, + by.x = "genes", + by.y = "ENSEMBL", + all.x = TRUE + ) + FinalDEGsL[, 3] <- FinalDEGsL[, 11] + FinalDEGsL <- FinalDEGsL[, c(-1, -10, -11)] + FinalDEGsL <- FinalDEGsL[order(FinalDEGsL[, 8]), ] + FinalDEGsL[is.na(FinalDEGsL[, 2]), c(2, 3)] <- + FinalDEGsL[is.na(FinalDEGsL[, 2]), 3] + } if (export) { cat( "The results of DEGs are saved in your directory", @@ -260,18 +263,20 @@ setMethod( gene_list <- geneList[, 3] idx_genes <- is.element(gene_list, genes) genes2 <- geneList[idx_genes, ] - FinalDEGsU <- merge( - FinalDEGsU, - genes2, - by.x = "genes", - by.y = "ENSEMBL", - all.x = TRUE - ) - FinalDEGsU[, 3] <- FinalDEGsU[, 11] - FinalDEGsU <- FinalDEGsU[, c(-1, -10, -11)] - FinalDEGsU <- FinalDEGsU[order(FinalDEGsU[, 8]), ] - FinalDEGsU[is.na(FinalDEGsU[, 2]), c(2, 3)] <- - FinalDEGsU[is.na(FinalDEGsU[, 2]), 3] + if (!is.null(FinalDEGsU)) { + FinalDEGsU <- merge( + FinalDEGsU, + genes2, + by.x = "genes", + by.y = "ENSEMBL", + all.x = TRUE + ) + FinalDEGsU[, 3] <- FinalDEGsU[, 11] + FinalDEGsU <- FinalDEGsU[, c(-1, -10, -11)] + FinalDEGsU <- FinalDEGsU[order(FinalDEGsU[, 8]), ] + FinalDEGsU[is.na(FinalDEGsU[, 2]), c(2, 3)] <- + FinalDEGsU[is.na(FinalDEGsU[, 2]), 3] + } if (export) { cat( "The results of DEGs are saved in your directory", diff --git a/R/DIscBIO-generic-Exprmclust.R b/R/DIscBIO-generic-Exprmclust.R index 4a0a0e1..a7bbe9f 100644 --- a/R/DIscBIO-generic-Exprmclust.R +++ b/R/DIscBIO-generic-Exprmclust.R @@ -77,6 +77,7 @@ definition = function(object, warn = FALSE, verbose = !quiet ) + if (is.null(res)) stop("Unable to cluster. Try a lower value for K.") clusterid <- apply(res$z, 1, which.max) clunum <- res$G } else { @@ -143,6 +144,9 @@ setMethod( warn = FALSE, verbose = !quiet ) + if (is.null(res)) { + stop("Unable to cluster. Try a lower value for K.") + } clusterid <- apply(res$z, 1, which.max) clunum <- res$G } else { From 0ff893f61063f2438b2905964d476c1a71dafed7 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Wed, 15 Jul 2020 11:55:18 +0200 Subject: [PATCH 44/99] Removed images from vignette --- vignettes/DIscBIO-vignette.Rmd | 16 +--------------- 1 file changed, 1 insertion(+), 15 deletions(-) diff --git a/vignettes/DIscBIO-vignette.Rmd b/vignettes/DIscBIO-vignette.Rmd index 596b34b..a4b50a5 100644 --- a/vignettes/DIscBIO-vignette.Rmd +++ b/vignettes/DIscBIO-vignette.Rmd @@ -45,9 +45,6 @@ To account for RNA composition and sequencing depth among samples (single-cells) The key idea in filtering genes is to appoint the genes that manifest abundant variation across samples. Filtering genes is a critical step due to its dramatic impact on the downstream analysis. In case the raw data includes ERCC spike-ins, genes will be filtered based on variability in comparison to a noise level estimated from the ERCC spike-ins according to an algorithm developed by Brennecke et al (Brennecke et al., 2013). This algorithm utilizes the dependence of technical noise on the average read count and fits a model to the ERCC spike-ins. Further gene filtering can be implemented based on gene expression. In case the raw data does not include ERCC spike-ins, genes will be only filtered based on minimum expression in certain number of cells. - -![DataPre-processing](Data-Preprocessing.png){width=80%} - ```{r} sc<- DISCBIO(DataSet) ``` @@ -119,8 +116,6 @@ if (OnlyExpressionFiltering==TRUE){ Cellular clustering is performed according to the gene expression profiles to detect cellular sub-population with unique properties. After clustering, pseudo-temporal ordering is generated to indicate the cellular differentiation degree. -![PSCAN](Clustering.png){width=80%} - ## 2.1. K-means Clustering Rare cell type Identification algorithm (RaceID) was used to cluster the pre-processed data using k-means on a similarity distance matrix, which was based on Pearson correlation and the similarity matrix was computed as “1 – Pearson correlation”. The approach of the proposed clustering, i.e., applying k-means on a similarity distance matrix using the Euclidean metric, improves cluster separation. RaceID estimates the number of clusters by finding the minimal clusters' number at the saturation level of gap statistics, which standardizes the within-cluster dispersion. @@ -140,9 +135,6 @@ The Clustexp() functions has several parameters: - rseed Integer number. Random seed to enforce reproducible clustering results. Default is 17000. - quiet if `TRUE`, intermediate output is suppressed -![DIsccBIO](KM1.png){width=80%} - - ```{r} sc<- Clustexp(sc,cln=2,quiet=TRUE) #### K-means clustering to get three clusters plotGap(sc) ### Plotting gap statisticssc<- Clustexp(sc, clustnr=20,bootnr=50,metric="pearson",do.gap=TRUE,SE.method="Tibs2001SEmax",SE.factor=.25,B.gap=50,cln=K,rseed=17000) @@ -236,8 +228,6 @@ plotExptSNE(sc,g) Differentially expressed genes between individual clusters are identified using the significance analysis of sequencing data (SAMseq), which is a new function in significance analysis of microarrays (Li and Tibshirani 2011) in the samr package v2.0 (Tibshirani et all., 2015). SAMseq is a non-parametric statistical function dependent on Wilcoxon rank statistic that equalizes the sizes of the library by a resampling method accounting for the various sequencing depths. The analysis is implemented over the pure raw dataset that has the unnormalized expression read counts after excluding the ERCCs. Furthermore, DEGs in each cluster comparing to all the remaining clusters are determined using binomial differential expression, which is based on binomial counting statistics. -![DIsccBIO](KM2.png){width=80%} - ## 3.1 Identifying DEGs using SAMseq The user can define DEGs between all clusters generated by either K-means or model based clustering by applying the “DEGanalysis” function. Another alternative is to define DEGs between particular clusters generated by K-means clustering by applying the “DEGanalysis2clust” function. The outcome of these two functions is a list of two components: @@ -304,8 +294,6 @@ In case the user would like to get the names of the significant DEGs, then FS sh There are several methods to identify biomarkers, among them are decision trees and hub detection through networking analysis. The outcome of STRING analysis is stored in tab separated values (TSV) files. These TSV files served as an input to check both the connectivity degree and the betweenness centrality, which reflects the communication flow in the defined PPI networks -![DIsccBIO](KM3.png){width=80%} - Decision trees are one of the most efficient classification techniques in biomarkers discovery. Here we use it to predict the sub-population of a target cell based on transcriptomic data. Two types of decision trees can be performed: classification and regression trees (CART) and J48. The decision tree analysis is implemented over a training dataset, which consisted of the DEGs obtained by either SAMseq or the binomial differential expression. The performance of the generated trees can be evaluated for error estimation by ten-fold cross validation assessment using the "J48DTeval" and "RpartEVAL" functions. The decision tree analysis requires the dataset to be class vectored by applying the “ClassVectoringDT” function. ```{r} @@ -418,9 +406,7 @@ GEA ## 2.2. Model-Based clustering -![DIsccBIO](MB1.png){width=80%} - -Model-based clustering assumes that the data are generated by a model and attempts to recover the original model from the data to define cellular clusters. The steps reproduced here are comparable to the ones on Section [2.1](#21-k-means-clustering), and the output will be suppressed for brevity. +Model-based clustering assumes that the data are generated by a model and attempts to recover the original model from the data to define cellular clusters. The steps reproduced here are comparable to the ones on Section [2.1](#21-k-means-clustering). #### 2.2.1. Defining the Cells in the clusters generated by model-based clustering From cd0a381c666d5051c2799a28620bc995972f6cb3 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Wed, 15 Jul 2020 12:10:07 +0200 Subject: [PATCH 45/99] Improved vignette formatting --- vignettes/DIscBIO-vignette.Rmd | 323 +++++++-------------------------- 1 file changed, 66 insertions(+), 257 deletions(-) diff --git a/vignettes/DIscBIO-vignette.Rmd b/vignettes/DIscBIO-vignette.Rmd index a4b50a5..bdc89ed 100644 --- a/vignettes/DIscBIO-vignette.Rmd +++ b/vignettes/DIscBIO-vignette.Rmd @@ -1,5 +1,5 @@ --- -title: "DIscBIO: a user-friendly pipeline for biomarker discovery in single-cell transcriptomics" +title: "DIscBIO tutorial" author: "Salim Ghannoum" date: "`r Sys.Date()`" output: rmarkdown::html_vignette @@ -8,30 +8,22 @@ vignette: > %\VignetteEngine{knitr::rmarkdown} %\VignetteEncoding{UTF-8} --- - -![DIscBIO](DiscBIO.png){width=80%} - -The pipeline consists of four successive steps: data pre-processing, cellular clustering and pseudo-temporal ordering, determining differential expressed genes and identifying biomarkers. - -## Required Packages - ```{r options, echo=FALSE} knitr::opts_chunk$set(fig.width=7, fig.height=7, eval=FALSE) ``` -```{r} -library(DIscBIO) -library(enrichR) -``` +The pipeline consists of four successive steps: data pre-processing, cellular clustering and pseudo-temporal ordering, determining differential expressed genes and identifying biomarkers. + +This vignette is a reduced version of the interactive notebook available [here](https://mybinder.org/v2/gh/SystemsBiologist/PSCAN/discbio-pub?filepath=DIscBIO.ipynb) ## Loading dataset The dataset should be formatted in a data frame where columns refer to samples and rows refer to genes. - ```{r} -FileName<-data("valuesG1msRed") -DataSet<-get(FileName) +library(DIscBIO) +FileName <- data("valuesG1msRed") +DataSet <- get(FileName) ``` ## 1. Data Pre-processing @@ -46,7 +38,7 @@ The key idea in filtering genes is to appoint the genes that manifest abundant v In case the raw data does not include ERCC spike-ins, genes will be only filtered based on minimum expression in certain number of cells. ```{r} -sc<- DISCBIO(DataSet) +sc <- DISCBIO(DataSet) ``` ### 1.1. Filtering and normalizing the raw data that includes ERCCs @@ -74,19 +66,19 @@ To Finalize the preprocessing the function FinalPreprocessing() should be implem ```{r} -sc<-NoiseFiltering(sc,percentile=0.9, CV=0.2) +sc <- NoiseFiltering(sc,percentile=0.9, CV=0.2) #### Normalizing the reads without any further gene filtering -sc<-Normalizedata(sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, dsn=1, rseed=17000) +sc <- Normalizedata(sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, dsn=1, rseed=17000) #### Additional gene filtering step based on gene expression -#MIinExp<- mean(rowMeans(DataSet,na.rm=TRUE)) +#MIinExp <- mean(rowMeans(DataSet,na.rm=TRUE)) #MIinExp -#MinNumber<- round(length(DataSet[1,])/3) # To be expressed in at least one third of the cells. +#MinNumber <- round(length(DataSet[1,])/3) # To be expressed in at least one third of the cells. #MinNumber -#sc<-Normalizedata(sc, mintotal=1000, minexpr=MIinExp, minnumber=MinNumber, maxexpr=Inf, downsample=FALSE, dsn=1, rseed=17000) +#sc <- Normalizedata(sc, mintotal=1000, minexpr=MIinExp, minnumber=MinNumber, maxexpr=Inf, downsample=FALSE, dsn=1, rseed=17000) -sc<-FinalPreprocessing(sc,GeneFlitering="NoiseF",export=TRUE) ### The GeneFiltering can be either "NoiseF" or"ExpF" +sc <- FinalPreprocessing(sc,GeneFlitering="NoiseF",export=TRUE) ### The GeneFiltering can be either "NoiseF" or"ExpF" ``` ### 1.2. Filtering and normalizing the raw data that does not include ERCCs @@ -95,7 +87,7 @@ To normalize and filter the raw data that does not include ERCCs can be done by To Finalize the preprocessing the function FinalPreprocessing() should be implemented by setting the parameter "GeneFlitering" to ExpF. -### ¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤ Running this cell will overwrite the previous cell ¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤ +**¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤ Running this cell will overwrite the previous cell ¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤** ```{r} @@ -105,10 +97,10 @@ OnlyExpressionFiltering=FALSE if (OnlyExpressionFiltering==TRUE){ MIinExp <- mean(rowMeans(DataSet,na.rm=TRUE)) MIinExp - MinNumber<- round(length(DataSet[1,])/3) # To be expressed in at least one third of the cells. + MinNumber <- round(length(DataSet[1,])/3) # To be expressed in at least one third of the cells. MinNumber - sc<-Normalizedata(sc, mintotal=1000, minexpr=MIinExp, minnumber=MinNumber, maxexpr=Inf, downsample=FALSE, dsn=1, rseed=17000) #### In this case this function is used to filter out genes and cells. - sc<-FinalPreprocessing(sc,GeneFlitering="ExpF",export=TRUE) + sc <- Normalizedata(sc, mintotal=1000, minexpr=MIinExp, minnumber=MinNumber, maxexpr=Inf, downsample=FALSE, dsn=1, rseed=17000) #### In this case this function is used to filter out genes and cells. + sc <- FinalPreprocessing(sc,GeneFlitering="ExpF",export=TRUE) } ``` @@ -136,15 +128,15 @@ The Clustexp() functions has several parameters: - quiet if `TRUE`, intermediate output is suppressed ```{r} -sc<- Clustexp(sc,cln=2,quiet=TRUE) #### K-means clustering to get three clusters -plotGap(sc) ### Plotting gap statisticssc<- Clustexp(sc, clustnr=20,bootnr=50,metric="pearson",do.gap=TRUE,SE.method="Tibs2001SEmax",SE.factor=.25,B.gap=50,cln=K,rseed=17000) +sc <- Clustexp(sc,cln=2,quiet=TRUE) #### K-means clustering to get three clusters +plotGap(sc) ### Plotting gap statisticssc <- Clustexp(sc, clustnr=20,bootnr=50,metric="pearson",do.gap=TRUE,SE.method="Tibs2001SEmax",SE.factor=.25,B.gap=50,cln=K,rseed=17000) ``` #### 2.1.1. Defining the Cells in the clusters generated by k-means clustering ```{r} -sc<- comptSNE(sc,rseed=15555,quiet=TRUE) +sc <- comptSNE(sc,rseed=15555,quiet=TRUE) cat("\t"," Cell-ID"," Cluster Number","\n") sc@cpart ``` @@ -157,11 +149,11 @@ PSCAN enables the robustness assessment of the detected clusters in terms of sta ```{r} # Silhouette of k-means clusters # you might have to run "par(mar=c(6,2,4,2))" first so plot looks better -plotSilhouette(sc,K=2) # K is the number of clusters +plotSilhouette(sc,K=2) # K is the number of clusters ``` ```{r} -Jaccard(sc,Clustering="K-means", K=2, plot=TRUE) # Jaccard of k-means clusters +Jaccard(sc,Clustering="K-means", K=2, plot=TRUE) # Jaccard of k-means clusters ``` #### 2.1.3. Cluster plotting using tSNE maps @@ -172,8 +164,8 @@ Here you visualize the K-means clusters using t-distributed stochastic neighbor ```{r} ############ Plotting K-means clusters plottSNE(sc) -plotKmeansLabelstSNE(sc) # To plot the the ID of the cells in eacj cluster -plotSymbolstSNE(sc,types=sub("(\\_\\d+)$","", names(sc@ndata))) # To plot the the ID of the cells in eacj cluster +plotKmeansLabelstSNE(sc) # To plot the the ID of the cells in eacj cluster +plotSymbolstSNE(sc,types=sub("(\\_\\d+)$","", names(sc@ndata))) # To plot the the ID of the cells in eacj cluster ``` ### Defining outlier cells based on K-means Clustering @@ -184,8 +176,8 @@ In case the user decided to remove outlier cells, the user should set RemovingOu ```{r} -outlg<-round(length(sc@fdata[,1])/200) # The cell will be considered as an outlier if it has a minimum of 0.5% of the number of filtered genes as outlier genes. -Outliers<- FindOutliersKM(sc, K=2, outminc=5,outlg=outlg,probthr=.5*1e-3,thr=2**-(1:40),outdistquant=.75, plot=TRUE, quiet=FALSE) +outlg <- round(length(sc@fdata[,1])/200) # The cell will be considered as an outlier if it has a minimum of 0.5% of the number of filtered genes as outlier genes. +Outliers <- FindOutliersKM(sc, K=2, outminc=5,outlg=outlg,probthr=.5*1e-3,thr=2**-(1:40),outdistquant=.75, plot=TRUE, quiet=FALSE) RemovingOutliers=FALSE # RemovingOutliers=TRUE # Removing the defined outlier cells based on K-means Clustering @@ -203,7 +195,7 @@ if(RemovingOutliers==TRUE){ #### 2.1.4. Cellular pseudo-time ordering based on k-means clusters ```{r} -sc<-KmeanOrder(sc,quiet=FALSE, export=TRUE) +sc <- KmeanOrder(sc,quiet=FALSE, export=TRUE) plotOrderKMtsne(sc) ``` @@ -238,29 +230,29 @@ The user can define DEGs between all clusters generated by either K-means or mod ```{r degKM} ####### differential expression analysis between cluster 1 and cluster 3 of the Model-Based clustering using FDR of 0.05 -cdiff<-DEGanalysis2clust(sc,Clustering="K-means",K=2,fdr=0.05,name="Name",First="CL1",Second="CL2",export=TRUE,quiet=TRUE) +cdiff <- DEGanalysis2clust(sc,Clustering="K-means",K=2,fdr=0.05,name="Name",First="CL1",Second="CL2",export=TRUE,quiet=TRUE) ``` ```{r} #### To show the result table -head(cdiff[[1]]) # The first component -head(cdiff[[2]]) # The second component +head(cdiff[[1]]) # The first component +head(cdiff[[2]]) # The second component ``` ### 3.1.2 Determining DEGs between all clusters -## ¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤ Running this cell will overwrite the previous one ¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤ +**¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤ Running this cell will overwrite the previous one ¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤** ```{r} -cdiff<-DEGanalysis(sc,Clustering="K-means",K=2,fdr=0.05,name="all_clusters",export=TRUE,quiet=TRUE) ####### differential expression analysis between all clusters +cdiff <- DEGanalysis(sc,Clustering="K-means",K=2,fdr=0.05,name="all_clusters",export=TRUE,quiet=TRUE) ####### differential expression analysis between all clusters ``` ```{r} #### To show the result table -head(cdiff[[1]]) # The first component -head(cdiff[[2]]) # The second component +head(cdiff[[1]]) # The first component +head(cdiff[[2]]) # The second component ``` #### 3.2 Identifying DEGs using binomial differential expression @@ -270,10 +262,10 @@ comparison to the ensemble of all cells. It returns a list with a data.frame ele ```{r} -cdiffBinomial<-KMClustDiffGenes(sc,K=2,fdr=.01,,quiet=TRUE) ########## Binomial differential expression analysis +cdiffBinomial <- KMClustDiffGenes(sc,K=2,fdr=.01,,quiet=TRUE) ########## Binomial differential expression analysis #### To show the result table -head(cdiffBinomial[[1]]) # The first component -head(cdiffBinomial[[2]]) # The second component +head(cdiffBinomial[[1]]) # The first component +head(cdiffBinomial[[2]]) # The second component ``` ### Plotting the DEGs @@ -297,18 +289,18 @@ There are several methods to identify biomarkers, among them are decision trees Decision trees are one of the most efficient classification techniques in biomarkers discovery. Here we use it to predict the sub-population of a target cell based on transcriptomic data. Two types of decision trees can be performed: classification and regression trees (CART) and J48. The decision tree analysis is implemented over a training dataset, which consisted of the DEGs obtained by either SAMseq or the binomial differential expression. The performance of the generated trees can be evaluated for error estimation by ten-fold cross validation assessment using the "J48DTeval" and "RpartEVAL" functions. The decision tree analysis requires the dataset to be class vectored by applying the “ClassVectoringDT” function. ```{r} -#sigDEG<-cdiff[[1]] # DEGs gene list from SANseq -sigDEG<-cdiffBinomial[[1]] # DEGs gene list from Binomial analysis +#sigDEG <- cdiff[[1]] # DEGs gene list from SANseq +sigDEG <- cdiffBinomial[[1]] # DEGs gene list from Binomial analysis First="CL1" Second="CL2" -DATAforDT<-ClassVectoringDT(sc,Clustering="K-means",K=3,First=First,Second=Second,sigDEG) +DATAforDT <- ClassVectoringDT(sc,Clustering="K-means",K=3,First=First,Second=Second,sigDEG) ``` ### 4.1. J48 Decision Tree ```{r} -j48dt<-J48DT(DATAforDT) +j48dt <- J48DT(DATAforDT) summary(j48dt) ``` @@ -316,7 +308,7 @@ summary(j48dt) ```{r} -j48dt<-J48DTeval(DATAforDT,num.folds=10,First=First,Second=Second) +j48dt <- J48DTeval(DATAforDT,num.folds=10,First=First,Second=Second) ``` ### 4.2. RPART Decision Tree @@ -330,7 +322,7 @@ rpartDT <- RpartDT(DATAforDT) ```{r} -rpartEVAL<-RpartEVAL(DATAforDT,num.folds=10,First=First,Second=Second) +rpartEVAL <- RpartEVAL(DATAforDT,num.folds=10,First=First,Second=Second) ``` ### 4.3. Networking Analysis @@ -344,18 +336,18 @@ To define protein-protein interactions (PPI) over a list of genes, STRING-api is DEGs="All DEGs" FileName=paste0(DEGs) -data<-cdiffBinomial[[1]] [,2] # DEGs gene list from Binomial analysis (taking only the firat 100 genes) -#data<-cdiff[[1]] [,2] # From the table of the differential expression analysis between all pairs of clusters +data <- cdiffBinomial[[1]] [,2] # DEGs gene list from Binomial analysis (taking only the firat 100 genes) +#data <- cdiff[[1]] [,2] # From the table of the differential expression analysis between all pairs of clusters -ppi<-PPI(data,FileName) +ppi <- PPI(data,FileName) -networking<-NetAnalysis(ppi) +networking <- NetAnalysis(ppi) networking ##### In case the Examine response components=200 and an error "linkmat[i, ]" appeared, that means there are no PPI. ``` ```{r} data=networking[1:25,1] # plotting the network of the top 25 highly connected genes -network<-Networking(data,FileName,plot_width=25, plot_height=10) +network <- Networking(data,FileName,plot_width=25, plot_height=10) ``` ### 4.3.2 Particular set of DEGs @@ -367,15 +359,15 @@ DEGs=cdiffBinomial[[2]][1,4] # Up-regulated genes in cluster 1 (from the Bin #DEGs=cdiff[[2]][1,4] # Up-regulated genes in cluster 1 (from SAMseq) FileName=paste0(DEGs) -data<-read.csv(file=paste0(DEGs),head=TRUE,sep=",") -data<-data[,3] +data <- read.csv(file=paste0(DEGs),head=TRUE,sep=",") +data <- data[,3] -ppi<-PPI(data,FileName) +ppi <- PPI(data,FileName) -networking<-NetAnalysis(ppi) +networking <- NetAnalysis(ppi) networking ##### In case the Examine response components=200 and an error "linkmat[i, ]" appeared, that means there are no PPI. -network<-Networking(data,FileName) +network <- Networking(data,FileName) ``` ### 4.4 Gene Enrichment Analysis @@ -391,16 +383,16 @@ head(dbs) DEGs=cdiffBinomial[[2]][1,4] # Up-regulated genes in cluster 1 (from the Binomial analysis) #DEGs=cdiff[[2]][1,4] # Up-regulated genes in cluster 1 (from SAMseq) -data<-read.csv(file=paste0(DEGs),head=TRUE,sep=",") -data<-as.character(data[,3]) +data <- read.csv(file=paste0(DEGs),head=TRUE,sep=",") +data <- as.character(data[,3]) dbs <- c("KEGG_2013","GO_Molecular_Function_2015", "GO_Biological_Process_2015") enriched <- enrichr(data, dbs) -KEGG_2013<-enriched[[1]][,c(1,2,3,9)] -GO_Molecular_Function_2015<-enriched[[2]][,c(1,2,3,9)] -GO_Biological_Process_2015<-enriched[[3]][,c(1,2,3,9)] +KEGG_2013 <- enriched[[1]][,c(1,2,3,9)] +GO_Molecular_Function_2015 <- enriched[[2]][,c(1,2,3,9)] +GO_Biological_Process_2015 <- enriched[[3]][,c(1,2,3,9)] -GEA<-rbind(KEGG_2013,GO_Molecular_Function_2015,GO_Biological_Process_2015) +GEA <- rbind(KEGG_2013,GO_Molecular_Function_2015,GO_Biological_Process_2015) GEA ``` @@ -429,7 +421,7 @@ PCAplotSymbols(sc) ```{r, eval=FALSE} # Plotting the model-based clusters in tSNE maps -sc<- comptsneMB(sc,rseed=15555,quiet=TRUE) +sc <- comptsneMB(sc,rseed=15555,quiet=TRUE) plottsneMB(sc,K=3) plotMBLabelstSNE(sc) ``` @@ -444,16 +436,16 @@ plotsilhouetteMB(sc,K=3) ``` ```{r, eval=FALSE} -sc<- Clustexp(sc,cln=3,quiet=TRUE) -Jaccard(sc,Clustering="MB", K=3, plot=TRUE) # Jaccard of Model based clusters +sc <- Clustexp(sc,cln=3,quiet=TRUE) +Jaccard(sc,Clustering="MB", K=3, plot=TRUE) # Jaccard of Model based clusters ``` ### Defining outlier cells based on Model-Based Clustering ```{r, eval=FALSE} -outlg<-round(length(sc@fdata[,1])/200) # The cell will be considered as an outlier if it has a minimum of 0.5% of the number of filtered genes as outlier genes. -Outliers<- FindOutliersMB(sc, K=3, outminc=5,outlg=outlg,probthr=.5*1e-3,thr=2**-(1:40),outdistquant=.75, plot=TRUE, quiet=TRUE) +outlg <- round(length(sc@fdata[,1])/200) # The cell will be considered as an outlier if it has a minimum of 0.5% of the number of filtered genes as outlier genes. +Outliers <- FindOutliersMB(sc, K=3, outminc=5,outlg=outlg,probthr=.5*1e-3,thr=2**-(1:40),outdistquant=.75, plot=TRUE, quiet=TRUE) RemovingOutliers=FALSE # RemovingOutliers=TRUE # Removing the defined outlier cells based on K-means Clustering @@ -472,7 +464,7 @@ if(RemovingOutliers==TRUE){ ```{r, eval=FALSE} -sc<-MB_Order(sc,quiet=FALSE, export=TRUE) +sc <- MB_Order(sc,quiet=FALSE, export=TRUE) ``` #### 2.2.4.1 Plotting the pseudo-time ordering in a PCA plot @@ -512,187 +504,4 @@ plotexptsneMB(sc,g) ```{r, eval=FALSE} g <- "ENSG00000010244" #### Plotting the expression of MT-RNR2 PlotMBexpPCA(sc,g) -``` - -# 3. Determining differentially expressed genes (DEGs) - -Differentially expressed genes between individual clusters are identified using the significance analysis of sequencing data (SAMseq), which is a new function in significance analysis of microarrays (Li and Tibshirani 2011) in the samr package v2.0 (Tibshirani et all., 2015). SAMseq is a non-parametric statistical function dependent on Wilcoxon rank statistic that equalizes the sizes of the library by a resampling method accounting for the various sequencing depths. The analysis is implemented over the pure raw dataset that has the unnormalized expression read counts after excluding the ERCCs. Furthermore, DEGs in each cluster comparing to all the remaining clusters are determined using binomial differential expression, which is based on binomial counting statistics. - -![DIsccBIO](MB2.png){width=80%} - -## 3.1 Identifying DEGs using SAMseq - -The user can define DEGs between all clusters generated by either K-means or model based clustering by applying the “DEGanalysis” function. Another alternative is to define DEGs between particular clusters generated by either K-means or model based clustering by applying the “DEGanalysis2clust” function. The outcome of these two functions is a list of two components: -- The first component is a data frame showing the Ensembl gage name and the symbole of the detected DEGs -- The second component is table showing the comparisons, Target cluster, Number of genes and the File name. This component will be used for the downstream analysis. - -### 3.1.1 Determining DEGs between two particular clusters - - -```{r, eval=FALSE} -####### differential expression analysis between cluster 2 and cluster 3 of the Model-Based clustering using FDR of 0.05 -MBcdiff<-DEGanalysis2clust(sc,Clustering="MB",K=3,fdr=0.1,name="Name",First="CL1",Second="CL2",export=TRUE,quiet=FALSE) -#### To show the result table -head(MBcdiff[[1]]) # The first component -head(MBcdiff[[2]]) # The second component -``` - -### 3.1.2 Determining DEGs between all clusters - -## ¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤ Running this cell will overwrite the previous one ¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤ - - -```{r, eval=FALSE} -MBcdiff<-DEGanalysis(sc,Clustering="MB",K=3,fdr=0.05,name="all_clusters",export=TRUE,quiet=TRUE) ####### differential expression analysis between all clusters -#### To show the result table -head(MBcdiff[[1]]) # The first component -head(MBcdiff[[2]]) # The second component -``` - -```{r, eval=FALSE} -#### To show the result table -head(MBcdiff[[1]]) # The first component -head(MBcdiff[[2]]) # The second component -``` - -#### 3.2 Identifying DEGs using binomial differential expression - -The function MBClustDiffGenes identifies differentially regulated genes for each cluster of the Model-Based clustering in -comparison to the ensemble of all cells. It returns a list with a data.frame element for each cluster that contains the mean expression across all cells not in the cluster (mean.ncl) and in the cluster (mean.cl), the fold-change in the cluster versus all remaining cells (fc), and the p-value for differential expression between all cells in a cluster and all remaining cells. The p-value is computed based on the overlap of negative binomials fitted to the count distributions within the two groups akin to DESeq. - - -```{r, eval=FALSE} -MBcdiffBinomial<-MBClustDiffGenes(sc,K=3,fdr=.01,export=TRUE, quiet=TRUE) ########## Binomial differential expression analysis -#### To show the result table -head(MBcdiffBinomial[[1]]) # The first component -head(MBcdiffBinomial[[2]]) # The second component -``` - -### Plotting the DEGs - -Volcano plots are used to readily show the DEGs by plotting significance versus fold-change on the y and x axes, respectively. - - -```{r, eval=FALSE} -name<-MBcdiffBinomial[[2]][2,4] ############ Selecting the "Up-DEG-cluster2.csv " from the DEGs' binomial table ############## -#name<-MBcdiff[[2]][4,6] ############ Selecting the "Up-DEG-cluster2.csv " from the DEGs' SAMseq table ############## -U<-read.csv(file=paste0(name),head=TRUE,sep=",") -Vplot<-VolcanoPlot(U,value=0.001,name=name,FS=.5,fc=0.5) -#### In case the output of this function is an error "Error in plot.window(...): need finite 'ylim' values", the adj should be set to TRUE -### In case the user would like to get the names of the significant DEGs then FS should be set to a value of 0.3 or higher. -``` - -## 4. Identifying biomarkers (decision trees and networking analysis) - -There are several methods to identify biomarkers, among them are decision trees and hub detection through networking analysis. The outcome of STRING analysis is stored in tab separated values (TSV) files. These TSV files served as an input to check both the connectivity degree and the betweenness centrality, which reflects the communication flow in the defined PPI networks - -![DIsccBIO](MB3.png){width=80%} - -Decision trees are one of the most efficient classification techniques in biomarkers discovery. Here we use it to predict the sub-population of a target cell based on transcriptomic data. Two types of decision trees can be performed: classification and regression trees (CART) and J48. The decision tree analysis is implemented over a training dataset, which consisted of the DEGs obtained by either SAMseq or the binomial differential expression. The performance of the generated trees can be evaluated for error estimation by ten-fold cross validation assessment using the "J48DTeval" and "RpartEVAL" functions. -The decision tree analysis requires the dataset to be class vectored by applying the “ClassVectoringDT” function. - - -```{r, eval=FALSE} -sigDEG<-cdiff[[1]] # DEGs gene list from SANseq -#sigDEG<-cdiffBinomial[[1]] # DEGs gene list from Binomial analysis -First="CL1" -Second="CL2" -DATAforDT<-ClassVectoringDT(sc,Clustering="MB",K=3,First=First,Second=Second,sigDEG) -``` - -### 4.1. J48 Decision Tree - - -```{r, eval=FALSE} -j48dt<-J48DT(DATAforDT) -summary(j48dt) -``` - -#### 4.1.1. Evaluating the performance of the J48 Decision Tree - - -```{r, eval=FALSE} -j48dt<-J48DTeval(DATAforDT,num.folds=10,First=First,Second=Second) -``` - -### 4.2. RPART Decision Tree - - -```{r, eval=FALSE} -rpartDT<-RpartDT(DATAforDT) -``` - -#### 4.2.1. Evaluating the performance of the RPART Decision Tree - - -```{r, eval=FALSE} -rpartEVAL<-RpartEVAL(DATAforDT,num.folds=10,First=First,Second=Second) -``` - -### 4.3. Networking Analysis - -To define protein-protein interactions (PPI) over a list of genes, STRING-api is used. The outcome of STRING analysis was stored in tab separated values (TSV) files. These TSV files served as an input to check both the connectivity degree and the betweenness centrality, which reflects the communication flow in the defined PPI networks. - -### 4.3.1 All DEGs - - -```{r, eval=FALSE} -DEGs="All DEGs" -FileName=paste0(DEGs) - -#data<-MBcdiffBinomial[[1]] [,2] # DEGs gene list from Binomial analysis -data<-MBcdiff[[1]] [,2] # From the table of the differential expression analysis between all pairs of clusters - -ppi<-PPI(data,FileName) - -networking<-NetAnalysis(ppi) -networking ##### In case the Examine response components=200 and an error "linkmat[i, ]" appeared, that means there are no PPI. - -network<-Networking(data,FileName) -``` - -### 4.3.2 Particular set of DEGs - - -```{r, eval=FALSE} -############ Selecting the DEGs' table ############## -DEGs=cdiffBinomial[[2]][2,4] # UP-regulated genes in cluster 2 (from the Binomial analysis) -#DEGs=MBcdiff[[2]][2,6] # UP-regulated genes in cluster 2 (from SAMseq) -FileName=paste0(DEGs) - -data<-read.csv(file=paste0(DEGs),head=TRUE,sep=",") -data<-data[,3] - -ppi<-PPI(data,FileName) - -networking<-NetAnalysis(ppi) -networking ##### In case the Examine response components=200 and an error "linkmat[i, ]" appeared, that means there are no PPI. - -network<-Networking(data,FileName) - -``` - -### 4.4 Gene Enrichment Analysis - -```{r, eval=FALSE} -dbs <- listEnrichrDbs() -head(dbs) -#print(dbs) -``` - -```{r, eval=FALSE} -############ Selecting the DEGs' table ############## -DEGs=MBcdiffBinomial[[2]][2,4] # Up-regulated genes in cluster 2 (from the Binomial analysis) -#DEGs=MBcdiff[[2]][2,6] # UP-regulated genes in cluster 2 (from SAMseq) -data<-read.csv(file=paste0(DEGs),head=TRUE,sep=",") -data<-as.character(data[,3]) - -dbs <- c("KEGG_2013","GO_Molecular_Function_2015", "GO_Biological_Process_2015") -enriched <- enrichr(data, dbs) -KEGG_2013<-enriched[[1]][,c(1,2,3,9)] -GO_Molecular_Function_2015<-enriched[[2]][,c(1,2,3,9)] -GO_Biological_Process_2015<-enriched[[3]][,c(1,2,3,9)] - -GEA<-rbind(KEGG_2013,GO_Molecular_Function_2015,GO_Biological_Process_2015) -GEA ``` \ No newline at end of file From 91a8ea03001d6b8873a74a610614ca7f21f51832 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Tue, 21 Jul 2020 10:55:22 +0200 Subject: [PATCH 46/99] Changed cat() to message() --- R/DIscBIO-generic-ClassVectoringDT.R | 9 +- R/DIscBIO-generic-Clustexp.R | 8 +- R/DIscBIO-generic-DEGanalysis.R | 22 +- R/DIscBIO-generic-DEGanalysis2clust.R | 26 +- R/DIscBIO-generic-FinalPreprocessing.R | 61 +- R/DIscBIO-generic-FindOutliersKM.R | 4 +- R/DIscBIO-generic-FindOutliersMB.R | 2 +- R/DIscBIO-generic-NoiseFiltering.R | 28 +- R/J48DTeval.R | 6 +- R/NetAnalysis.R | 16 +- R/Networking.R | 10 +- R/PPI.R | 9 +- R/RpartEVAL.R | 5 +- R/samr-adapted.R | 1288 ++++++++++++------------ man/FinalPreprocessing.Rd | 8 +- vignettes/DIscBIO-vignette.Rmd | 8 +- 16 files changed, 738 insertions(+), 772 deletions(-) diff --git a/R/DIscBIO-generic-ClassVectoringDT.R b/R/DIscBIO-generic-ClassVectoringDT.R index 6aab1a8..9e1f2a1 100644 --- a/R/DIscBIO-generic-ClassVectoringDT.R +++ b/R/DIscBIO-generic-ClassVectoringDT.R @@ -74,13 +74,10 @@ setMethod( gene_names2 <- gene_names[idx_genes] DEGsfilteredDataset <- sg1[gene_names2, ] if (!quiet) { - cat( + message( "The DEGs filtered normalized dataset contains:\n", - "Genes:", - length(DEGsfilteredDataset[, 1]), - "\n", - "cells:", - length(DEGsfilteredDataset[1, ]) + "Genes: ", length(DEGsfilteredDataset[, 1]), "\n", + "cells: ", length(DEGsfilteredDataset[1, ]) ) } G_list = sigDEG diff --git a/R/DIscBIO-generic-Clustexp.R b/R/DIscBIO-generic-Clustexp.R index b1ac23f..e0aa94f 100644 --- a/R/DIscBIO-generic-Clustexp.R +++ b/R/DIscBIO-generic-Clustexp.R @@ -247,11 +247,9 @@ setMethod( ch = calinhara(data, km[[k]]$cluster) ) - if (critout) - cat(k, - " clusters ", - crit[k], - "\n") + if (critout) { + message(k, " clusters ", crit[k], "\n") + } } } if (cluster1) diff --git a/R/DIscBIO-generic-DEGanalysis.R b/R/DIscBIO-generic-DEGanalysis.R index 3199a5e..ae2e637 100644 --- a/R/DIscBIO-generic-DEGanalysis.R +++ b/R/DIscBIO-generic-DEGanalysis.R @@ -91,7 +91,7 @@ setMethod( colnames(dataset) <- Nam if (!quiet) { - cat("The dataset is ready for differential expression analysis") + message("The dataset is ready for differential expression analysis") } num1 <- paste("CL", num, sep = "") clustName <- paste("Cl", num, sep = "") @@ -191,7 +191,7 @@ setMethod( o <- c(1:K) oo <- o[-length(o)] com <- sum(oo) - if (!quiet) cat("Number of comparisons: ", com * 2, "\n") + if (!quiet) message("Number of comparisons: ", com * 2, "\n") comNUM <- paste("comp", c(1:com), sep = "") DEGsTable <- data.frame() DEGsE <- c() @@ -330,11 +330,9 @@ setMethod( FinalDEGsL[is.na(FinalDEGsL[, 2]), c(2, 3)] <- FinalDEGsL[is.na(FinalDEGsL[, 2]), 3] if (!quiet) { - cat( - paste0( - "Low-regulated genes in the ", second[i], - " in ", first[i], " VS ", second[i], "\n" - ) + message( + "Low-regulated genes in the ", second[i], + " in ", first[i], " VS ", second[i], "\n" ) } if (export) { @@ -395,11 +393,9 @@ setMethod( FinalDEGsU[is.na(FinalDEGsU[, 2]), c(2, 3)] <- FinalDEGsU[is.na(FinalDEGsU[, 2]), 3] if (!quiet) { - cat( - paste0( - "Up-regulated genes in the ", second[i], " in ", - first[i], " VS ", second[i], "\n" - ) + message( + "Up-regulated genes in the ", second[i], " in ", + first[i], " VS ", second[i], "\n" ) } if (export) { @@ -440,7 +436,7 @@ setMethod( } if (!quiet & export) { - cat("The results of DEGs are saved in your directory", "\n") + message("The results of DEGs are saved in your directory") } colnames(DEGsTable) <- c( "Comparisons", "Target cluster", "Gene number", "File name", diff --git a/R/DIscBIO-generic-DEGanalysis2clust.R b/R/DIscBIO-generic-DEGanalysis2clust.R index 5a9c572..b3e813d 100644 --- a/R/DIscBIO-generic-DEGanalysis2clust.R +++ b/R/DIscBIO-generic-DEGanalysis2clust.R @@ -209,15 +209,10 @@ setMethod( FinalDEGsL[is.na(FinalDEGsL[, 2]), 3] } if (export) { - cat( - "The results of DEGs are saved in your directory", - "\n" - ) - cat( - paste0( - "Low-regulated genes in the ", Second, " in ", - First, " VS ", Second, "\n" - ) + message("The results of DEGs are saved in your directory") + message( + "Low-regulated genes in the ", Second, " in ", + First, " VS ", Second, "\n" ) write.csv( FinalDEGsL, @@ -278,15 +273,10 @@ setMethod( FinalDEGsU[is.na(FinalDEGsU[, 2]), 3] } if (export) { - cat( - "The results of DEGs are saved in your directory", - "\n" - ) - cat( - paste0( - "Up-regulated genes in the ", Second, " in ", First, - " VS ", Second, "\n" - ) + message("The results of DEGs are saved in your directory") + message( + "Up-regulated genes in the ", Second, " in ", First, + " VS ", Second, "\n" ) write.csv( FinalDEGsU, diff --git a/R/DIscBIO-generic-FinalPreprocessing.R b/R/DIscBIO-generic-FinalPreprocessing.R index 1dcb221..d7495fd 100644 --- a/R/DIscBIO-generic-FinalPreprocessing.R +++ b/R/DIscBIO-generic-FinalPreprocessing.R @@ -6,6 +6,7 @@ #' @param export A logical vector that allows writing the final gene list in #' excel file. Default is TRUE. #' @param quiet if `TRUE`, intermediary output is suppressed +#' @param fileName File name for exporting (if `export = TRUE`) #' @return The DISCBIO-class object input with the FinalGeneList slot filled. #' @examples #' sc <- DISCBIO(valuesG1msRed) @@ -14,7 +15,11 @@ setGeneric( "FinalPreprocessing", function( - object, GeneFlitering = "NoiseF", export = FALSE, quiet = FALSE + object, + GeneFlitering = "NoiseF", + export = FALSE, + quiet = FALSE, + fileName = "filteredDataset" ) standardGeneric("FinalPreprocessing") ) @@ -24,9 +29,7 @@ setGeneric( setMethod( "FinalPreprocessing", signature = "DISCBIO", - definition = function( - object, GeneFlitering, export, quiet = FALSE - ) + definition = function(object, GeneFlitering, export, quiet, fileName) { if (GeneFlitering == "NoiseF") { if (length(object@noiseF) < 1) @@ -40,52 +43,30 @@ setMethod( filteredDataset <- object@fdata[gene_names2, ] object@fdata <- filteredDataset object@FinalGeneList <- rownames(filteredDataset) - - if (!quiet) { - cat( - "The gene filtering method= Noise filtering\n\n", - "The Filtered Normalized dataset contains:\n", - "Genes:", - length(filteredDataset[, 1]), - "\n", - "cells:", - length(filteredDataset[1, ]), - "\n\n" - ) - } - if (export) { - cat( - "The Filtered Normalized dataset was saved as: ", - "filteredDataset.Rdata\n" - ) - save(filteredDataset, file = "filteredDataset.Rdata") - } } if (GeneFlitering == "ExpF") { if (nrow(object@fdata) < 1) stop("run Normalizedata before running FinalPreprocessing") filteredDataset <- object@fdata object@FinalGeneList <- rownames(filteredDataset) - + } + if (!quiet) { + message( + "The gene filtering method = Noise filtering\n\n", + "The Filtered Normalized dataset contains:\n", + "Genes: ", length(filteredDataset[, 1]), "\n", + "cells: ", length(filteredDataset[1, ]), "\n\n" + ) + } + if (export) { + fileNameExt <- paste0(fileName, ".Rdata") if (!quiet) { - cat( - "The gene filtering method= Expression filtering\n\n", - "The Filtered Normalized dataset contains:\n", - "Genes:", - length(filteredDataset[, 1]), - "\n", - "cells:", - length(filteredDataset[1,]), - "\n\n" - ) - } - if (export) { - cat( + message( "The Filtered Normalized dataset was saved as: ", - "filteredDataset.Rdata\n" + fileNameExt ) - save(filteredDataset, file = "filteredDataset.Rdata") } + save(filteredDataset, file = fileNameExt) } return(object) } diff --git a/R/DIscBIO-generic-FindOutliersKM.R b/R/DIscBIO-generic-FindOutliersKM.R index 2ccebb1..7f49b94 100644 --- a/R/DIscBIO-generic-FindOutliersKM.R +++ b/R/DIscBIO-generic-FindOutliersKM.R @@ -273,8 +273,8 @@ setMethod( box() } if (!quiet) { - cat( - "The following cells are considered as outlier cells:", + message( + "The following cells are considered outliers: ", which(object@cpart > K), "\n" ) diff --git a/R/DIscBIO-generic-FindOutliersMB.R b/R/DIscBIO-generic-FindOutliersMB.R index 6910e92..98a58a2 100644 --- a/R/DIscBIO-generic-FindOutliersMB.R +++ b/R/DIscBIO-generic-FindOutliersMB.R @@ -302,7 +302,7 @@ setMethod( box() } if (!quiet) { - cat("The following cells are considered as outlier cells:", + message("The following cells are considered as outlier cells:", which(object@cpart > K), "\n") print(which(object@cpart > K)) diff --git a/R/DIscBIO-generic-NoiseFiltering.R b/R/DIscBIO-generic-NoiseFiltering.R index 2f8a548..84540eb 100644 --- a/R/DIscBIO-generic-NoiseFiltering.R +++ b/R/DIscBIO-generic-NoiseFiltering.R @@ -98,10 +98,10 @@ setMethod( ) if (!quiet) { - cat( + message( "Cut-off value for the ERCCs= ", round(minMeanForFit, digits = 2), - "\n\n" + "\n" ) } @@ -117,7 +117,7 @@ setMethod( ) if (!quiet) { - cat("Coefficients of the fit:", "\n") + message("Coefficients of the fit:") print(fit$coefficients) } @@ -132,9 +132,11 @@ setMethod( total <- var(log(cv2ERCC[useForFit])) if (!quiet) { - cat("Explained variances of log CV^2 values= ", + message( + "Explained variances of log CV^2 values= ", c(round(1 - residual / total, digits = 2)), - "\n\n") + "\n" + ) } ## Pick out genes above noise line @@ -157,17 +159,21 @@ setMethod( genes_test <- sapply(genes_test, paste0, collapse = "") if (!quiet) { - cat("Number of genes that passed the filtering= ", + message( + "Number of genes that passed the filtering = ", length(genes_test), - "\n\n") + "\n" + ) } if (export) { write.csv(genes_test, file = paste0(filename, ".csv")) - cat( - "The filtered gene list was saved as:", - "Noise_filtering_genes_test\n" - ) + if (!quiet) { + message( + "The filtered gene list was saved as ", + paste0(filename, ".csv") + ) + } } if (plot) { diff --git a/R/J48DTeval.R b/R/J48DTeval.R index 670cc5c..8e3758f 100644 --- a/R/J48DTeval.R +++ b/R/J48DTeval.R @@ -26,8 +26,7 @@ J48DTeval <- function( #Start cross validation loop class1 <- levels(class.vec)[1] for (fold in 1:length(segments)) { - if (!quiet) - cat("Fold", fold, "of", length(segments), "\n") + if (!quiet) message("Fold ", fold, " of ", length(segments)) #Define training and test set test.ind <- segments[[fold]] training.set <- exp.df[-test.ind, ] @@ -126,12 +125,11 @@ J48DTeval <- function( j48.mcc <- MCC(j48.confusion.matrix) if (!quiet) { - cat( + message( "J48 SN: ", j48.sn, "\n", "J48 SP: ", j48.sp, "\n", "J48 ACC: ", j48.acc, "\n", "J48 MCC: ", j48.mcc, "\n", - sep = "" ) } return(j48.performance) diff --git a/R/NetAnalysis.R b/R/NetAnalysis.R index 769d27c..896a225 100644 --- a/R/NetAnalysis.R +++ b/R/NetAnalysis.R @@ -35,16 +35,12 @@ NetAnalysis <- function( test.graph.adj <- get.adjacency(gg, sparse = FALSE) test.graph.properties <- GenInd(test.graph.adj) - cat("Number of nodes: ", test.graph.properties$N, "\n") - V(gg) - cat("Number of links: ", test.graph.properties$Ltot, "\n") - E(gg) - cat("Link Density: ", test.graph.properties$LD, "\n") - cat("The connectance of the graph: ", - test.graph.properties$C, - "\n") - cat("Mean Distences", mean_distance(gg), "\n") - cat("Average Path Length", average.path.length(gg), "\n", "\n") + message("Number of nodes: ", test.graph.properties$N) + message("Number of links: ", test.graph.properties$Ltot) + message("Link Density: ", test.graph.properties$LD) + message("The connectance of the graph: ", test.graph.properties$C) + message("Mean Distences", mean_distance(gg)) + message("Average Path Length", average.path.length(gg), "\n") AnalysisTable <- AnalysisTable[order(AnalysisTable[, 2], decreasing = TRUE), ] return(AnalysisTable) diff --git a/R/Networking.R b/R/Networking.R index 5916625..818106e 100644 --- a/R/Networking.R +++ b/R/Networking.R @@ -46,9 +46,11 @@ Networking <- function( species ) ) - cat( - "Examine response components =", status_code(repos), "\t", - "200 means successful", "\n" + message( + "Examine response components =", + status_code(repos), + "\t", + "(200 means successful)", ) y <- repos$request$url if (!is.null(FileName)) { @@ -70,7 +72,7 @@ Networking <- function( plot(0:1, 0:1, type = "n", ann = FALSE, axes = FALSE) rasterImage(Network, 0, 0, 1, 1) - cat( + message( "\n", "You can see the network with high resolution", "by clicking on the following link:", diff --git a/R/PPI.R b/R/PPI.R index cc22cef..b00c188 100644 --- a/R/PPI.R +++ b/R/PPI.R @@ -33,12 +33,9 @@ PPI <- function(data, FileName = NULL, species = "9606") { species ) ) - cat( - "Examine response components =", - status_code(repos), - "\t", - "(200 means successful)", - "\n" + message( + "Examine response components = ", status_code(repos), "\t", + "(200 means successful)", "\n" ) # Process API request content repo_content <- content(repos) diff --git a/R/RpartEVAL.R b/R/RpartEVAL.R index ac11e3a..6f9da1e 100644 --- a/R/RpartEVAL.R +++ b/R/RpartEVAL.R @@ -23,7 +23,7 @@ RpartEVAL <- function(data, num.folds = 10, First = "CL1", Second = "CL2", #Start cross validation loop class1 <- levels(class.vec)[1] for (fold in 1:length(segments)) { - if (!quiet) cat("Fold", fold, "of", length(segments), "\n") + if (!quiet) message("Fold ", fold, " of ", length(segments)) #Define training and test set test.ind <- segments[[fold]] training.set <- exp.df[-test.ind, ] @@ -126,12 +126,11 @@ RpartEVAL <- function(data, num.folds = 10, First = "CL1", Second = "CL2", Rpart.mcc <- MCC(Rpart.confusion.matrix) if (!quiet) { - cat( + message( "Rpart SN: ", Rpart.sn, "\n", "Rpart SP: ", Rpart.sp, "\n", "Rpart ACC: ", Rpart.acc, "\n", "Rpart MCC: ", Rpart.mcc, "\n", - sep = "" ) } return(Rpart.performance) diff --git a/R/samr-adapted.R b/R/samr-adapted.R index 03a0d62..ceb1fc8 100644 --- a/R/samr-adapted.R +++ b/R/samr-adapted.R @@ -117,9 +117,9 @@ sammy <- function (data, resp.type = c("Quantitative", "Two class unpaired", depth = scaling.factors = rep(NA, ncol(x)) scaling.factors = (prod(depth)^(1/length(depth)))/depth if (assay.type == "seq") { - cat("Estimating sequencing depths...", fill = T) + message("Estimating sequencing depths...") depth = samr.estimate.depth(x) - cat("Resampling to get new data matrices...", fill = T) + message("Resampling to get new data matrices...") xresamp = resa(x, depth, nresamp = nresamp) } scaling.factors = (prod(depth)^(1/length(depth)))/depth @@ -148,9 +148,11 @@ sammy <- function (data, resp.type = c("Quantitative", "Two class unpaired", x = junk$x sd.internal = sqrt(rowMeans(junk$sd^2)) if (min(table(y)) == 1) { - cat("", fill = T) - cat("Warning: only one timecourse in one or more classes;\nSAM plot and FDRs will be unreliable; only gene scores are informative", - fill = T) + warning( + "Only one timecourse in one or more classes;\n", + "SAM plot and FDRs will be unreliable;", + "only gene scores are informative" + ) } } if (resp.type == samr.const.twoclass.unpaired.timecourse.response) { @@ -464,7 +466,7 @@ sammy <- function (data, resp.type = c("Quantitative", "Two class unpaired", last = nperms.act } for (b in first:last) { - cat(c("perm=", b), fill = TRUE) + message(c("perm = ", b)) if (assay.type == "array") { xstar <- x } @@ -626,18 +628,18 @@ sammy <- function (data, resp.type = c("Quantitative", "Two class unpaired", #' @param x data matrix. nrow=#gene, ncol=#sample #' @return depth: estimated sequencing depth. a vector with len sample. samr.estimate.depth <- function(x) { - iter <- 5 - cmeans <- colSums(x)/sum(x) - for (i in 1:iter) { - n0 <- rowSums(x) %*% t(cmeans) - prop <- rowSums((x - n0)^2/(n0 + 1e-08)) - qs <- quantile(prop, c(0.25, 0.75)) - keep <- (prop >= qs[1]) & (prop <= qs[2]) - cmeans <- colMeans(x[keep, ]) - cmeans <- cmeans/sum(cmeans) - } - depth <- cmeans/mean(cmeans) - return(depth) + iter <- 5 + cmeans <- colSums(x)/sum(x) + for (i in 1:iter) { + n0 <- rowSums(x) %*% t(cmeans) + prop <- rowSums((x - n0)^2/(n0 + 1e-08)) + qs <- quantile(prop, c(0.25, 0.75)) + keep <- (prop >= qs[1]) & (prop <= qs[2]) + cmeans <- colMeans(x[keep, ]) + cmeans <- cmeans/sum(cmeans) + } + depth <- cmeans/mean(cmeans) + return(depth) } #' @title Resampling @@ -648,20 +650,20 @@ samr.estimate.depth <- function(x) { #' @description Corresponds to `samr::resample` #' @importFrom stats rpois runif resa <- function(x, d, nresamp = 20) { - ng <- nrow(x) - ns <- ncol(x) - dbar <- exp(mean(log(d))) - xresamp <- array(0, dim = c(ng, ns, nresamp)) - for (k in 1:nresamp) { - for (j in 1:ns) { - xresamp[, j, k] <- rpois(n = ng, lambda = (dbar/d[j]) * - x[, j]) + runif(ng) * 0.1 - } - } - for (k in 1:nresamp) { - xresamp[, , k] <- t(rankcols(t(xresamp[, , k]))) - } - return(xresamp) + ng <- nrow(x) + ns <- ncol(x) + dbar <- exp(mean(log(d))) + xresamp <- array(0, dim = c(ng, ns, nresamp)) + for (k in 1:nresamp) { + for (j in 1:ns) { + xresamp[, j, k] <- rpois(n = ng, lambda = (dbar/d[j]) * + x[, j]) + runif(ng) * 0.1 + } + } + for (k in 1:nresamp) { + xresamp[, , k] <- t(rankcols(t(xresamp[, , k]))) + } + return(xresamp) } #' @title Rank columns @@ -672,15 +674,15 @@ resa <- function(x, d, nresamp = 20) { #' DEGanalysis functions to freeze in large datasets. #' @param x x rankcols <- function(x) { - # ranks the elements within each col of the matrix x - # and returns these ranks in a matrix - n = nrow(x) - p = ncol(x) - mode(n) = "integer" - mode(p) = "integer" - mode(x) = "double" - xr <- apply(x, 2, rank) - return(xr) + # ranks the elements within each col of the matrix x + # and returns these ranks in a matrix + n = nrow(x) + p = ncol(x) + mode(n) = "integer" + mode(p) = "integer" + mode(x) = "double" + xr <- apply(x, 2, rank) + return(xr) } #' @title Check format @@ -688,75 +690,75 @@ rankcols <- function(x) { #' @param resp.type resp type #' @param censoring.status censoring status check.format <- function(y, resp.type, censoring.status = NULL) { - # here i do some format checks for the input data$y - # note that checks for time course data are done in the - # parse function for time course; - # we then check the output from the parser in this function - if (resp.type == samr.const.twoclass.unpaired.response | - resp.type == samr.const.twoclass.unpaired.timecourse.response) { - if (sum(y == 1) + sum(y == 2) != length(y)) { - stop(paste("Error in input response data: response type ", - resp.type, " specified; values must be 1 or 2")) - } - } - if (resp.type == samr.const.twoclass.paired.response | resp.type == - samr.const.twoclass.paired.timecourse.response) { - if (sum(y) != 0) { - stop(paste("Error in input response data: response type ", - resp.type, " specified; values must be -1, 1, -2, 2, etc")) - } - if (sum(table(y[y > 0]) != abs(table(y[y < 0])))) { - stop(paste("Error in input response data: response type ", - resp.type, " specified; values must be -1, 1, -2, 2, etc")) - } - } - if (resp.type == samr.const.oneclass.response | resp.type == - samr.const.oneclass.timecourse.response) { - if (sum(y == 1) != length(y)) { - stop(paste("Error in input response data: response type ", - resp.type, " specified; values must all be 1")) - } - } - if (resp.type == samr.const.multiclass.response) { - tt = table(y) - nc = length(tt) - if (sum(y <= nc & y > 0) < length(y)) { - stop(paste("Error in input response data: response type ", - resp.type, " specified; values must be 1,2, ... number of classes")) - } - for (k in 1:nc) { - if (sum(y == k) < 2) { - stop(paste("Error in input response data: response type ", - resp.type, " specified; there must be >1 sample per class")) - } - } - } - if (resp.type == samr.const.quantitative.response) { - if (!is.numeric(y)) { - stop(paste("Error in input response data: response type", - resp.type, " specified; values must be numeric")) - } - } - if (resp.type == samr.const.survival.response) { - if (is.null(censoring.status)) { - stop(paste("Error in input response data: response type ", - resp.type, " specified; error in censoring indicator")) - } - if (!is.numeric(y) | sum(y < 0) > 0) { - stop(paste("Error in input response data: response type ", - resp.type, " specified; survival times must be numeric and nonnegative")) - if (sum(censoring.status == 0) + sum(censoring.status == - 1) != length(censoring.status)) { - stop(paste("Error in input response data: response type ", - resp.type, " specified; censoring indicators must be 0 (censored) or 1 (failed)")) - } - } - if (sum(censoring.status == 1) < 1) { - stop(paste("Error in input response data: response type ", - resp.type, " specified; there are no uncensored observations")) - } - } - return() + # here i do some format checks for the input data$y + # note that checks for time course data are done in the + # parse function for time course; + # we then check the output from the parser in this function + if (resp.type == samr.const.twoclass.unpaired.response | + resp.type == samr.const.twoclass.unpaired.timecourse.response) { + if (sum(y == 1) + sum(y == 2) != length(y)) { + stop(paste("Error in input response data: response type ", + resp.type, " specified; values must be 1 or 2")) + } + } + if (resp.type == samr.const.twoclass.paired.response | resp.type == + samr.const.twoclass.paired.timecourse.response) { + if (sum(y) != 0) { + stop(paste("Error in input response data: response type ", + resp.type, " specified; values must be -1, 1, -2, 2, etc")) + } + if (sum(table(y[y > 0]) != abs(table(y[y < 0])))) { + stop(paste("Error in input response data: response type ", + resp.type, " specified; values must be -1, 1, -2, 2, etc")) + } + } + if (resp.type == samr.const.oneclass.response | resp.type == + samr.const.oneclass.timecourse.response) { + if (sum(y == 1) != length(y)) { + stop(paste("Error in input response data: response type ", + resp.type, " specified; values must all be 1")) + } + } + if (resp.type == samr.const.multiclass.response) { + tt = table(y) + nc = length(tt) + if (sum(y <= nc & y > 0) < length(y)) { + stop(paste("Error in input response data: response type ", + resp.type, " specified; values must be 1,2, ... number of classes")) + } + for (k in 1:nc) { + if (sum(y == k) < 2) { + stop(paste("Error in input response data: response type ", + resp.type, " specified; there must be >1 sample per class")) + } + } + } + if (resp.type == samr.const.quantitative.response) { + if (!is.numeric(y)) { + stop(paste("Error in input response data: response type", + resp.type, " specified; values must be numeric")) + } + } + if (resp.type == samr.const.survival.response) { + if (is.null(censoring.status)) { + stop(paste("Error in input response data: response type ", + resp.type, " specified; error in censoring indicator")) + } + if (!is.numeric(y) | sum(y < 0) > 0) { + stop(paste("Error in input response data: response type ", + resp.type, " specified; survival times must be numeric and nonnegative")) + if (sum(censoring.status == 0) + sum(censoring.status == + 1) != length(censoring.status)) { + stop(paste("Error in input response data: response type ", + resp.type, " specified; censoring indicators must be 0 (censored) or 1 (failed)")) + } + } + if (sum(censoring.status == 1) < 1) { + stop(paste("Error in input response data: response type ", + resp.type, " specified; there are no uncensored observations")) + } + } + return() } #' @title Twoclass Wilcoxon statistics @@ -764,54 +766,54 @@ check.format <- function(y, resp.type, censoring.status = NULL) { #' @param y outcome vector of values 1 and 2 #' @return the statistic. wilcoxon.unpaired.seq.func <- function(xresamp, y) { - tt <- rep(0, dim(xresamp)[1]) - for (i in 1:dim(xresamp)[3]) { - tt <- tt + rowSums(xresamp[, y == 2, i]) - sum(y == 2) * - (length(y) + 1)/2 - } - tt <- tt/dim(xresamp)[3] - return(list(tt = tt, numer = tt, sd = rep(1, length(tt)))) + tt <- rep(0, dim(xresamp)[1]) + for (i in 1:dim(xresamp)[3]) { + tt <- tt + rowSums(xresamp[, y == 2, i]) - sum(y == 2) * + (length(y) + 1)/2 + } + tt <- tt/dim(xresamp)[3] + return(list(tt = tt, numer = tt, sd = rep(1, length(tt)))) } wilcoxon.paired.seq.func <- function(xresamp, y) { - tt <- rep(0, dim(xresamp)[1]) - for (i in 1:dim(xresamp)[3]) { - tt <- tt + rowSums(xresamp[, y > 0, i]) - sum(y > 0) * - (length(y) + 1)/2 - } - tt <- tt/dim(xresamp)[3] - return(list(tt = tt, numer = tt, sd = rep(1, length(tt)))) + tt <- rep(0, dim(xresamp)[1]) + for (i in 1:dim(xresamp)[3]) { + tt <- tt + rowSums(xresamp[, y > 0, i]) - sum(y > 0) * + (length(y) + 1)/2 + } + tt <- tt/dim(xresamp)[3] + return(list(tt = tt, numer = tt, sd = rep(1, length(tt)))) } getperms = function(y, nperms) { - total.perms = factorial(length(y)) - if (total.perms <= nperms) { - perms = permute(1:length(y)) - all.perms.flag = 1 - nperms.act = total.perms - } - if (total.perms > nperms) { - perms = matrix(NA, nrow = nperms, ncol = length(y)) - for (i in 1:nperms) { - perms[i, ] = sample(1:length(y), size = length(y)) - } - all.perms.flag = 0 - nperms.act = nperms - } - return(list(perms = perms, all.perms.flag = all.perms.flag, - nperms.act = nperms.act)) + total.perms = factorial(length(y)) + if (total.perms <= nperms) { + perms = permute(1:length(y)) + all.perms.flag = 1 + nperms.act = total.perms + } + if (total.perms > nperms) { + perms = matrix(NA, nrow = nperms, ncol = length(y)) + for (i in 1:nperms) { + perms[i, ] = sample(1:length(y), size = length(y)) + } + all.perms.flag = 0 + nperms.act = nperms + } + return(list(perms = perms, all.perms.flag = all.perms.flag, + nperms.act = nperms.act)) } foldchange.twoclass = function(x, y, logged2) { - # if(logged2){x=2^x} - m1 <- rowMeans(x[, y == 1, drop = F]) - m2 <- rowMeans(x[, y == 2, drop = F]) - if (!logged2) { - fc = m2/m1 - } - if (logged2) { - fc = 2^{ - m2 - m1 - } - } - return(fc) + # if(logged2){x=2^x} + m1 <- rowMeans(x[, y == 1, drop = F]) + m2 <- rowMeans(x[, y == 2, drop = F]) + if (!logged2) { + fc = m2/m1 + } + if (logged2) { + fc = 2^{ + m2 - m1 + } + } + return(fc) } #' @title Foldchange of twoclass unpaired sequencing data #' @importFrom matrixStats rowMedians @@ -820,559 +822,559 @@ foldchange.twoclass = function(x, y, logged2) { #' @param depth depth foldchange.seq.twoclass.unpaired <- function(x, y, depth) { - x.norm <- scale(x, center = F, scale = depth) + 1e-08 - fc <- rowMedians(x.norm[, y == 2])/rowMedians(x.norm[, y == - 1]) - return(fc) + x.norm <- scale(x, center = F, scale = depth) + 1e-08 + fc <- rowMedians(x.norm[, y == 2])/rowMedians(x.norm[, y == + 1]) + return(fc) } foldchange.seq.twoclass.paired <- function(x, y, depth) { - nc <- ncol(x)/2 - o1 <- o2 <- rep(0, nc) - for (j in 1:nc) { - o1[j] <- which(y == -j) - o2[j] <- which(y == j) - } - x.norm <- scale(x, center = F, scale = depth) + 1e-08 - d <- x.norm[, o2, drop = F]/x.norm[, o1, drop = F] - fc <- rowMedians(d, na.rm = T) - return(fc) + nc <- ncol(x)/2 + o1 <- o2 <- rep(0, nc) + for (j in 1:nc) { + o1[j] <- which(y == -j) + o2[j] <- which(y == j) + } + x.norm <- scale(x, center = F, scale = depth) + 1e-08 + d <- x.norm[, o2, drop = F]/x.norm[, o1, drop = F] + fc <- rowMedians(d, na.rm = T) + return(fc) } permute <- function(elem) { - # generates all perms of the vector elem - if (!missing(elem)) { - if (length(elem) == 2) - return(matrix(c(elem, elem[2], elem[1]), nrow = 2)) - last.matrix <- permute(elem[-1]) - dim.last <- dim(last.matrix) - new.matrix <- matrix(0, nrow = dim.last[1] * (dim.last[2] + - 1), ncol = dim.last[2] + 1) - for (row in 1:(dim.last[1])) { - for (col in 1:(dim.last[2] + 1)) new.matrix[row + - (col - 1) * dim.last[1], ] <- insert.value(last.matrix[row, - ], elem[1], col) - } - return(new.matrix) - } - else cat("Usage: permute(elem)\n\twhere elem is a vector\n") + # generates all perms of the vector elem + if (!missing(elem)) { + if (length(elem) == 2) + return(matrix(c(elem, elem[2], elem[1]), nrow = 2)) + last.matrix <- permute(elem[-1]) + dim.last <- dim(last.matrix) + new.matrix <- matrix(0, nrow = dim.last[1] * (dim.last[2] + + 1), ncol = dim.last[2] + 1) + for (row in 1:(dim.last[1])) { + for (col in 1:(dim.last[2] + 1)) new.matrix[row + + (col - 1) * dim.last[1], ] <- insert.value(last.matrix[row, + ], elem[1], col) + } + return(new.matrix) + } + else message("Usage: permute(elem)\n\twhere elem is a vector") } insert.value <- function(vec, newval, pos) { - if (pos == 1) - return(c(newval, vec)) - lvec <- length(vec) - if (pos > lvec) - return(c(vec, newval)) - return(c(vec[1:pos - 1], newval, vec[pos:lvec])) + if (pos == 1) + return(c(newval, vec)) + lvec <- length(vec) + if (pos > lvec) + return(c(vec, newval)) + return(c(vec[1:pos - 1], newval, vec[pos:lvec])) } parse.block.labels.for.2classes = function(y) { - #this only works for 2 class case- having form jBlockn, - # where j=1 or 2 - n = length(y) - y.act = rep(NA, n) - blocky = rep(NA, n) - for (i in 1:n) { - blocky[i] = as.numeric(substring(y[i], 7, nchar(y[i]))) - y.act[i] = as.numeric(substring(y[i], 1, 1)) - } - return(list(y.act = y.act, blocky = blocky)) + #this only works for 2 class case- having form jBlockn, + # where j=1 or 2 + n = length(y) + y.act = rep(NA, n) + blocky = rep(NA, n) + for (i in 1:n) { + blocky[i] = as.numeric(substring(y[i], 7, nchar(y[i]))) + y.act[i] = as.numeric(substring(y[i], 1, 1)) + } + return(list(y.act = y.act, blocky = blocky)) } parse.time.labels.and.summarize.data = function(x, - y, resp.type, time.summary.type) { - # parse time labels, and summarize time data for each - # person, via a slope or area - # does some error checking too - n = length(y) - last5char = rep(NA, n) - last3char = rep(NA, n) - for (i in 1:n) { - last3char[i] = substring(y[i], nchar(y[i]) - 2, nchar(y[i])) - last5char[i] = substring(y[i], nchar(y[i]) - 4, nchar(y[i])) - } - if (sum(last3char == "End") != sum(last5char == "Start")) { - stop("Error in format of time course data: a Start or End tag is missing") - } - y.act = rep(NA, n) - timey = rep(NA, n) - person.id = rep(NA, n) - k = 1 - end.flag = FALSE - person.id[1] = 1 - if (substring(y[1], nchar(y[1]) - 4, nchar(y[1])) != "Start") { - stop("Error in format of time course data: first cell should have a Start tag") - } - for (i in 1:n) { - cat(i) - j = 1 - while (substring(y[i], j, j) != "T") { - j = j + 1 - } - end.of.y = j - 1 - y.act[i] = as.numeric(substring(y[i], 1, end.of.y)) - timey[i] = substring(y[i], end.of.y + 5, nchar(y[i])) - if (nchar(timey[i]) > 3 & substring(timey[i], nchar(timey[i]) - - 2, nchar(timey[i])) == "End") { - end.flag = TRUE - timey[i] = substring(timey[i], 1, nchar(timey[i]) - - 3) - } - if (nchar(timey[i]) > 3 & substring(timey[i], nchar(timey[i]) - - 4, nchar(timey[i])) == "Start") { - timey[i] = substring(timey[i], 1, nchar(timey[i]) - - 5) - } - if (i < n & !end.flag) { - person.id[i + 1] = k - } - if (i < n & end.flag) { - k = k + 1 - person.id[i + 1] = k - } - end.flag = FALSE - } - timey = as.numeric(timey) - # do a check that the format was correct - tt = table(person.id, y.act) - junk = function(x) { - sum(x != 0) - } - if (sum(apply(tt, 1, junk) != 1) > 0) { - num = (1:nrow(tt))[apply(tt, 1, junk) > 1] - stop(paste("Error in format of time course data, timecourse #", - as.character(num))) - } - npeople = length(unique(person.id)) - newx = matrix(NA, nrow = nrow(x), ncol = npeople) - sd = matrix(NA, nrow = nrow(x), ncol = npeople) - for (j in 1:npeople) { - jj = person.id == j - tim = timey[jj] - xc = t(scale(t(x[, jj, drop = F]), center = TRUE, scale = FALSE)) - if (time.summary.type == "slope") { - junk = quantitative.func(xc, tim - mean(tim)) - newx[, j] = junk$numer - sd[, j] = junk$sd - } - if (time.summary.type == "signed.area") { - junk = timearea.func(x[, jj, drop = F], tim) - newx[, j] = junk$numer - sd[, j] = junk$sd - } - } - y.unique = y.act[!duplicated(person.id)] - return(list(y = y.unique, x = newx, sd = sd)) + y, resp.type, time.summary.type) { + # parse time labels, and summarize time data for each + # person, via a slope or area + # does some error checking too + n = length(y) + last5char = rep(NA, n) + last3char = rep(NA, n) + for (i in 1:n) { + last3char[i] = substring(y[i], nchar(y[i]) - 2, nchar(y[i])) + last5char[i] = substring(y[i], nchar(y[i]) - 4, nchar(y[i])) + } + if (sum(last3char == "End") != sum(last5char == "Start")) { + stop("Error in format of time course data: a Start or End tag is missing") + } + y.act = rep(NA, n) + timey = rep(NA, n) + person.id = rep(NA, n) + k = 1 + end.flag = FALSE + person.id[1] = 1 + if (substring(y[1], nchar(y[1]) - 4, nchar(y[1])) != "Start") { + stop("Error in format of time course data: first cell should have a Start tag") + } + for (i in 1:n) { + message(i) + j = 1 + while (substring(y[i], j, j) != "T") { + j = j + 1 + } + end.of.y = j - 1 + y.act[i] = as.numeric(substring(y[i], 1, end.of.y)) + timey[i] = substring(y[i], end.of.y + 5, nchar(y[i])) + if (nchar(timey[i]) > 3 & substring(timey[i], nchar(timey[i]) - + 2, nchar(timey[i])) == "End") { + end.flag = TRUE + timey[i] = substring(timey[i], 1, nchar(timey[i]) - + 3) + } + if (nchar(timey[i]) > 3 & substring(timey[i], nchar(timey[i]) - + 4, nchar(timey[i])) == "Start") { + timey[i] = substring(timey[i], 1, nchar(timey[i]) - + 5) + } + if (i < n & !end.flag) { + person.id[i + 1] = k + } + if (i < n & end.flag) { + k = k + 1 + person.id[i + 1] = k + } + end.flag = FALSE + } + timey = as.numeric(timey) + # do a check that the format was correct + tt = table(person.id, y.act) + junk = function(x) { + sum(x != 0) + } + if (sum(apply(tt, 1, junk) != 1) > 0) { + num = (1:nrow(tt))[apply(tt, 1, junk) > 1] + stop(paste("Error in format of time course data, timecourse #", + as.character(num))) + } + npeople = length(unique(person.id)) + newx = matrix(NA, nrow = nrow(x), ncol = npeople) + sd = matrix(NA, nrow = nrow(x), ncol = npeople) + for (j in 1:npeople) { + jj = person.id == j + tim = timey[jj] + xc = t(scale(t(x[, jj, drop = F]), center = TRUE, scale = FALSE)) + if (time.summary.type == "slope") { + junk = quantitative.func(xc, tim - mean(tim)) + newx[, j] = junk$numer + sd[, j] = junk$sd + } + if (time.summary.type == "signed.area") { + junk = timearea.func(x[, jj, drop = F], tim) + newx[, j] = junk$numer + sd[, j] = junk$sd + } + } + y.unique = y.act[!duplicated(person.id)] + return(list(y = y.unique, x = newx, sd = sd)) } ttest.func <- function(x, y, s0 = 0, sd = NULL) { - n1 <- sum(y == 1) - n2 <- sum(y == 2) - p <- nrow(x) - m1 <- rowMeans(x[, y == 1, drop = F]) - m2 <- rowMeans(x[, y == 2, drop = F]) - if (is.null(sd)) { - sd <- sqrt(((n2 - 1) * varr(x[, y == 2], meanx = m2) + - (n1 - 1) * varr(x[, y == 1], meanx = m1)) * (1/n1 + - 1/n2)/(n1 + n2 - 2)) - } - numer <- m2 - m1 - dif.obs <- (numer)/(sd + s0) - return(list(tt = dif.obs, numer = numer, sd = sd)) + n1 <- sum(y == 1) + n2 <- sum(y == 2) + p <- nrow(x) + m1 <- rowMeans(x[, y == 1, drop = F]) + m2 <- rowMeans(x[, y == 2, drop = F]) + if (is.null(sd)) { + sd <- sqrt(((n2 - 1) * varr(x[, y == 2], meanx = m2) + + (n1 - 1) * varr(x[, y == 1], meanx = m1)) * (1/n1 + + 1/n2)/(n1 + n2 - 2)) + } + numer <- m2 - m1 + dif.obs <- (numer)/(sd + s0) + return(list(tt = dif.obs, numer = numer, sd = sd)) } wilcoxon.func <- function(x, y, s0 = 0) { - n1 <- sum(y == 1) - n2 <- sum(y == 2) - p = nrow(x) - r2 = rowSums(t(apply(x, 1, rank))[, y == 2, drop = F]) - numer = r2 - (n2/2) * (n2 + 1) - (n1 * n2)/2 - sd = sqrt(n1 * n2 * (n1 + n2 + 1)/12) - tt = (numer)/(sd + s0) - return(list(tt = tt, numer = numer, sd = rep(sd, p))) + n1 <- sum(y == 1) + n2 <- sum(y == 2) + p = nrow(x) + r2 = rowSums(t(apply(x, 1, rank))[, y == 2, drop = F]) + numer = r2 - (n2/2) * (n2 + 1) - (n1 * n2)/2 + sd = sqrt(n1 * n2 * (n1 + n2 + 1)/12) + tt = (numer)/(sd + s0) + return(list(tt = tt, numer = numer, sd = rep(sd, p))) } onesample.ttest.func <- function(x, y, s0 = 0, sd = NULL) { - n <- length(y) - x <- x * matrix(y, nrow = nrow(x), ncol = ncol(x), byrow = TRUE) - m <- rowMeans(x) - if (is.null(sd)) { - sd <- sqrt(varr(x, meanx = m)/n) - } - dif.obs <- m/(sd + s0) - return(list(tt = dif.obs, numer = m, sd = sd)) + n <- length(y) + x <- x * matrix(y, nrow = nrow(x), ncol = ncol(x), byrow = TRUE) + m <- rowMeans(x) + if (is.null(sd)) { + sd <- sqrt(varr(x, meanx = m)/n) + } + dif.obs <- m/(sd + s0) + return(list(tt = dif.obs, numer = m, sd = sd)) } patterndiscovery.func = function(x, s0 = 0, eigengene.number = 1) { - a = mysvd(x, n.components = eigengene.number) - v = a$v[, eigengene.number] - # here we try to guess the most interpretable orientation - # for the eigengene - om = abs(a$u[, eigengene.number]) > quantile(abs(a$u[, eigengene.number]), - 0.95) - if (median(a$u[om, eigengene.number]) < 0) { - v = -1 * v - } - aa = quantitative.func(x, v, s0 = s0) - eigengene = cbind(1:nrow(a$v), v) - dimnames(eigengene) = list(NULL, c("sample number", "value")) - return(list(tt = aa$tt, numer = aa$numer, sd = aa$sd, eigengene = eigengene)) + a = mysvd(x, n.components = eigengene.number) + v = a$v[, eigengene.number] + # here we try to guess the most interpretable orientation + # for the eigengene + om = abs(a$u[, eigengene.number]) > quantile(abs(a$u[, eigengene.number]), + 0.95) + if (median(a$u[om, eigengene.number]) < 0) { + v = -1 * v + } + aa = quantitative.func(x, v, s0 = s0) + eigengene = cbind(1:nrow(a$v), v) + dimnames(eigengene) = list(NULL, c("sample number", "value")) + return(list(tt = aa$tt, numer = aa$numer, sd = aa$sd, eigengene = eigengene)) } paired.ttest.func <- function(x, y, s0 = 0, sd = NULL) { - nc <- ncol(x)/2 - o <- 1:nc - o1 <- rep(0, ncol(x)/2) - o2 <- o1 - for (j in 1:nc) { - o1[j] <- (1:ncol(x))[y == -o[j]] - } - for (j in 1:nc) { - o2[j] <- (1:ncol(x))[y == o[j]] - } - d <- x[, o2, drop = F] - x[, o1, drop = F] - su <- x[, o2, drop = F] + x[, o1, drop = F] - if (is.matrix(d)) { - m <- rowMeans(d) - } - if (!is.matrix(d)) { - m <- mean(d) - } - if (is.null(sd)) { - if (is.matrix(d)) { - sd <- sqrt(varr(d, meanx = m)/nc) - } - if (!is.matrix(d)) { - sd <- sqrt(var(d)/nc) - } - } - dif.obs <- m/(sd + s0) - return(list(tt = dif.obs, numer = m, sd = sd)) + nc <- ncol(x)/2 + o <- 1:nc + o1 <- rep(0, ncol(x)/2) + o2 <- o1 + for (j in 1:nc) { + o1[j] <- (1:ncol(x))[y == -o[j]] + } + for (j in 1:nc) { + o2[j] <- (1:ncol(x))[y == o[j]] + } + d <- x[, o2, drop = F] - x[, o1, drop = F] + su <- x[, o2, drop = F] + x[, o1, drop = F] + if (is.matrix(d)) { + m <- rowMeans(d) + } + if (!is.matrix(d)) { + m <- mean(d) + } + if (is.null(sd)) { + if (is.matrix(d)) { + sd <- sqrt(varr(d, meanx = m)/nc) + } + if (!is.matrix(d)) { + sd <- sqrt(var(d)/nc) + } + } + dif.obs <- m/(sd + s0) + return(list(tt = dif.obs, numer = m, sd = sd)) } cox.func <- function(x, y, censoring.status, s0 = 0) { - # find the index matrix - Dn <- sum(censoring.status == 1) - Dset <- c(1:ncol(x))[censoring.status == 1] # the set of observed - ind <- matrix(0, ncol(x), Dn) - # get the matrix - for (i in 1:Dn) { - ind[y > y[Dset[i]] - 1e-08, i] <- 1/sum(y > y[Dset[i]] - - 1e-08) - } - ind.sums <- rowSums(ind) - x.ind <- x %*% ind - # get the derivatives - numer <- x %*% (censoring.status - ind.sums) - sd <- sqrt((x * x) %*% ind.sums - rowSums(x.ind * x.ind)) - tt <- numer/(sd + s0) - return(list(tt = tt, numer = numer, sd = sd)) + # find the index matrix + Dn <- sum(censoring.status == 1) + Dset <- c(1:ncol(x))[censoring.status == 1] # the set of observed + ind <- matrix(0, ncol(x), Dn) + # get the matrix + for (i in 1:Dn) { + ind[y > y[Dset[i]] - 1e-08, i] <- 1/sum(y > y[Dset[i]] - + 1e-08) + } + ind.sums <- rowSums(ind) + x.ind <- x %*% ind + # get the derivatives + numer <- x %*% (censoring.status - ind.sums) + sd <- sqrt((x * x) %*% ind.sums - rowSums(x.ind * x.ind)) + tt <- numer/(sd + s0) + return(list(tt = tt, numer = numer, sd = sd)) } multiclass.func <- function(x, y, s0 = 0) { - ##assumes y is coded 1,2... - nn <- table(y) - m <- matrix(0, nrow = nrow(x), ncol = length(nn)) - v <- m - for (j in 1:length(nn)) { - m[, j] <- rowMeans(x[, y == j]) - v[, j] <- (nn[j] - 1) * varr(x[, y == j], meanx = m[, - j]) - } - mbar <- rowMeans(x) - mm <- m - matrix(mbar, nrow = length(mbar), ncol = length(nn)) - fac <- (sum(nn)/prod(nn)) - scor <- sqrt(fac * (apply(matrix(nn, nrow = nrow(m), ncol = ncol(m), - byrow = TRUE) * mm * mm, 1, sum))) - sd <- sqrt(rowSums(v) * (1/sum(nn - 1)) * sum(1/nn)) - tt <- scor/(sd + s0) - mm.stand = t(scale(t(mm), center = FALSE, scale = sd)) - return(list(tt = tt, numer = scor, sd = sd, stand.contrasts = mm.stand)) + ##assumes y is coded 1,2... + nn <- table(y) + m <- matrix(0, nrow = nrow(x), ncol = length(nn)) + v <- m + for (j in 1:length(nn)) { + m[, j] <- rowMeans(x[, y == j]) + v[, j] <- (nn[j] - 1) * varr(x[, y == j], meanx = m[, + j]) + } + mbar <- rowMeans(x) + mm <- m - matrix(mbar, nrow = length(mbar), ncol = length(nn)) + fac <- (sum(nn)/prod(nn)) + scor <- sqrt(fac * (apply(matrix(nn, nrow = nrow(m), ncol = ncol(m), + byrow = TRUE) * mm * mm, 1, sum))) + sd <- sqrt(rowSums(v) * (1/sum(nn - 1)) * sum(1/nn)) + tt <- scor/(sd + s0) + mm.stand = t(scale(t(mm), center = FALSE, scale = sd)) + return(list(tt = tt, numer = scor, sd = sd, stand.contrasts = mm.stand)) } est.s0 <- function(tt, sd, s0.perc = seq(0, 1, by = 0.05)) { - ## estimate s0 (exchangeability) factor for denominator. - ## returns the actual estimate s0 (not a percentile) - br = unique(quantile(sd, seq(0, 1, len = 101))) - nbr = length(br) - a <- cut(sd, br, labels = F) - a[is.na(a)] <- 1 - cv.sd <- rep(0, length(s0.perc)) - for (j in 1:length(s0.perc)) { - w <- quantile(sd, s0.perc[j]) - w[j == 1] <- 0 - tt2 <- tt * sd/(sd + w) - tt2[tt2 == Inf] = NA - sds <- rep(0, nbr - 1) - for (i in 1:(nbr - 1)) { - sds[i] <- stats::mad(tt2[a == i], na.rm = TRUE) - } - cv.sd[j] <- sqrt(var(sds))/mean(sds) - } - o = (1:length(s0.perc))[cv.sd == min(cv.sd)] - # we don;t allow taking s0.hat to be 0th percentile when - # min sd is 0 - s0.hat = quantile(sd[sd != 0], s0.perc[o]) - return(list(s0.perc = s0.perc, cv.sd = cv.sd, s0.hat = s0.hat)) + ## estimate s0 (exchangeability) factor for denominator. + ## returns the actual estimate s0 (not a percentile) + br = unique(quantile(sd, seq(0, 1, len = 101))) + nbr = length(br) + a <- cut(sd, br, labels = F) + a[is.na(a)] <- 1 + cv.sd <- rep(0, length(s0.perc)) + for (j in 1:length(s0.perc)) { + w <- quantile(sd, s0.perc[j]) + w[j == 1] <- 0 + tt2 <- tt * sd/(sd + w) + tt2[tt2 == Inf] = NA + sds <- rep(0, nbr - 1) + for (i in 1:(nbr - 1)) { + sds[i] <- stats::mad(tt2[a == i], na.rm = TRUE) + } + cv.sd[j] <- sqrt(var(sds))/mean(sds) + } + o = (1:length(s0.perc))[cv.sd == min(cv.sd)] + # we don;t allow taking s0.hat to be 0th percentile when + # min sd is 0 + s0.hat = quantile(sd[sd != 0], s0.perc[o]) + return(list(s0.perc = s0.perc, cv.sd = cv.sd, s0.hat = s0.hat)) } permute.rows <- function(x) { - dd <- dim(x) - n <- dd[1] - p <- dd[2] - mm <- runif(length(x)) + rep(seq(n) * 10, rep(p, n)) - matrix(t(x)[order(mm)], n, p, byrow = TRUE) + dd <- dim(x) + n <- dd[1] + p <- dd[2] + mm <- runif(length(x)) + rep(seq(n) * 10, rep(p, n)) + matrix(t(x)[order(mm)], n, p, byrow = TRUE) } foldchange.paired = function(x, y, logged2) { - # if(logged2){x=2^x} - nc <- ncol(x)/2 - o <- 1:nc - o1 <- rep(0, ncol(x)/2) - o2 <- o1 - for (j in 1:nc) { - o1[j] <- (1:ncol(x))[y == -o[j]] - } - for (j in 1:nc) { - o2[j] <- (1:ncol(x))[y == o[j]] - } - if (!logged2) { - d <- x[, o2, drop = F]/x[, o1, drop = F] - } - if (logged2) { - d <- x[, o2, drop = F] - x[, o1, drop = F] - } - if (!logged2) { - fc <- rowMeans(d) - } - if (logged2) { - fc <- 2^rowMeans(d) - } - return(fc) + # if(logged2){x=2^x} + nc <- ncol(x)/2 + o <- 1:nc + o1 <- rep(0, ncol(x)/2) + o2 <- o1 + for (j in 1:nc) { + o1[j] <- (1:ncol(x))[y == -o[j]] + } + for (j in 1:nc) { + o2[j] <- (1:ncol(x))[y == o[j]] + } + if (!logged2) { + d <- x[, o2, drop = F]/x[, o1, drop = F] + } + if (logged2) { + d <- x[, o2, drop = F] - x[, o1, drop = F] + } + if (!logged2) { + fc <- rowMeans(d) + } + if (logged2) { + fc <- 2^rowMeans(d) + } + return(fc) } foldchange.seq.twoclass.unpaired <- function(x, y, depth) { - x.norm <- scale(x, center = F, scale = depth) + 1e-08 - fc <- rowMedians(x.norm[, y == 2])/rowMedians(x.norm[, y == - 1]) - return(fc) + x.norm <- scale(x, center = F, scale = depth) + 1e-08 + fc <- rowMedians(x.norm[, y == 2])/rowMedians(x.norm[, y == + 1]) + return(fc) } integer.base.b <- function(x, b = 2) { - xi <- as.integer(x) - if (xi == 0) { - return(0) - } - if (any(is.na(xi) | ((x - xi) != 0))) - print(list(ERROR = "x not integer", x = x)) - N <- length(x) - xMax <- max(x) - ndigits <- (floor(logb(xMax, base = 2)) + 1) - Base.b <- array(NA, dim = c(N, ndigits)) - for (i in 1:ndigits) { - #i <- 1 - Base.b[, ndigits - i + 1] <- (x%%b) - x <- (x%/%b) - } - if (N == 1) - Base.b[1, ] - else Base.b + xi <- as.integer(x) + if (xi == 0) { + return(0) + } + if (any(is.na(xi) | ((x - xi) != 0))) + print(list(ERROR = "x not integer", x = x)) + N <- length(x) + xMax <- max(x) + ndigits <- (floor(logb(xMax, base = 2)) + 1) + Base.b <- array(NA, dim = c(N, ndigits)) + for (i in 1:ndigits) { + #i <- 1 + Base.b[, ndigits - i + 1] <- (x%%b) + x <- (x%/%b) + } + if (N == 1) + Base.b[1, ] + else Base.b } varr <- function(x, meanx = NULL) { - n <- ncol(x) - p <- nrow(x) - Y <- matrix(1, nrow = n, ncol = 1) - if (is.null(meanx)) { - meanx <- rowMeans(x) - } - ans <- rep(1, p) - xdif <- x - meanx %*% t(Y) - ans <- (xdif^2) %*% rep(1/(n - 1), n) - ans <- drop(ans) - return(ans) + n <- ncol(x) + p <- nrow(x) + Y <- matrix(1, nrow = n, ncol = 1) + if (is.null(meanx)) { + meanx <- rowMeans(x) + } + ans <- rep(1, p) + xdif <- x - meanx %*% t(Y) + ans <- (xdif^2) %*% rep(1/(n - 1), n) + ans <- drop(ans) + return(ans) } quantitative.func <- function(x, y, s0 = 0) { - # regression of x on y - my = mean(y) - yy <- y - my - temp <- x %*% yy - mx = rowMeans(x) - syy = sum(yy^2) - scor <- temp/syy - b0hat <- mx - scor * my - ym = matrix(y, nrow = nrow(x), ncol = ncol(x), byrow = T) - xhat <- matrix(b0hat, nrow = nrow(x), ncol = ncol(x)) + ym * - matrix(scor, nrow = nrow(x), ncol = ncol(x)) - sigma <- sqrt(rowSums((x - xhat)^2)/(ncol(xhat) - 2)) - sd <- sigma/sqrt(syy) - tt <- scor/(sd + s0) - return(list(tt = tt, numer = scor, sd = sd)) + # regression of x on y + my = mean(y) + yy <- y - my + temp <- x %*% yy + mx = rowMeans(x) + syy = sum(yy^2) + scor <- temp/syy + b0hat <- mx - scor * my + ym = matrix(y, nrow = nrow(x), ncol = ncol(x), byrow = T) + xhat <- matrix(b0hat, nrow = nrow(x), ncol = ncol(x)) + ym * + matrix(scor, nrow = nrow(x), ncol = ncol(x)) + sigma <- sqrt(rowSums((x - xhat)^2)/(ncol(xhat) - 2)) + sd <- sigma/sqrt(syy) + tt <- scor/(sd + s0) + return(list(tt = tt, numer = scor, sd = sd)) } timearea.func <- function(x, y, s0 = 0) { - n <- ncol(x) - xx <- 0.5 * (x[, 2:n] + x[, 1:(n - 1)]) * matrix(diff(y), - nrow = nrow(x), ncol = n - 1, byrow = T) - numer <- rowMeans(xx) - sd <- sqrt(varr(xx, meanx = numer)/n) - tt <- numer/sqrt(sd + s0) - return(list(tt = tt, numer = numer, sd = sd)) + n <- ncol(x) + xx <- 0.5 * (x[, 2:n] + x[, 1:(n - 1)]) * matrix(diff(y), + nrow = nrow(x), ncol = n - 1, byrow = T) + numer <- rowMeans(xx) + sd <- sqrt(varr(xx, meanx = numer)/n) + tt <- numer/sqrt(sd + s0) + return(list(tt = tt, numer = numer, sd = sd)) } cox.seq.func <- function(xresamp, y, censoring.status) { - # get the dimensions - ng <- dim(xresamp)[1] - ns <- dim(xresamp)[2] - # prepare for the calculation - # find the index matrix - Dn <- sum(censoring.status == 1) - Dset <- c(1:ns)[censoring.status == 1] # the set of died - ind <- matrix(0, ns, Dn) - # get the matrix - for (i in 1:Dn) { - ind[y >= y[Dset[i]] - 1e-08, i] <- 1/sum(y >= y[Dset[i]] - - 1e-08) - } - ind.sums <- rowSums(ind) - # calculate the score statistic - tt <- apply(xresamp, 3, function(x, cen.ind, ind.para, ind.sums.para) { - dev1 <- x %*% cen.ind - x.ind <- x %*% ind.para - dev2 <- (x * x) %*% ind.sums.para - rowSums(x.ind * x.ind) - dev1/(sqrt(dev2) + 1e-08) - }, (censoring.status - ind.sums), ind, ind.sums) - tt <- rowMeans(tt) - return(list(tt = tt, numer = tt, sd = rep(1, length(tt)))) + # get the dimensions + ng <- dim(xresamp)[1] + ns <- dim(xresamp)[2] + # prepare for the calculation + # find the index matrix + Dn <- sum(censoring.status == 1) + Dset <- c(1:ns)[censoring.status == 1] # the set of died + ind <- matrix(0, ns, Dn) + # get the matrix + for (i in 1:Dn) { + ind[y >= y[Dset[i]] - 1e-08, i] <- 1/sum(y >= y[Dset[i]] - + 1e-08) + } + ind.sums <- rowSums(ind) + # calculate the score statistic + tt <- apply(xresamp, 3, function(x, cen.ind, ind.para, ind.sums.para) { + dev1 <- x %*% cen.ind + x.ind <- x %*% ind.para + dev2 <- (x * x) %*% ind.sums.para - rowSums(x.ind * x.ind) + dev1/(sqrt(dev2) + 1e-08) + }, (censoring.status - ind.sums), ind, ind.sums) + tt <- rowMeans(tt) + return(list(tt = tt, numer = tt, sd = rep(1, length(tt)))) } compute.block.perms = function(y, blocky, nperms) { - # y are the data (eg class label 1 vs 2; or -1,1, -2,2 for - # paired data) - # blocky are the block labels (abs(y) for paired daatr) - ny = length(y) - nblocks = length(unique(blocky)) - tab = table(blocky) - total.nperms = prod(factorial(tab)) - # block.perms is a list of all possible permutations - block.perms = vector("list", nblocks) - # first enumerate all perms, when possible - if (total.nperms <= nperms) { - all.perms.flag = 1 - nperms.act = total.nperms - for (i in 1:nblocks) { - block.perms[[i]] = permute(y[blocky == i]) - } - kk = 0:(factorial(max(tab))^nblocks - 1) - #the rows of the matrix outerm runs through the 'outer - # product' - # first we assume that all blocks have max(tab) members; - # then we remove rows of outerm that - # are illegal (ie when a block has fewer members) - outerm = matrix(0, nrow = length(kk), ncol = nblocks) - for (i in 1:length(kk)) { - kkkk = integer.base.b(kk[i], b = factorial(max(tab))) - if (length(kkkk) > nblocks) { - kkkk = kkkk[(length(kkkk) - nblocks + 1):length(kkkk)] - } - outerm[i, (nblocks - length(kkkk) + 1):nblocks] = kkkk - } - outerm = outerm + 1 - # now remove rows that are illegal perms - ind = rep(TRUE, nrow(outerm)) - for (j in 1:ncol(outerm)) { - ind = ind & outerm[, j] <= factorial(tab[j]) - } - outerm = outerm[ind, , drop = F] - # finally, construct permutation matrix from outer product - permsy = matrix(NA, nrow = total.nperms, ncol = ny) - for (i in 1:total.nperms) { - junk = NULL - for (j in 1:nblocks) { - junk = c(junk, block.perms[[j]][outerm[i, j], - ]) - } - permsy[i, ] = junk - } - } - # next handle case when there are too many perms to enumerate - if (total.nperms > nperms) { - all.perms.flag = 0 - nperms.act = nperms - permsy = NULL - block.perms = vector("list", nblocks) - for (j in 1:nblocks) { - block.perms[[j]] = sample.perms(y[blocky == j], nperms = nperms) - } - for (j in 1:nblocks) { - permsy = cbind(permsy, block.perms[[j]]) - } - } - return(list(permsy = permsy, all.perms.flag = all.perms.flag, - nperms.act = nperms.act)) + # y are the data (eg class label 1 vs 2; or -1,1, -2,2 for + # paired data) + # blocky are the block labels (abs(y) for paired daatr) + ny = length(y) + nblocks = length(unique(blocky)) + tab = table(blocky) + total.nperms = prod(factorial(tab)) + # block.perms is a list of all possible permutations + block.perms = vector("list", nblocks) + # first enumerate all perms, when possible + if (total.nperms <= nperms) { + all.perms.flag = 1 + nperms.act = total.nperms + for (i in 1:nblocks) { + block.perms[[i]] = permute(y[blocky == i]) + } + kk = 0:(factorial(max(tab))^nblocks - 1) + #the rows of the matrix outerm runs through the 'outer + # product' + # first we assume that all blocks have max(tab) members; + # then we remove rows of outerm that + # are illegal (ie when a block has fewer members) + outerm = matrix(0, nrow = length(kk), ncol = nblocks) + for (i in 1:length(kk)) { + kkkk = integer.base.b(kk[i], b = factorial(max(tab))) + if (length(kkkk) > nblocks) { + kkkk = kkkk[(length(kkkk) - nblocks + 1):length(kkkk)] + } + outerm[i, (nblocks - length(kkkk) + 1):nblocks] = kkkk + } + outerm = outerm + 1 + # now remove rows that are illegal perms + ind = rep(TRUE, nrow(outerm)) + for (j in 1:ncol(outerm)) { + ind = ind & outerm[, j] <= factorial(tab[j]) + } + outerm = outerm[ind, , drop = F] + # finally, construct permutation matrix from outer product + permsy = matrix(NA, nrow = total.nperms, ncol = ny) + for (i in 1:total.nperms) { + junk = NULL + for (j in 1:nblocks) { + junk = c(junk, block.perms[[j]][outerm[i, j], + ]) + } + permsy[i, ] = junk + } + } + # next handle case when there are too many perms to enumerate + if (total.nperms > nperms) { + all.perms.flag = 0 + nperms.act = nperms + permsy = NULL + block.perms = vector("list", nblocks) + for (j in 1:nblocks) { + block.perms[[j]] = sample.perms(y[blocky == j], nperms = nperms) + } + for (j in 1:nblocks) { + permsy = cbind(permsy, block.perms[[j]]) + } + } + return(list(permsy = permsy, all.perms.flag = all.perms.flag, + nperms.act = nperms.act)) } sample.perms <- function(elem, nperms) { - # randomly generates nperms of the vector elem - res = permute.rows(matrix(elem, nrow = nperms, ncol = length(elem), - byrow = T)) - return(res) + # randomly generates nperms of the vector elem + res = permute.rows(matrix(elem, nrow = nperms, ncol = length(elem), + byrow = T)) + return(res) } mysvd <- function(x, n.components = NULL) { - # finds PCs of matrix x - p <- nrow(x) - n <- ncol(x) - # center the observations (rows) - feature.means <- rowMeans(x) - x <- t(scale(t(x), center = feature.means, scale = F)) - if (is.null(n.components)) { - n.components = min(n, p) - } - if (p > n) { - a <- eigen(t(x) %*% x) - v <- a$vec[, 1:n.components, drop = FALSE] - d <- sqrt(a$val[1:n.components, drop = FALSE]) - u <- scale(x %*% v, center = FALSE, scale = d) - return(list(u = u, d = d, v = v)) - } - else { - junk <- svd(x, LINPACK = TRUE) - nc = min(ncol(junk$u), n.components) - return(list(u = junk$u[, 1:nc], d = junk$d[1:nc], v = junk$v[, - 1:nc])) - } + # finds PCs of matrix x + p <- nrow(x) + n <- ncol(x) + # center the observations (rows) + feature.means <- rowMeans(x) + x <- t(scale(t(x), center = feature.means, scale = F)) + if (is.null(n.components)) { + n.components = min(n, p) + } + if (p > n) { + a <- eigen(t(x) %*% x) + v <- a$vec[, 1:n.components, drop = FALSE] + d <- sqrt(a$val[1:n.components, drop = FALSE]) + u <- scale(x %*% v, center = FALSE, scale = d) + return(list(u = u, d = d, v = v)) + } + else { + junk <- svd(x, LINPACK = TRUE) + nc = min(ncol(junk$u), n.components) + return(list(u = junk$u[, 1:nc], d = junk$d[1:nc], v = junk$v[, + 1:nc])) + } } quantitative.seq.func <- function(xresamp, y) { - tt <- rep(0, dim(xresamp)[1]) - for (i in 1:dim(xresamp)[3]) { - y.ranked <- rank(y, ties.method = "random") - (dim(xresamp)[2] + - 1)/2 - tt <- tt + (xresamp[, , i] - (dim(xresamp)[2] + 1)/2) %*% - y.ranked - } - ns <- dim(xresamp)[2] - tt <- tt/(dim(xresamp)[3] * (ns^3 - ns)/12) - return(list(tt = as.vector(tt), numer = as.vector(tt), sd = rep(1, - length(tt)))) + tt <- rep(0, dim(xresamp)[1]) + for (i in 1:dim(xresamp)[3]) { + y.ranked <- rank(y, ties.method = "random") - (dim(xresamp)[2] + + 1)/2 + tt <- tt + (xresamp[, , i] - (dim(xresamp)[2] + 1)/2) %*% + y.ranked + } + ns <- dim(xresamp)[2] + tt <- tt/(dim(xresamp)[3] * (ns^3 - ns)/12) + return(list(tt = as.vector(tt), numer = as.vector(tt), sd = rep(1, + length(tt)))) } multiclass.seq.func <- function(xresamp, y) { - # number of classes and number of samples in each class - K <- max(y) - n.each <- rep(0, K) - for (k in 1 : K) - { - n.each[k] <- sum(y == k) - } - # the statistic - tt <- temp <- rep(0, dim(xresamp)[1]) - stand.contrasts <- matrix(0, dim(xresamp)[1], K) + # number of classes and number of samples in each class + K <- max(y) + n.each <- rep(0, K) + for (k in 1 : K) + { + n.each[k] <- sum(y == k) + } + # the statistic + tt <- temp <- rep(0, dim(xresamp)[1]) + stand.contrasts <- matrix(0, dim(xresamp)[1], K) - for (i in 1 : dim(xresamp)[3]) - { - for (k in 1 : K) - { - temp <- rowSums(xresamp[, y == k, i]) - tt <- tt + temp ^2 / n.each[k] - stand.contrasts[, k] <- stand.contrasts[, k] + temp - } - } - # finalize - nresamp <- dim(xresamp)[3] - ns <- dim(xresamp)[2] - tt <- tt / nresamp * 12 / ns / (ns + 1) - 3 * (ns + 1) - stand.contrasts <- stand.contrasts / nresamp - stand.contrasts <- scale(stand.contrasts, center=n.each * (ns + 1) / 2, - scale=sqrt(n.each * (ns - n.each) * (ns + 1) / 12)) - return(list(tt = tt, numer = tt, sd = rep(1, length(tt)), - stand.contrasts = stand.contrasts)) + for (i in 1 : dim(xresamp)[3]) + { + for (k in 1 : K) + { + temp <- rowSums(xresamp[, y == k, i]) + tt <- tt + temp ^2 / n.each[k] + stand.contrasts[, k] <- stand.contrasts[, k] + temp + } + } + # finalize + nresamp <- dim(xresamp)[3] + ns <- dim(xresamp)[2] + tt <- tt / nresamp * 12 / ns / (ns + 1) - 3 * (ns + 1) + stand.contrasts <- stand.contrasts / nresamp + stand.contrasts <- scale(stand.contrasts, center=n.each * (ns + 1) / 2, + scale=sqrt(n.each * (ns - n.each) * (ns + 1) / 12)) + return(list(tt = tt, numer = tt, sd = rep(1, length(tt)), + stand.contrasts = stand.contrasts)) } diff --git a/man/FinalPreprocessing.Rd b/man/FinalPreprocessing.Rd index e17db70..ba9bd9b 100644 --- a/man/FinalPreprocessing.Rd +++ b/man/FinalPreprocessing.Rd @@ -9,14 +9,16 @@ FinalPreprocessing( object, GeneFlitering = "NoiseF", export = FALSE, - quiet = FALSE + quiet = FALSE, + fileName = "filteredDataset" ) \S4method{FinalPreprocessing}{DISCBIO}( object, GeneFlitering = "NoiseF", export = FALSE, - quiet = FALSE + quiet = FALSE, + fileName = "filteredDataset" ) } \arguments{ @@ -29,6 +31,8 @@ FinalPreprocessing( excel file. Default is TRUE.} \item{quiet}{if `TRUE`, intermediary output is suppressed} + +\item{fileName}{File name for exporting (if `export = TRUE`)} } \value{ The DISCBIO-class object input with the FinalGeneList slot filled. diff --git a/vignettes/DIscBIO-vignette.Rmd b/vignettes/DIscBIO-vignette.Rmd index bdc89ed..7df17ee 100644 --- a/vignettes/DIscBIO-vignette.Rmd +++ b/vignettes/DIscBIO-vignette.Rmd @@ -137,7 +137,7 @@ plotGap(sc) ### Plotting gap statisticssc <- Clustexp(sc, clustnr=20,bootnr=50,m ```{r} sc <- comptSNE(sc,rseed=15555,quiet=TRUE) -cat("\t"," Cell-ID"," Cluster Number","\n") +message("\t"," Cell-ID"," Cluster Number") sc@cpart ``` @@ -188,7 +188,7 @@ if(RemovingOutliers==TRUE){ DataSet=DataSet[-Outliers] dim(DataSet) colnames(DataSet) - cat("Outlier cells were removed, now you need to start from the beginning") + message("Outlier cells were removed, now you need to start from the beginning") } ``` @@ -405,7 +405,7 @@ Model-based clustering assumes that the data are generated by a model and attemp ```{r, eval=FALSE} sc <- Exprmclust(sc,K=3,reduce=TRUE,quiet=TRUE) -cat("\t"," Cell-ID"," Cluster Number","\n") +message("\t"," Cell-ID"," Cluster Number") sc@MBclusters$clusterid ``` @@ -456,7 +456,7 @@ if(RemovingOutliers==TRUE){ DataSet=DataSet[-Outliers] dim(DataSet) colnames(DataSet) - cat("Outlier cells were removed, now you need to start from the beginning") + message("Outlier cells were removed, now you need to start from the beginning") } ``` From 15583374835c27b22c8f7709962bc0eb8fd8f766 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Tue, 21 Jul 2020 10:57:54 +0200 Subject: [PATCH 47/99] File renaming --- tests/testthat/{test.DEGfreeze.R => test-DEGfreeze.R} | 0 tests/testthat/{test.DIscBIO.IMP.R => test-DIscBIO.IMP.R} | 0 tests/testthat/{test.integration.R => test-integration.R} | 0 3 files changed, 0 insertions(+), 0 deletions(-) rename tests/testthat/{test.DEGfreeze.R => test-DEGfreeze.R} (100%) rename tests/testthat/{test.DIscBIO.IMP.R => test-DIscBIO.IMP.R} (100%) rename tests/testthat/{test.integration.R => test-integration.R} (100%) diff --git a/tests/testthat/test.DEGfreeze.R b/tests/testthat/test-DEGfreeze.R similarity index 100% rename from tests/testthat/test.DEGfreeze.R rename to tests/testthat/test-DEGfreeze.R diff --git a/tests/testthat/test.DIscBIO.IMP.R b/tests/testthat/test-DIscBIO.IMP.R similarity index 100% rename from tests/testthat/test.DIscBIO.IMP.R rename to tests/testthat/test-DIscBIO.IMP.R diff --git a/tests/testthat/test.integration.R b/tests/testthat/test-integration.R similarity index 100% rename from tests/testthat/test.integration.R rename to tests/testthat/test-integration.R From 4548e5d3101b3b4c56f61c656d2b9781b54a56e2 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Tue, 21 Jul 2020 11:11:49 +0200 Subject: [PATCH 48/99] Suppressed output on tests --- tests/testthat/test-DEGfreeze.R | 10 ++++++---- tests/testthat/test-DIscBIO.IMP.R | 18 +++++++++++------- 2 files changed, 17 insertions(+), 11 deletions(-) diff --git a/tests/testthat/test-DEGfreeze.R b/tests/testthat/test-DEGfreeze.R index 14599d5..3ce6b97 100644 --- a/tests/testthat/test-DEGfreeze.R +++ b/tests/testthat/test-DEGfreeze.R @@ -28,10 +28,12 @@ sc <- FinalPreprocessing(sc, GeneFlitering="ExpF", export=FALSE, quiet=TRUE) sc <- Clustexp( sc, clustnr=n_clust, quiet=TRUE, bootnr = 2, B.gap = 2, rseed=17000 ) -cdiff <- DEGanalysis( - sc, Clustering="K-means", K=n_clust, fdr=0.10, name="all_clusters", - export=FALSE, quiet=TRUE, plot=FALSE, nresamp=2, nperms=2 +cdiff <- suppressMessages( + DEGanalysis( + sc, Clustering="K-means", K=n_clust, fdr=0.10, name="all_clusters", + export=FALSE, quiet=TRUE, plot=FALSE, nresamp=2, nperms=2 + ) ) test_that("DEG analysis doesn't freeze", { expect_output(str(cdiff), "List of 2") -}) \ No newline at end of file +}) diff --git a/tests/testthat/test-DIscBIO.IMP.R b/tests/testthat/test-DIscBIO.IMP.R index 75ebf48..5d993f8 100644 --- a/tests/testthat/test-DIscBIO.IMP.R +++ b/tests/testthat/test-DIscBIO.IMP.R @@ -95,15 +95,19 @@ context("Differential Expression Analysis") cdiff1 <- KMClustDiffGenes(sc, K=1, fdr=.2, export=FALSE, quiet=TRUE) # differential expression analysis between all clusters -cdiff2 <- DEGanalysis( - sc, Clustering="K-means", K=2, fdr=.2, name="Name", export=FALSE, - quiet=TRUE, plot=FALSE, nperms=5, nresamp=2 +cdiff2 <- suppressMessages( + DEGanalysis( + sc, Clustering="K-means", K=2, fdr=.2, name="Name", export=FALSE, + quiet=TRUE, plot=FALSE, nperms=5, nresamp=2 + ) ) # differential expression analysis between two particular clusters. -cdiff3 <- DEGanalysis2clust( - sc, Clustering="K-means", K=2, fdr=.2, name="Name", First="CL1", - Second="CL2", export=FALSE, quiet=TRUE, plot=FALSE, nperms=5, nresamp=2 +cdiff3 <- suppressMessages( + DEGanalysis2clust( + sc, Clustering="K-means", K=2, fdr=.2, name="Name", First="CL1", + Second="CL2", export=FALSE, quiet=TRUE, plot=FALSE, nperms=5, nresamp=2 + ) ) test_that("DEGs are calculated", { @@ -147,4 +151,4 @@ test_that("Decision tree elements are defined", { expect_identical(j48dt_eval, c(TP = 7, FN = 4, FP = 5, TN = 5)) expect_s3_class(rpartDT, "rpart") expect_identical(rpartEVAL, c(TP = 5, FN = 6, FP = 10, TN = 0)) -}) \ No newline at end of file +}) From e9ca7cb5186afcd38ec00ce9d36d11cf2b3e248b Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Tue, 21 Jul 2020 11:23:36 +0200 Subject: [PATCH 49/99] Suppressed exporting in vignette --- vignettes/DIscBIO-vignette.Rmd | 21 +++++++++------------ 1 file changed, 9 insertions(+), 12 deletions(-) diff --git a/vignettes/DIscBIO-vignette.Rmd b/vignettes/DIscBIO-vignette.Rmd index 7df17ee..967ff1c 100644 --- a/vignettes/DIscBIO-vignette.Rmd +++ b/vignettes/DIscBIO-vignette.Rmd @@ -78,7 +78,7 @@ sc <- Normalizedata(sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, down #MinNumber #sc <- Normalizedata(sc, mintotal=1000, minexpr=MIinExp, minnumber=MinNumber, maxexpr=Inf, downsample=FALSE, dsn=1, rseed=17000) -sc <- FinalPreprocessing(sc,GeneFlitering="NoiseF",export=TRUE) ### The GeneFiltering can be either "NoiseF" or"ExpF" +sc <- FinalPreprocessing(sc,GeneFlitering="NoiseF") ### The GeneFiltering can be either "NoiseF" or"ExpF" ``` ### 1.2. Filtering and normalizing the raw data that does not include ERCCs @@ -100,7 +100,7 @@ if (OnlyExpressionFiltering==TRUE){ MinNumber <- round(length(DataSet[1,])/3) # To be expressed in at least one third of the cells. MinNumber sc <- Normalizedata(sc, mintotal=1000, minexpr=MIinExp, minnumber=MinNumber, maxexpr=Inf, downsample=FALSE, dsn=1, rseed=17000) #### In this case this function is used to filter out genes and cells. - sc <- FinalPreprocessing(sc,GeneFlitering="ExpF",export=TRUE) + sc <- FinalPreprocessing(sc,GeneFlitering="ExpF",export=FALSE) } ``` @@ -137,7 +137,6 @@ plotGap(sc) ### Plotting gap statisticssc <- Clustexp(sc, clustnr=20,bootnr=50,m ```{r} sc <- comptSNE(sc,rseed=15555,quiet=TRUE) -message("\t"," Cell-ID"," Cluster Number") sc@cpart ``` @@ -164,8 +163,8 @@ Here you visualize the K-means clusters using t-distributed stochastic neighbor ```{r} ############ Plotting K-means clusters plottSNE(sc) -plotKmeansLabelstSNE(sc) # To plot the the ID of the cells in eacj cluster -plotSymbolstSNE(sc,types=sub("(\\_\\d+)$","", names(sc@ndata))) # To plot the the ID of the cells in eacj cluster +plotKmeansLabelstSNE(sc) # To plot the the ID of the cells in each cluster +plotSymbolstSNE(sc,types=sub("(\\_\\d+)$","", names(sc@ndata))) # To plot the the ID of the cells in each cluster ``` ### Defining outlier cells based on K-means Clustering @@ -195,7 +194,7 @@ if(RemovingOutliers==TRUE){ #### 2.1.4. Cellular pseudo-time ordering based on k-means clusters ```{r} -sc <- KmeanOrder(sc,quiet=FALSE, export=TRUE) +sc <- KmeanOrder(sc,quiet=FALSE, export=FALSE) plotOrderKMtsne(sc) ``` @@ -212,8 +211,7 @@ KMclustheatmap(sc,hmethod="single", plot=TRUE) ```{r} -g='ENSG00000010244' #### Plotting the expression of MT-RNR2 -plotExptSNE(sc,g) +plotExptSNE(sc, "ENSG00000101346") ``` ## 3. Determining differentially expressed genes (DEGs) For K-means Clustering @@ -230,7 +228,7 @@ The user can define DEGs between all clusters generated by either K-means or mod ```{r degKM} ####### differential expression analysis between cluster 1 and cluster 3 of the Model-Based clustering using FDR of 0.05 -cdiff <- DEGanalysis2clust(sc,Clustering="K-means",K=2,fdr=0.05,name="Name",First="CL1",Second="CL2",export=TRUE,quiet=TRUE) +cdiff <- DEGanalysis2clust(sc,Clustering="K-means",K=2,fdr=0.05,name="Name",First="CL1",Second="CL2",export=FALSE,quiet=TRUE) ``` ```{r} @@ -246,7 +244,7 @@ head(cdiff[[2]]) # The second component ```{r} -cdiff <- DEGanalysis(sc,Clustering="K-means",K=2,fdr=0.05,name="all_clusters",export=TRUE,quiet=TRUE) ####### differential expression analysis between all clusters +cdiff <- DEGanalysis(sc,Clustering="K-means",K=2,fdr=0.05,name="all_clusters",export=FALSE,quiet=TRUE) ####### differential expression analysis between all clusters ``` ```{r} @@ -405,7 +403,6 @@ Model-based clustering assumes that the data are generated by a model and attemp ```{r, eval=FALSE} sc <- Exprmclust(sc,K=3,reduce=TRUE,quiet=TRUE) -message("\t"," Cell-ID"," Cluster Number") sc@MBclusters$clusterid ``` @@ -464,7 +461,7 @@ if(RemovingOutliers==TRUE){ ```{r, eval=FALSE} -sc <- MB_Order(sc,quiet=FALSE, export=TRUE) +sc <- MB_Order(sc,quiet=FALSE, export=FALSE) ``` #### 2.2.4.1 Plotting the pseudo-time ordering in a PCA plot From 75f1bbc4684d748901f66709688a4080adeb6796 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Tue, 21 Jul 2020 11:24:11 +0200 Subject: [PATCH 50/99] Removed MB from vignette --- vignettes/DIscBIO-vignette.Rmd | 109 --------------------------------- 1 file changed, 109 deletions(-) diff --git a/vignettes/DIscBIO-vignette.Rmd b/vignettes/DIscBIO-vignette.Rmd index 967ff1c..50a8cd2 100644 --- a/vignettes/DIscBIO-vignette.Rmd +++ b/vignettes/DIscBIO-vignette.Rmd @@ -393,112 +393,3 @@ GO_Biological_Process_2015 <- enriched[[3]][,c(1,2,3,9)] GEA <- rbind(KEGG_2013,GO_Molecular_Function_2015,GO_Biological_Process_2015) GEA ``` - -## 2.2. Model-Based clustering - -Model-based clustering assumes that the data are generated by a model and attempts to recover the original model from the data to define cellular clusters. The steps reproduced here are comparable to the ones on Section [2.1](#21-k-means-clustering). - -#### 2.2.1. Defining the Cells in the clusters generated by model-based clustering - - -```{r, eval=FALSE} -sc <- Exprmclust(sc,K=3,reduce=TRUE,quiet=TRUE) -sc@MBclusters$clusterid -``` - -#### 2.2.2. Cluster plotting using PCA and tSNE maps - -To visualize the detected clusters, two common dimensionality reduction tools are implemented: tSNE map and principal component analysis (PCA), which is a linear dimensionality reduction method that preserves the global structure and shows how the measurements themselves are related to each other. - - -```{r, eval=FALSE} -PlotmclustMB(sc) -PCAplotSymbols(sc) -``` - -```{r, eval=FALSE} -# Plotting the model-based clusters in tSNE maps -sc <- comptsneMB(sc,rseed=15555,quiet=TRUE) -plottsneMB(sc,K=3) -plotMBLabelstSNE(sc) -``` - -#### 2.2.3. Evaluating the stability and consistancy of the clusters - - -```{r, eval=FALSE} -# Silhouette of Model-based clusters -# you might have to run "par(mar=c(6,2,4,2))" first so plot looks better -plotsilhouetteMB(sc,K=3) -``` - -```{r, eval=FALSE} -sc <- Clustexp(sc,cln=3,quiet=TRUE) -Jaccard(sc,Clustering="MB", K=3, plot=TRUE) # Jaccard of Model based clusters -``` - -### Defining outlier cells based on Model-Based Clustering - - -```{r, eval=FALSE} -outlg <- round(length(sc@fdata[,1])/200) # The cell will be considered as an outlier if it has a minimum of 0.5% of the number of filtered genes as outlier genes. -Outliers <- FindOutliersMB(sc, K=3, outminc=5,outlg=outlg,probthr=.5*1e-3,thr=2**-(1:40),outdistquant=.75, plot=TRUE, quiet=TRUE) - -RemovingOutliers=FALSE -# RemovingOutliers=TRUE # Removing the defined outlier cells based on K-means Clustering - -if(RemovingOutliers==TRUE){ - names(Outliers)=NULL - Outliers - DataSet=DataSet[-Outliers] - dim(DataSet) - colnames(DataSet) - message("Outlier cells were removed, now you need to start from the beginning") -} -``` - -### 2.2.4. Cellular pseudo-time ordering based on Model-based clusters - - -```{r, eval=FALSE} -sc <- MB_Order(sc,quiet=FALSE, export=FALSE) -``` - -#### 2.2.4.1 Plotting the pseudo-time ordering in a PCA plot - - -```{r, eval=FALSE} -PlotMBorderPCA(sc) -``` - - -#### 2.2.4.2 Plotting the pseudo-time ordering in a tSNE map - - -```{r, eval=FALSE} -plotOrderMBtsne(sc) -``` - - -#### 2.2.4.3 Plotting the Model-based clusters in heatmap - - -```{r, eval=FALSE} -MBclustheatmap(sc,hmethod="single", plot=TRUE) -``` - -#### 2.2.4.4 Plotting the gene expression of a particular gene in a tSNE map - - -```{r, eval=FALSE} -g='ENSG00000010244' #### Plotting the expression of MT-RNR2 -plotexptsneMB(sc,g) -``` - -#### 2.2.4.5 Plotting the gene expression of a particular gene in a PCA plot - - -```{r, eval=FALSE} -g <- "ENSG00000010244" #### Plotting the expression of MT-RNR2 -PlotMBexpPCA(sc,g) -``` \ No newline at end of file From 1f191541733683237a1413ca97102a8204be466c Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Tue, 21 Jul 2020 11:38:02 +0200 Subject: [PATCH 51/99] Removed commented out code --- vignettes/DIscBIO-vignette.Rmd | 63 +++++++--------------------------- 1 file changed, 12 insertions(+), 51 deletions(-) diff --git a/vignettes/DIscBIO-vignette.Rmd b/vignettes/DIscBIO-vignette.Rmd index 50a8cd2..d25feaf 100644 --- a/vignettes/DIscBIO-vignette.Rmd +++ b/vignettes/DIscBIO-vignette.Rmd @@ -44,17 +44,18 @@ sc <- DISCBIO(DataSet) ### 1.1. Filtering and normalizing the raw data that includes ERCCs Filtering the raw data that includes ERCCs can be done by applying the “NoiseFiltering” function, which includes several parameters: - - object: the outcome of running the DISCBIO() function. - - percentile: A numeric value of the percentile. It is used to validate the ERCC spik-ins. Default is 0.8. - - CV: A numeric value of the coefficient of variation. It is used to validate the ERCC spik-ins. Default is 0.5. - - geneCol: Color of the genes that did not pass the filtration. - - FgeneCol: Color of the genes that passt the filtration. - - erccCol: Color of the ERCC spik-ins. - - Val: A logical vector that allows plotting only the validated ERCC spike-ins. Default is TRUE. If Val=FALSE will - plot all the ERCC spike-ins. - - plot: A logical vector that allows plotting the technical noise. Default is TRUE. - - export: A logical vector that allows writing the final gene list in excel file. Default is TRUE. - - quiet: if `TRUE`, suppresses printed output + + - object: the outcome of running the DISCBIO() function. + - percentile: A numeric value of the percentile. It is used to validate the ERCC spik-ins. Default is 0.8. + - CV: A numeric value of the coefficient of variation. It is used to validate the ERCC spik-ins. Default is 0.5. + - geneCol: Color of the genes that did not pass the filtration. + - FgeneCol: Color of the genes that passt the filtration. + - erccCol: Color of the ERCC spik-ins. + - Val: A logical vector that allows plotting only the validated ERCC spike-ins. Default is TRUE. If Val=FALSE will + plot all the ERCC spike-ins. + - plot: A logical vector that allows plotting the technical noise. Default is TRUE. + - export: A logical vector that allows writing the final gene list in excel file. Default is TRUE. + - quiet: if `TRUE`, suppresses printed output To normalize the raw sequencing reads the function Normalizedata() should be used, this function takes 8 parameters. - In case the user would like just to normalize the reads without any further gene filtering the parameters minexpr and minnumber should be set to 0. @@ -71,13 +72,6 @@ sc <- NoiseFiltering(sc,percentile=0.9, CV=0.2) #### Normalizing the reads without any further gene filtering sc <- Normalizedata(sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, dsn=1, rseed=17000) -#### Additional gene filtering step based on gene expression -#MIinExp <- mean(rowMeans(DataSet,na.rm=TRUE)) -#MIinExp -#MinNumber <- round(length(DataSet[1,])/3) # To be expressed in at least one third of the cells. -#MinNumber -#sc <- Normalizedata(sc, mintotal=1000, minexpr=MIinExp, minnumber=MinNumber, maxexpr=Inf, downsample=FALSE, dsn=1, rseed=17000) - sc <- FinalPreprocessing(sc,GeneFlitering="NoiseF") ### The GeneFiltering can be either "NoiseF" or"ExpF" ``` @@ -87,23 +81,6 @@ To normalize and filter the raw data that does not include ERCCs can be done by To Finalize the preprocessing the function FinalPreprocessing() should be implemented by setting the parameter "GeneFlitering" to ExpF. -**¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤ Running this cell will overwrite the previous cell ¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤** - - -```{r} -#OnlyExpressionFiltering=TRUE -OnlyExpressionFiltering=FALSE - -if (OnlyExpressionFiltering==TRUE){ - MIinExp <- mean(rowMeans(DataSet,na.rm=TRUE)) - MIinExp - MinNumber <- round(length(DataSet[1,])/3) # To be expressed in at least one third of the cells. - MinNumber - sc <- Normalizedata(sc, mintotal=1000, minexpr=MIinExp, minnumber=MinNumber, maxexpr=Inf, downsample=FALSE, dsn=1, rseed=17000) #### In this case this function is used to filter out genes and cells. - sc <- FinalPreprocessing(sc,GeneFlitering="ExpF",export=FALSE) -} -``` - ## 2. Cellular Clustering and Pseudo Time ordering Cellular clustering is performed according to the gene expression profiles to detect cellular sub-population with unique properties. After clustering, pseudo-temporal ordering is generated to indicate the cellular differentiation degree. @@ -177,18 +154,6 @@ In case the user decided to remove outlier cells, the user should set RemovingOu ```{r} outlg <- round(length(sc@fdata[,1])/200) # The cell will be considered as an outlier if it has a minimum of 0.5% of the number of filtered genes as outlier genes. Outliers <- FindOutliersKM(sc, K=2, outminc=5,outlg=outlg,probthr=.5*1e-3,thr=2**-(1:40),outdistquant=.75, plot=TRUE, quiet=FALSE) - -RemovingOutliers=FALSE -# RemovingOutliers=TRUE # Removing the defined outlier cells based on K-means Clustering - -if(RemovingOutliers==TRUE){ - names(Outliers)=NULL - Outliers - DataSet=DataSet[-Outliers] - dim(DataSet) - colnames(DataSet) - message("Outlier cells were removed, now you need to start from the beginning") -} ``` #### 2.1.4. Cellular pseudo-time ordering based on k-means clusters @@ -335,7 +300,6 @@ DEGs="All DEGs" FileName=paste0(DEGs) data <- cdiffBinomial[[1]] [,2] # DEGs gene list from Binomial analysis (taking only the firat 100 genes) -#data <- cdiff[[1]] [,2] # From the table of the differential expression analysis between all pairs of clusters ppi <- PPI(data,FileName) @@ -354,7 +318,6 @@ network <- Networking(data,FileName,plot_width=25, plot_height=10) ```{r} ############ Selecting the DEGs' table ############## DEGs=cdiffBinomial[[2]][1,4] # Up-regulated genes in cluster 1 (from the Binomial analysis) -#DEGs=cdiff[[2]][1,4] # Up-regulated genes in cluster 1 (from SAMseq) FileName=paste0(DEGs) data <- read.csv(file=paste0(DEGs),head=TRUE,sep=",") @@ -373,13 +336,11 @@ network <- Networking(data,FileName) ```{r} dbs <- listEnrichrDbs() head(dbs) -#print(dbs) ``` ```{r} ############ Selecting the DEGs' table ############## DEGs=cdiffBinomial[[2]][1,4] # Up-regulated genes in cluster 1 (from the Binomial analysis) -#DEGs=cdiff[[2]][1,4] # Up-regulated genes in cluster 1 (from SAMseq) data <- read.csv(file=paste0(DEGs),head=TRUE,sep=",") data <- as.character(data[,3]) From b7e5bfbe74826a136f48d74c8045194eb7561cf4 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Tue, 21 Jul 2020 11:48:24 +0200 Subject: [PATCH 52/99] Fixed example --- R/DIscBIO-generic-FindOutliersKM.R | 10 +++------- man/FindOutliersKM.Rd | 7 ++----- 2 files changed, 5 insertions(+), 12 deletions(-) diff --git a/R/DIscBIO-generic-FindOutliersKM.R b/R/DIscBIO-generic-FindOutliersKM.R index 7f49b94..338383c 100644 --- a/R/DIscBIO-generic-FindOutliersKM.R +++ b/R/DIscBIO-generic-FindOutliersKM.R @@ -24,11 +24,8 @@ #' of cells on each. #' @examples #' sc <- DISCBIO(valuesG1msRed) -#' sc <- Clustexp(sc, cln=3) # K-means clustering -#' Outliers <- FindOutliersKM( -#' sc, K=3, outminc=5, outlg=2, probthr=.5*1e-3, thr=2**-(1:40), -#' outdistquant=.75, plot = FALSE -#' ) +#' sc <- Clustexp(sc, cln=2) # K-means clustering +#' FindOutliersKM(sc, K=2) #' setGeneric( "FindOutliersKM", @@ -45,8 +42,7 @@ setMethod( "FindOutliersKM", signature = "DISCBIO", definition = function( - object, K, outminc, outlg, probthr, thr, outdistquant, plot = TRUE, - quiet = FALSE + object, K, outminc, outlg, probthr, thr, outdistquant, plot, quiet ) { if (length(object@kmeans$kpart) == 0) diff --git a/man/FindOutliersKM.Rd b/man/FindOutliersKM.Rd index f08cd13..63aa8a2 100644 --- a/man/FindOutliersKM.Rd +++ b/man/FindOutliersKM.Rd @@ -66,10 +66,7 @@ This functions performs the outlier identification } \examples{ sc <- DISCBIO(valuesG1msRed) -sc <- Clustexp(sc, cln=3) # K-means clustering -Outliers <- FindOutliersKM( - sc, K=3, outminc=5, outlg=2, probthr=.5*1e-3, thr=2**-(1:40), - outdistquant=.75, plot = FALSE -) +sc <- Clustexp(sc, cln=2) # K-means clustering +FindOutliersKM(sc, K=2) } From 4ef00633ef1d42990da97bf1783cb182e8f031ba Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Tue, 21 Jul 2020 12:00:39 +0200 Subject: [PATCH 53/99] Simplified DEG unit tests --- tests/testthat/test-DEGfreeze.R | 39 ------------------------------- tests/testthat/test-DIscBIO.IMP.R | 19 --------------- 2 files changed, 58 deletions(-) delete mode 100644 tests/testthat/test-DEGfreeze.R diff --git a/tests/testthat/test-DEGfreeze.R b/tests/testthat/test-DEGfreeze.R deleted file mode 100644 index 3ce6b97..0000000 --- a/tests/testthat/test-DEGfreeze.R +++ /dev/null @@ -1,39 +0,0 @@ -context("Stress-testing DEG analyses") -# ============================================================================== -# Determining contants -# ============================================================================== -n_genes <- 1e3 -n_clust <- 3 -manual_test <- FALSE -# ============================================================================== -# Defining datasets to be used -# ============================================================================== -if (manual_test) { - data(pan_indrop_matrix_8000cells_18556genes) - sc <- DISCBIO(pan_indrop_matrix_8000cells_18556genes[, seq_len(n_genes)]) -} else { - sc <- DISCBIO(valuesG1msRed) - sc <- NoiseFiltering(sc, plot=FALSE, export=FALSE, quiet=TRUE) -} - -# ============================================================================== -# Performing operations -# ============================================================================== -sc <- Normalizedata(sc) -sc <- FinalPreprocessing(sc, GeneFlitering="ExpF", export=FALSE, quiet=TRUE) - -# ------------------------------------------------------------------------------ -# DEG Analysis -# ------------------------------------------------------------------------------ -sc <- Clustexp( - sc, clustnr=n_clust, quiet=TRUE, bootnr = 2, B.gap = 2, rseed=17000 -) -cdiff <- suppressMessages( - DEGanalysis( - sc, Clustering="K-means", K=n_clust, fdr=0.10, name="all_clusters", - export=FALSE, quiet=TRUE, plot=FALSE, nresamp=2, nperms=2 - ) -) -test_that("DEG analysis doesn't freeze", { - expect_output(str(cdiff), "List of 2") -}) diff --git a/tests/testthat/test-DIscBIO.IMP.R b/tests/testthat/test-DIscBIO.IMP.R index 5d993f8..7faf07f 100644 --- a/tests/testthat/test-DIscBIO.IMP.R +++ b/tests/testthat/test-DIscBIO.IMP.R @@ -91,17 +91,6 @@ test_that("Outliers are the expected", { context("Differential Expression Analysis") -# Binomial differential expression analysis -cdiff1 <- KMClustDiffGenes(sc, K=1, fdr=.2, export=FALSE, quiet=TRUE) - -# differential expression analysis between all clusters -cdiff2 <- suppressMessages( - DEGanalysis( - sc, Clustering="K-means", K=2, fdr=.2, name="Name", export=FALSE, - quiet=TRUE, plot=FALSE, nperms=5, nresamp=2 - ) -) - # differential expression analysis between two particular clusters. cdiff3 <- suppressMessages( DEGanalysis2clust( @@ -111,14 +100,6 @@ cdiff3 <- suppressMessages( ) test_that("DEGs are calculated", { - expect_identical( - object = sapply(cdiff1, function(x) class(x)[1]), - expected = c("matrix", "data.frame") - ) - expect_identical( - object = sapply(cdiff2, function(x) class(x)[1]), - expected = c("matrix", "data.frame") - ) expect_equivalent( object = sapply(cdiff3, function(x) class(x)[1]), expected = c("matrix", "data.frame", "data.frame", "data.frame") From ce9d6b18e5efd0590425128725056a4c23ddc1cd Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Thu, 23 Jul 2020 10:44:30 +0200 Subject: [PATCH 54/99] Removed/simplified examples --- R/DIscBIO-classes.R | 7 ------- R/DIscBIO-generic-Clustexp.R | 3 --- R/DIscBIO-generic-Exprmclust.R | 10 ---------- R/DIscBIO-generic-FinalPreprocessing.R | 4 ---- R/DIscBIO-generic-FindOutliersMB.R | 14 -------------- R/DIscBIO-generic-NoiseFiltering.R | 4 ---- R/DIscBIO-generic-Normalizedata.R | 9 --------- R/DIscBIO-generic-comptSNE.R | 2 +- R/customConverters.R | 7 ------- R/datasets.R | 4 ---- 10 files changed, 1 insertion(+), 63 deletions(-) diff --git a/R/DIscBIO-classes.R b/R/DIscBIO-classes.R index 839f338..db2da5a 100644 --- a/R/DIscBIO-classes.R +++ b/R/DIscBIO-classes.R @@ -50,13 +50,6 @@ #' @exportClass DISCBIO #' #' @importClassesFrom SingleCellExperiment SingleCellExperiment -#' -#' @examples -#' class(valuesG1msRed) -#' G1_reclassified <- DISCBIO(valuesG1msRed) -#' class(G1_reclassified) -#' str(G1_reclassified, max.level=2) -#' identical(G1_reclassified@expdataAll, valuesG1msRed) #' @export DISCBIO <- setClass( Class = "DISCBIO", diff --git a/R/DIscBIO-generic-Clustexp.R b/R/DIscBIO-generic-Clustexp.R index e0aa94f..151bc1d 100644 --- a/R/DIscBIO-generic-Clustexp.R +++ b/R/DIscBIO-generic-Clustexp.R @@ -33,9 +33,6 @@ #' @importFrom fpc cluster.stats calinhara dudahart2 #' @importFrom methods is #' @return The DISCBIO-class object input with the cpart slot filled. -#' @examples -#' sc <- DISCBIO(valuesG1msRed) # changes signature of data -#' sc <- Clustexp(sc, cln=3) setGeneric("Clustexp", function(object, clustnr = 20, bootnr = 50, metric = "pearson", do.gap = TRUE, SE.method = "Tibs2001SEmax", SE.factor = .25, B.gap = 50, cln = 0, rseed = NULL, quiet = FALSE) diff --git a/R/DIscBIO-generic-Exprmclust.R b/R/DIscBIO-generic-Exprmclust.R index a7bbe9f..e381249 100644 --- a/R/DIscBIO-generic-Exprmclust.R +++ b/R/DIscBIO-generic-Exprmclust.R @@ -18,16 +18,6 @@ #' the MBclusters slot filled. If the `object` is a data frame, the function #' returns a named list containing the four objects that together correspond #' to the contents of the MBclusters slot. -#' @examples -#' sc <- DISCBIO(valuesG1msRed) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- Exprmclust(sc,K=2) -#' print(sc@MBclusters) setGeneric( name = "Exprmclust", def = function(object, diff --git a/R/DIscBIO-generic-FinalPreprocessing.R b/R/DIscBIO-generic-FinalPreprocessing.R index d7495fd..34178b2 100644 --- a/R/DIscBIO-generic-FinalPreprocessing.R +++ b/R/DIscBIO-generic-FinalPreprocessing.R @@ -8,10 +8,6 @@ #' @param quiet if `TRUE`, intermediary output is suppressed #' @param fileName File name for exporting (if `export = TRUE`) #' @return The DISCBIO-class object input with the FinalGeneList slot filled. -#' @examples -#' sc <- DISCBIO(valuesG1msRed) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) setGeneric( "FinalPreprocessing", function( diff --git a/R/DIscBIO-generic-FindOutliersMB.R b/R/DIscBIO-generic-FindOutliersMB.R index 98a58a2..478a7c1 100644 --- a/R/DIscBIO-generic-FindOutliersMB.R +++ b/R/DIscBIO-generic-FindOutliersMB.R @@ -22,20 +22,6 @@ #' @importFrom amap K #' @return A named vector of the genes containing outlying cells and the number #' of cells on each. -#' @examples -#' sc <- DISCBIO(valuesG1msRed) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE, plot=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- Exprmclust(sc, K=2) -#' FindOutliersMB( -#' sc, K=2, outminc=5, outlg=2, probthr=.5*1e-3, thr=2**-(1:40), -#' outdistquant=.75, plot = FALSE, quiet = TRUE -#' ) -#' setGeneric( name = "FindOutliersMB", def = function(object, diff --git a/R/DIscBIO-generic-NoiseFiltering.R b/R/DIscBIO-generic-NoiseFiltering.R index 84540eb..87744e9 100644 --- a/R/DIscBIO-generic-NoiseFiltering.R +++ b/R/DIscBIO-generic-NoiseFiltering.R @@ -26,10 +26,6 @@ #' @importFrom statmod glmgam.fit #' @note This function should be used only if the dataset has ERCC. #' @return The DISCBIO-class object input with the noiseF slot filled. -#' @examples -#' sc <- DISCBIO(valuesG1msRed) # changes signature of data -#' sd_filtered <- NoiseFiltering(sc, export=FALSE) -#' str(sd_filtered) setGeneric( name = "NoiseFiltering", def = function( diff --git a/R/DIscBIO-generic-Normalizedata.R b/R/DIscBIO-generic-Normalizedata.R index 8a7db05..ce2760c 100644 --- a/R/DIscBIO-generic-Normalizedata.R +++ b/R/DIscBIO-generic-Normalizedata.R @@ -23,15 +23,6 @@ #' results #' @include DIscBIO-classes.R #' @return The DISCBIO-class object input with the ndata and fdata slots filled. -#' @examples -#' sc <- DISCBIO(valuesG1msRed) # changes signature of data -#' -#' # In this case this function is used to normalize the reads -#' sc_normal <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' summary(sc_normal@fdata) setGeneric( "Normalizedata", function( diff --git a/R/DIscBIO-generic-comptSNE.R b/R/DIscBIO-generic-comptSNE.R index 045b060..758a460 100644 --- a/R/DIscBIO-generic-comptSNE.R +++ b/R/DIscBIO-generic-comptSNE.R @@ -13,7 +13,7 @@ #' @return The DISCBIO-class object input with the tsne slot filled. #' @examples #' sc <- DISCBIO(valuesG1msRed) # changes signature of data -#' sc <- Clustexp(sc, cln=3) # data must be clustered before plottin +#' sc <- Clustexp(sc, cln=2) # data must be clustered before plottin #' sc <- comptSNE(sc, max_iter=30) #' head(sc@tsne) setGeneric( diff --git a/R/customConverters.R b/R/customConverters.R index 7364e2c..b7d9b26 100644 --- a/R/customConverters.R +++ b/R/customConverters.R @@ -80,13 +80,6 @@ customConvertFeats <- function(x, verbose = TRUE) { #' #' @export #' -#' @examples -#' g1_sce <- SingleCellExperiment::SingleCellExperiment( -#' list(counts=as.matrix(valuesG1msRed)) -#' ) -#' g1_disc <- as.DISCBIO(g1_sce) -#' class(g1_disc) -#' as.DISCBIO <- function(x, ...) { if ("Seurat" %in% class(x)) { # Get Arguments and parse out what we want diff --git a/R/datasets.R b/R/datasets.R index fc26474..3a6b4c6 100644 --- a/R/datasets.R +++ b/R/datasets.R @@ -18,10 +18,6 @@ NULL #' @source Data were imported, modified, and formatted from the #' Mus.musculus (ver 1.3.1) and the Homo.sapiens (ver 1.3.1) #' BioConductor libraries. -#' -#' @examples -#' data(HumanMouseGeneIds) -#' print(HumanMouseGeneIds[1:6,]) #' @name HumanMouseGeneIds #' @docType data NULL From 56382be7351642dc475c3c78d63f8bc56d2543ef Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Thu, 23 Jul 2020 10:44:58 +0200 Subject: [PATCH 55/99] Changed K in vignette --- vignettes/DIscBIO-vignette.Rmd | 4 ++-- 1 file changed, 2 insertions(+), 2 deletions(-) diff --git a/vignettes/DIscBIO-vignette.Rmd b/vignettes/DIscBIO-vignette.Rmd index d25feaf..bfd769a 100644 --- a/vignettes/DIscBIO-vignette.Rmd +++ b/vignettes/DIscBIO-vignette.Rmd @@ -106,7 +106,7 @@ The Clustexp() functions has several parameters: ```{r} sc <- Clustexp(sc,cln=2,quiet=TRUE) #### K-means clustering to get three clusters -plotGap(sc) ### Plotting gap statisticssc <- Clustexp(sc, clustnr=20,bootnr=50,metric="pearson",do.gap=TRUE,SE.method="Tibs2001SEmax",SE.factor=.25,B.gap=50,cln=K,rseed=17000) +plotGap(sc) ### Plotting gap statisticssc <- Clustexp(sc, clustnr=20,bootnr=50,metric="pearson",do.gap=TRUE,SE.method="Tibs2001SEmax",SE.factor=.25,B.gap=50,cln=2,rseed=17000) ``` #### 2.1.1. Defining the Cells in the clusters generated by k-means clustering @@ -256,7 +256,7 @@ Decision trees are one of the most efficient classification techniques in biomar sigDEG <- cdiffBinomial[[1]] # DEGs gene list from Binomial analysis First="CL1" Second="CL2" -DATAforDT <- ClassVectoringDT(sc,Clustering="K-means",K=3,First=First,Second=Second,sigDEG) +DATAforDT <- ClassVectoringDT(sc,Clustering="K-means",K=2,First=First,Second=Second,sigDEG) ``` ### 4.1. J48 Decision Tree From 00479c99ae1d21b160eea82145b813fd92b84e4e Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Thu, 23 Jul 2020 10:45:06 +0200 Subject: [PATCH 56/99] Updated documentation --- man/Clustexp.Rd | 4 ---- man/DISCBIO.Rd | 7 ------- man/Exprmclust.Rd | 11 ----------- man/FinalPreprocessing.Rd | 5 ----- man/FindOutliersMB.Rd | 15 --------------- man/HumanMouseGeneIds.Rd | 4 ---- man/NoiseFiltering.Rd | 5 ----- man/Normalizedata.Rd | 10 ---------- man/as.DISCBIO.Rd | 8 -------- man/comptSNE.Rd | 2 +- tests/testthat/test-integration.R | 2 +- 11 files changed, 2 insertions(+), 71 deletions(-) diff --git a/man/Clustexp.Rd b/man/Clustexp.Rd index a4350f5..7711797 100644 --- a/man/Clustexp.Rd +++ b/man/Clustexp.Rd @@ -76,7 +76,3 @@ The DISCBIO-class object input with the cpart slot filled. This functions performs the initial clustering of the RaceID algorithm. } -\examples{ -sc <- DISCBIO(valuesG1msRed) # changes signature of data -sc <- Clustexp(sc, cln=3) -} diff --git a/man/DISCBIO.Rd b/man/DISCBIO.Rd index 73ba2fe..261b1b2 100644 --- a/man/DISCBIO.Rd +++ b/man/DISCBIO.Rd @@ -82,10 +82,3 @@ noise filtering.} from running the noise filtering or/and the expression filtering.} }} -\examples{ -class(valuesG1msRed) -G1_reclassified <- DISCBIO(valuesG1msRed) -class(G1_reclassified) -str(G1_reclassified, max.level=2) -identical(G1_reclassified@expdataAll, valuesG1msRed) -} diff --git a/man/Exprmclust.Rd b/man/Exprmclust.Rd index 87b4f59..82d42b7 100644 --- a/man/Exprmclust.Rd +++ b/man/Exprmclust.Rd @@ -60,14 +60,3 @@ this function first uses principal component analysis (PCA) to reduce dimensionality of original data. It then performs model-based clustering on the transformed expression values. } -\examples{ -sc <- DISCBIO(valuesG1msRed) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -sc <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, - dsn=1, rseed=17000 -) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -sc <- Exprmclust(sc,K=2) -print(sc@MBclusters) -} diff --git a/man/FinalPreprocessing.Rd b/man/FinalPreprocessing.Rd index ba9bd9b..e6506c8 100644 --- a/man/FinalPreprocessing.Rd +++ b/man/FinalPreprocessing.Rd @@ -40,8 +40,3 @@ The DISCBIO-class object input with the FinalGeneList slot filled. \description{ This function generates the final filtered normalized dataset. } -\examples{ -sc <- DISCBIO(valuesG1msRed) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -} diff --git a/man/FindOutliersMB.Rd b/man/FindOutliersMB.Rd index 531abf1..82173b3 100644 --- a/man/FindOutliersMB.Rd +++ b/man/FindOutliersMB.Rd @@ -64,18 +64,3 @@ A named vector of the genes containing outlying cells and the number \description{ This functions performs the outlier identification } -\examples{ -sc <- DISCBIO(valuesG1msRed) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE, plot=FALSE) -sc <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, - dsn=1, rseed=17000 -) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -sc <- Exprmclust(sc, K=2) -FindOutliersMB( - sc, K=2, outminc=5, outlg=2, probthr=.5*1e-3, thr=2**-(1:40), - outdistquant=.75, plot = FALSE, quiet = TRUE -) - -} diff --git a/man/HumanMouseGeneIds.Rd b/man/HumanMouseGeneIds.Rd index 75d70a2..eb9b231 100644 --- a/man/HumanMouseGeneIds.Rd +++ b/man/HumanMouseGeneIds.Rd @@ -13,7 +13,3 @@ BioConductor libraries. Data.frame including ENTREZID, SYMBOL, and ENSEMBL gene identifiers of human and mouse genes. } -\examples{ -data(HumanMouseGeneIds) -print(HumanMouseGeneIds[1:6,]) -} diff --git a/man/NoiseFiltering.Rd b/man/NoiseFiltering.Rd index 77db8fb..b043910 100644 --- a/man/NoiseFiltering.Rd +++ b/man/NoiseFiltering.Rd @@ -74,8 +74,3 @@ Given a matrix or data frame of count data, this function \note{ This function should be used only if the dataset has ERCC. } -\examples{ -sc <- DISCBIO(valuesG1msRed) # changes signature of data -sd_filtered <- NoiseFiltering(sc, export=FALSE) -str(sd_filtered) -} diff --git a/man/Normalizedata.Rd b/man/Normalizedata.Rd index a9851f0..3c7e8dc 100644 --- a/man/Normalizedata.Rd +++ b/man/Normalizedata.Rd @@ -63,13 +63,3 @@ The DISCBIO-class object input with the ndata and fdata slots filled. This function allows filtering of genes and cells to be used in the downstream analysis. } -\examples{ -sc <- DISCBIO(valuesG1msRed) # changes signature of data - -# In this case this function is used to normalize the reads -sc_normal <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, - dsn=1, rseed=17000 -) -summary(sc_normal@fdata) -} diff --git a/man/as.DISCBIO.Rd b/man/as.DISCBIO.Rd index 85ce67a..11ad971 100644 --- a/man/as.DISCBIO.Rd +++ b/man/as.DISCBIO.Rd @@ -23,11 +23,3 @@ Additional parameters to pass to `list` include, if x is a Seurat object, "assay", which is a string indicating the assay slot used to obtain data from (defaults to 'RNA') } -\examples{ -g1_sce <- SingleCellExperiment::SingleCellExperiment( - list(counts=as.matrix(valuesG1msRed)) -) -g1_disc <- as.DISCBIO(g1_sce) -class(g1_disc) - -} diff --git a/man/comptSNE.Rd b/man/comptSNE.Rd index b832d47..5eee426 100644 --- a/man/comptSNE.Rd +++ b/man/comptSNE.Rd @@ -46,7 +46,7 @@ This function is used to compute the t-Distributed Stochastic } \examples{ sc <- DISCBIO(valuesG1msRed) # changes signature of data -sc <- Clustexp(sc, cln=3) # data must be clustered before plottin +sc <- Clustexp(sc, cln=2) # data must be clustered before plottin sc <- comptSNE(sc, max_iter=30) head(sc@tsne) } diff --git a/tests/testthat/test-integration.R b/tests/testthat/test-integration.R index d4a9c55..cad4f06 100644 --- a/tests/testthat/test-integration.R +++ b/tests/testthat/test-integration.R @@ -5,7 +5,7 @@ context("Converting other formats to DISCBIO") # ------------------------------------------------------------------------------ # Setting up datasets # ------------------------------------------------------------------------------ -pmbc_seurat <- Seurat::pbmc_small # TODO: replace with library(Seurat); data(...) to better pass r-devel? +pmbc_seurat <- Seurat::pbmc_small pmbc_sce <- Seurat::as.SingleCellExperiment(pmbc_seurat) g1_sce <- SingleCellExperiment::SingleCellExperiment( list(counts=as.matrix(valuesG1msRed)) From 7e90007e6991a5e82af47f44d96332d26cef95bf Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Wed, 29 Jul 2020 13:03:36 +0200 Subject: [PATCH 57/99] Removed news file --- NEWS.md | 2 -- 1 file changed, 2 deletions(-) delete mode 100644 NEWS.md diff --git a/NEWS.md b/NEWS.md deleted file mode 100644 index 6a3face..0000000 --- a/NEWS.md +++ /dev/null @@ -1,2 +0,0 @@ -Changes in version 0.99.0 (2020-02-17) -+ Submitted to Bioconductor From c35f270d49aa5b2b3bc0491bfd211c02da7dd350 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Wed, 29 Jul 2020 13:05:33 +0200 Subject: [PATCH 58/99] Updated documentation --- DESCRIPTION | 2 +- TODO.md | 2 +- 2 files changed, 2 insertions(+), 2 deletions(-) diff --git a/DESCRIPTION b/DESCRIPTION index 373e866..34c920f 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -1,5 +1,5 @@ Package: DIscBIO -Date: 2020-06-16 +Date: 2020-07-29 Title: A User-Friendly Pipeline for Biomarker Discovery in Single-Cell Transcriptomics Version: 1.0.0 Authors@R: diff --git a/TODO.md b/TODO.md index d23b309..431d80d 100644 --- a/TODO.md +++ b/TODO.md @@ -14,5 +14,5 @@ Ideally, the non-critical tasks below would also be addressed before release: - [x] Reduce data folder size - [ ] Standardize capitalization of function names and arguments - [x] Remove unessential variable assignments from [test.reproduceNotebook.R](tests/testthat/test.reproduceNotebook.R) -- [ ] Shorten examples with CPU or elapsed time > 5s +- [x] Shorten examples with CPU or elapsed time > 5s - [x] Reduce unit test length From 49235bb4b8d930c6122a1c9ac5aa322340409116 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Wed, 29 Jul 2020 13:06:20 +0200 Subject: [PATCH 59/99] Removed examples --- R/DIscBIO-generic-FindOutliersKM.R | 4 ---- R/DIscBIO-generic-KMClustDiffGenes.R | 5 +---- R/DIscBIO-generic-PCAplotSymbols.R | 11 +---------- R/DIscBIO-generic-comptSNE.R | 6 +----- R/DIscBIO-generic-comptsneMB.R | 12 +----------- R/customConverters.R | 6 ------ man/DISCBIO2SingleCellExperiment.Rd | 7 ------- man/FindOutliersKM.Rd | 6 ------ man/KMClustDiffGenes.Rd | 5 ----- man/PCAplotSymbols.Rd | 11 ----------- man/comptSNE.Rd | 6 ------ man/comptsneMB.Rd | 12 ------------ 12 files changed, 4 insertions(+), 87 deletions(-) diff --git a/R/DIscBIO-generic-FindOutliersKM.R b/R/DIscBIO-generic-FindOutliersKM.R index 338383c..40150b0 100644 --- a/R/DIscBIO-generic-FindOutliersKM.R +++ b/R/DIscBIO-generic-FindOutliersKM.R @@ -22,10 +22,6 @@ #' @importFrom amap K #' @return A named vector of the genes containing outlying cells and the number #' of cells on each. -#' @examples -#' sc <- DISCBIO(valuesG1msRed) -#' sc <- Clustexp(sc, cln=2) # K-means clustering -#' FindOutliersKM(sc, K=2) #' setGeneric( "FindOutliersKM", diff --git a/R/DIscBIO-generic-KMClustDiffGenes.R b/R/DIscBIO-generic-KMClustDiffGenes.R index 5d043cd..d6b1eda 100644 --- a/R/DIscBIO-generic-KMClustDiffGenes.R +++ b/R/DIscBIO-generic-KMClustDiffGenes.R @@ -16,10 +16,7 @@ #' @rdname KMClustDiffGenes #' @return A list containing two tables. #' @export -#' @examples -#' sc <- DISCBIO(valuesG1msRed) -#' sc <- Clustexp(sc, cln=3, quiet=TRUE) # K-means clustering -#' KMClustDiffGenes(sc, K=3, fdr=.3, export=FALSE) +#' setGeneric( "KMClustDiffGenes", function( diff --git a/R/DIscBIO-generic-PCAplotSymbols.R b/R/DIscBIO-generic-PCAplotSymbols.R index 391c9d3..0de0caf 100644 --- a/R/DIscBIO-generic-PCAplotSymbols.R +++ b/R/DIscBIO-generic-PCAplotSymbols.R @@ -6,16 +6,7 @@ #' @importFrom grDevices rainbow #' @importFrom graphics legend #' @return Plot of the Principal Components -#' @examples -#' sc <- DISCBIO(valuesG1msRed) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- Exprmclust(sc, K=2) -#' PCAplotSymbols(sc) +#' setGeneric("PCAplotSymbols", function(object, types = NULL) standardGeneric("PCAplotSymbols")) diff --git a/R/DIscBIO-generic-comptSNE.R b/R/DIscBIO-generic-comptSNE.R index 758a460..f65fabb 100644 --- a/R/DIscBIO-generic-comptSNE.R +++ b/R/DIscBIO-generic-comptSNE.R @@ -11,11 +11,7 @@ #' @importFrom tsne tsne #' @importFrom stats as.dist cor #' @return The DISCBIO-class object input with the tsne slot filled. -#' @examples -#' sc <- DISCBIO(valuesG1msRed) # changes signature of data -#' sc <- Clustexp(sc, cln=2) # data must be clustered before plottin -#' sc <- comptSNE(sc, max_iter=30) -#' head(sc@tsne) +#' setGeneric( name = "comptSNE", def = function(object, rseed = NULL, max_iter = 5000, epoch = 500, diff --git a/R/DIscBIO-generic-comptsneMB.R b/R/DIscBIO-generic-comptsneMB.R index 529b6e0..e6bca24 100644 --- a/R/DIscBIO-generic-comptsneMB.R +++ b/R/DIscBIO-generic-comptsneMB.R @@ -10,17 +10,7 @@ #' @importFrom tsne tsne #' @importFrom stats as.dist cor #' @return The DISCBIO-class object input with the MBtsne slot filled. -#' @examples -#' sc <- DISCBIO(valuesG1msRed) -#' sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE, plot=FALSE) -#' sc <- Normalizedata( -#' sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, -#' dsn=1, rseed=17000 -#' ) -#' sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -#' sc <- Exprmclust(sc, K=2) -#' sc <- comptsneMB(sc, rseed=15555, max_iter = 1000) -#' print(sc@MBtsne) +#' setGeneric( name = "comptsneMB", def = function(object, rseed = NULL, max_iter = 5000, epoch = 500, diff --git a/R/customConverters.R b/R/customConverters.R index b7d9b26..fa36afe 100644 --- a/R/customConverters.R +++ b/R/customConverters.R @@ -149,12 +149,6 @@ as.DISCBIO <- function(x, ...) { #' #' @export #' -#' @examples -#' g1_disc <- DISCBIO(valuesG1msRed) -#' class(g1_disc) -#' g1_sce <- DISCBIO2SingleCellExperiment(g1_disc) -#' class(g1_sce) -#' DISCBIO2SingleCellExperiment <- function(x) { return(x@SingleCellExperiment) } diff --git a/man/DISCBIO2SingleCellExperiment.Rd b/man/DISCBIO2SingleCellExperiment.Rd index 0d6ba72..adad853 100644 --- a/man/DISCBIO2SingleCellExperiment.Rd +++ b/man/DISCBIO2SingleCellExperiment.Rd @@ -16,10 +16,3 @@ a SingleCellExperiment-class object Extract the SingleCellExperiment input data from the corresponding input slot in a DISCBIO-class object } -\examples{ -g1_disc <- DISCBIO(valuesG1msRed) -class(g1_disc) -g1_sce <- DISCBIO2SingleCellExperiment(g1_disc) -class(g1_sce) - -} diff --git a/man/FindOutliersKM.Rd b/man/FindOutliersKM.Rd index 63aa8a2..fdc8124 100644 --- a/man/FindOutliersKM.Rd +++ b/man/FindOutliersKM.Rd @@ -64,9 +64,3 @@ A named vector of the genes containing outlying cells and the number \description{ This functions performs the outlier identification } -\examples{ -sc <- DISCBIO(valuesG1msRed) -sc <- Clustexp(sc, cln=2) # K-means clustering -FindOutliersKM(sc, K=2) - -} diff --git a/man/KMClustDiffGenes.Rd b/man/KMClustDiffGenes.Rd index eef6539..bd77c73 100644 --- a/man/KMClustDiffGenes.Rd +++ b/man/KMClustDiffGenes.Rd @@ -59,8 +59,3 @@ A list containing two tables. \description{ description } -\examples{ -sc <- DISCBIO(valuesG1msRed) -sc <- Clustexp(sc, cln=3, quiet=TRUE) # K-means clustering -KMClustDiffGenes(sc, K=3, fdr=.3, export=FALSE) -} diff --git a/man/PCAplotSymbols.Rd b/man/PCAplotSymbols.Rd index 9ba0d4a..f83a6e4 100644 --- a/man/PCAplotSymbols.Rd +++ b/man/PCAplotSymbols.Rd @@ -21,14 +21,3 @@ Plot of the Principal Components \description{ Generates a plot of grouped PCA components } -\examples{ -sc <- DISCBIO(valuesG1msRed) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE) -sc <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, - dsn=1, rseed=17000 -) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -sc <- Exprmclust(sc, K=2) -PCAplotSymbols(sc) -} diff --git a/man/comptSNE.Rd b/man/comptSNE.Rd index 5eee426..3d80f40 100644 --- a/man/comptSNE.Rd +++ b/man/comptSNE.Rd @@ -44,9 +44,3 @@ The DISCBIO-class object input with the tsne slot filled. This function is used to compute the t-Distributed Stochastic Neighbor Embedding (t-SNE). } -\examples{ -sc <- DISCBIO(valuesG1msRed) # changes signature of data -sc <- Clustexp(sc, cln=2) # data must be clustered before plottin -sc <- comptSNE(sc, max_iter=30) -head(sc@tsne) -} diff --git a/man/comptsneMB.Rd b/man/comptsneMB.Rd index 8cd1938..ec623b3 100644 --- a/man/comptsneMB.Rd +++ b/man/comptsneMB.Rd @@ -43,15 +43,3 @@ The DISCBIO-class object input with the MBtsne slot filled. This function is used to compute the t-Distributed Stochastic Neighbor Embedding (t-SNE). } -\examples{ -sc <- DISCBIO(valuesG1msRed) -sc <- NoiseFiltering(sc, percentile=0.9, CV=0.2, export=FALSE, plot=FALSE) -sc <- Normalizedata( - sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, - dsn=1, rseed=17000 -) -sc <- FinalPreprocessing(sc, GeneFlitering="NoiseF", export=FALSE) -sc <- Exprmclust(sc, K=2) -sc <- comptsneMB(sc, rseed=15555, max_iter = 1000) -print(sc@MBtsne) -} From adf3f04facba5b12524139fc8d60efed3d07151e Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Wed, 29 Jul 2020 13:07:10 +0200 Subject: [PATCH 60/99] Simplified IMP tests --- tests/testthat/test-DIscBIO.IMP.R | 3 --- 1 file changed, 3 deletions(-) diff --git a/tests/testthat/test-DIscBIO.IMP.R b/tests/testthat/test-DIscBIO.IMP.R index 7faf07f..bb5a492 100644 --- a/tests/testthat/test-DIscBIO.IMP.R +++ b/tests/testthat/test-DIscBIO.IMP.R @@ -118,18 +118,15 @@ j48dt <- J48DT(DATAforDT, quiet = TRUE, plot = FALSE) j48dt_eval <- J48DTeval( DATAforDT, num.folds=10, First="CL1", Second="CL2", quiet=TRUE ) -rpartDT <- RpartDT(DATAforDT, quiet = TRUE, plot = FALSE) rpartEVAL <- RpartEVAL( DATAforDT, num.folds=10, First="CL1", Second="CL2", quiet = TRUE ) test_that("Decision tree elements are defined", { expect_output(str(DATAforDT), "5 obs. of 21 variables") - expect_s3_class(j48dt, "J48") expect_identical( attributes(j48dt)$class, c("J48", "Weka_tree", "Weka_classifier") ) expect_identical(j48dt_eval, c(TP = 7, FN = 4, FP = 5, TN = 5)) - expect_s3_class(rpartDT, "rpart") expect_identical(rpartEVAL, c(TP = 5, FN = 6, FP = 10, TN = 0)) }) From 722d86eca0d2fe8f5d6ca239d48c12cc73008e89 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Wed, 29 Jul 2020 15:20:09 +0200 Subject: [PATCH 61/99] Removed dependencies --- DESCRIPTION | 9 +- NAMESPACE | 12 --- R/DIscBIO-generic-ClassVectoringDT.R | 1 - R/DIscBIO-generic-Clustexp.R | 16 +--- R/DIscBIO-generic-FindOutliersKM.R | 1 - R/DIscBIO-generic-FindOutliersMB.R | 1 - R/DIscBIO-generic-KMClustDiffGenes.R | 13 ++- R/DIscBIO-generic-MBClustDiffGenes.R | 12 +-- R/DIscBIO-generic-NoiseFiltering.R | 5 +- R/J48DT.R | 18 +--- R/Jaccard.R | 1 + R/VolcanoPlot.R | 3 +- R/retrieveBiomart.R | 123 --------------------------- R/samr-adapted.R | 12 +-- man/retrieveBiomart.Rd | 28 ------ 15 files changed, 31 insertions(+), 224 deletions(-) delete mode 100644 R/retrieveBiomart.R delete mode 100644 man/retrieveBiomart.Rd diff --git a/DESCRIPTION b/DESCRIPTION index 34c920f..db8e912 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -37,10 +37,10 @@ Description: An open, multi-algorithmic pipeline for easy, fast and efficient . License: MIT + file LICENSE Encoding: UTF-8 -Imports: methods, TSCAN, boot, httr, mclust, statmod, biomaRt, samr, igraph, - RWeka, partykit, grid, philentropy, NetIndices, png, matrixStats, grDevices, - readr, RColorBrewer, ggplot2, rpart, fpc, cluster, rpart.plot, amap, dplyr, - tsne, AnnotationDbi, org.Hs.eg.db, calibrate, graphics, stats, utils, impute +Imports: methods, TSCAN, boot, httr, mclust, statmod, samr, igraph, + RWeka, philentropy, NetIndices, png, grDevices, + readr, RColorBrewer, ggplot2, rpart, fpc, cluster, rpart.plot, + tsne, AnnotationDbi, org.Hs.eg.db, graphics, stats, utils, impute Depends: R (>= 4.0), SingleCellExperiment Suggests: @@ -104,5 +104,4 @@ Collate: 'prepExampleDataset.R' 'reformatSiggenes.R' 'replaceDecimals.R' - 'retrieveBiomart.R' 'samr-adapted.R' diff --git a/NAMESPACE b/NAMESPACE index 33526ff..b0da0b0 100644 --- a/NAMESPACE +++ b/NAMESPACE @@ -58,18 +58,10 @@ importFrom(NetIndices,GenInd) importFrom(RColorBrewer,brewer.pal) importFrom(RWeka,J48) importFrom(TSCAN,TSCANorder) -importFrom(amap,K) -importFrom(amap,Kmeans) -importFrom(biomaRt,getBM) -importFrom(biomaRt,useDataset) -importFrom(biomaRt,useEnsembl) -importFrom(biomaRt,useMart) importFrom(boot,boot) -importFrom(calibrate,textxy) importFrom(cluster,clusGap) importFrom(cluster,maxSE) importFrom(cluster,silhouette) -importFrom(dplyr,summarize) importFrom(fpc,calinhara) importFrom(fpc,cluster.stats) importFrom(fpc,clusterboot) @@ -109,7 +101,6 @@ importFrom(graphics,points) importFrom(graphics,rasterImage) importFrom(graphics,text) importFrom(graphics,title) -importFrom(grid,gpar) importFrom(httr,GET) importFrom(httr,content) importFrom(httr,status_code) @@ -128,14 +119,11 @@ importFrom(igraph,graph.data.frame) importFrom(igraph,mean_distance) importFrom(igraph,minimum.spanning.tree) importFrom(impute,impute.knn) -importFrom(matrixStats,rowMedians) -importFrom(matrixStats,rowVars) importFrom(mclust,Mclust) importFrom(mclust,mclustBIC) importFrom(methods,is) importFrom(methods,new) importFrom(methods,validObject) -importFrom(partykit,as.party) importFrom(philentropy,distance) importFrom(png,readPNG) importFrom(readr,read_tsv) diff --git a/R/DIscBIO-generic-ClassVectoringDT.R b/R/DIscBIO-generic-ClassVectoringDT.R index 9e1f2a1..8984af7 100644 --- a/R/DIscBIO-generic-ClassVectoringDT.R +++ b/R/DIscBIO-generic-ClassVectoringDT.R @@ -12,7 +12,6 @@ #' @param sigDEG A data frame of the differentially expressed genes (DEGs) #' generated by running "DEGanalysis()" or "DEGanalysisM()". #' @param quiet If `TRUE`, suppresses intermediary output -#' @importFrom biomaRt useDataset useMart getBM #' @return A data frame. setGeneric( "ClassVectoringDT", diff --git a/R/DIscBIO-generic-Clustexp.R b/R/DIscBIO-generic-Clustexp.R index 151bc1d..b4227e2 100644 --- a/R/DIscBIO-generic-Clustexp.R +++ b/R/DIscBIO-generic-Clustexp.R @@ -27,10 +27,8 @@ #' @param quiet if `TRUE`, intermediate output is suppressed #' @importFrom stats as.dist cor kmeans #' @importFrom cluster clusGap maxSE -#' @importFrom fpc clusterboot -#' @importFrom amap Kmeans +#' @importFrom fpc clusterboot cluster.stats calinhara dudahart2 #' @importFrom graphics pairs -#' @importFrom fpc cluster.stats calinhara dudahart2 #' @importFrom methods is #' @return The DISCBIO-class object input with the cpart slot filled. setGeneric("Clustexp", function(object, clustnr = 20, bootnr = 50, @@ -204,16 +202,8 @@ setMethod( opar <- options(show.error.messages = FALSE) on.exit(options(opar)) repeat { - kmm <- try(Kmeans( - data, - k, - iter.max = iter.max, - method = method, - ... - )) - if (!is(kmm, - "try-error")) - break + kmm <- try(kmeans(data, k)) + if (!is(kmm, "try-error")) break } opar <- options(show.error.messages = TRUE) on.exit(options(opar)) diff --git a/R/DIscBIO-generic-FindOutliersKM.R b/R/DIscBIO-generic-FindOutliersKM.R index 40150b0..aa11009 100644 --- a/R/DIscBIO-generic-FindOutliersKM.R +++ b/R/DIscBIO-generic-FindOutliersKM.R @@ -19,7 +19,6 @@ #' @param plot if `TRUE`, produces a plot of -log10prob per K #' @param quiet if `TRUE`, intermediary output is suppressed #' @importFrom stats coef pnbinom -#' @importFrom amap K #' @return A named vector of the genes containing outlying cells and the number #' of cells on each. #' diff --git a/R/DIscBIO-generic-FindOutliersMB.R b/R/DIscBIO-generic-FindOutliersMB.R index 478a7c1..f3116f7 100644 --- a/R/DIscBIO-generic-FindOutliersMB.R +++ b/R/DIscBIO-generic-FindOutliersMB.R @@ -19,7 +19,6 @@ #' @param plot if `TRUE`, produces a plot of -log10prob per K #' @param quiet if `TRUE`, intermediary output is suppressed #' @importFrom stats coef pnbinom -#' @importFrom amap K #' @return A named vector of the genes containing outlying cells and the number #' of cells on each. setGeneric( diff --git a/R/DIscBIO-generic-KMClustDiffGenes.R b/R/DIscBIO-generic-KMClustDiffGenes.R index d6b1eda..db728e1 100644 --- a/R/DIscBIO-generic-KMClustDiffGenes.R +++ b/R/DIscBIO-generic-KMClustDiffGenes.R @@ -11,7 +11,6 @@ #' @param filename_down Name of the exported "down" file (if `export=TRUE`) #' @param filename_binom Name of the exported binomial file #' @param filename_sigdeg Name of the exported sigDEG file -#' @importFrom dplyr summarize #' @importFrom stats pbinom median #' @rdname KMClustDiffGenes #' @return A list containing two tables. @@ -159,10 +158,10 @@ setMethod( DEGsE <- c(DEGsE, as.character(rownames(Final))) Up <- subset(Final, Final[, 7] == "Up") - Up <- dplyr::select( - Up, "Regulation", "genes", "pv", "mean.all", "mean.cl", - "fc", "p.adj" + cols_to_keep <- c( + "Regulation", "genes", "pv", "mean.all", "mean.cl", "fc", "p.adj" ) + Up <- Up[, cols_to_keep] Up[, 3] <- rownames(Up) Up[, 6] <- log2(Up[, 6]) Up[, 1] <- Up[, 2] @@ -182,10 +181,10 @@ setMethod( } Down <- subset(Final, Final[, 7] == "Down") - Down <- dplyr::select( - Down, "Regulation", "genes", "pv", "mean.all", - "mean.cl", "fc", "p.adj" + cols_to_keep <- c( + "Regulation", "genes", "pv", "mean.all", "mean.cl", "fc", "p.adj" ) + Down <- Down[, cols_to_keep] Down[, 3] <- rownames(Down) Down[, 6] <- log2(Down[, 6]) Down[, 1] <- Down[, 2] diff --git a/R/DIscBIO-generic-MBClustDiffGenes.R b/R/DIscBIO-generic-MBClustDiffGenes.R index ebaf110..7931b61 100644 --- a/R/DIscBIO-generic-MBClustDiffGenes.R +++ b/R/DIscBIO-generic-MBClustDiffGenes.R @@ -155,10 +155,10 @@ setMethod( DEGsE <- c(DEGsE, as.character(rownames(Final))) Up <- subset(Final, Final[, 7] == "Up") - Up <- dplyr::select( - Up, "Regulation", "genes", "pv", "mean.all", "mean.cl", - "fc", "p.adj" + cols_to_keep <- c( + "Regulation", "genes", "pv", "mean.all", "mean.cl", "fc", "p.adj" ) + Up <- Up[, cols_to_keep] Up[, 3] <- rownames(Up) Up[, 6] <- log2(Up[, 6]) Up[, 1] <- Up[, 2] @@ -173,10 +173,10 @@ setMethod( } Down <- subset(Final, Final[, 7] == "Down") - Down <- dplyr::select( - Down, "Regulation", "genes", "pv", "mean.all", - "mean.cl", "fc", "p.adj" + cols_to_keep <- c( + "Regulation", "genes", "pv", "mean.all", "mean.cl", "fc", "p.adj" ) + Down <- Down[, cols_to_keep] Down[, 3] <- rownames(Down) Down[, 6] <- log2(Down[, 6]) Down[, 1] <- Down[, 2] diff --git a/R/DIscBIO-generic-NoiseFiltering.R b/R/DIscBIO-generic-NoiseFiltering.R index 87744e9..6ed66b2 100644 --- a/R/DIscBIO-generic-NoiseFiltering.R +++ b/R/DIscBIO-generic-NoiseFiltering.R @@ -20,7 +20,6 @@ #' excel file. Default is TRUE. #' @param quiet if `TRUE`, suppresses printed output #' @param filename Name of the exported file (if `export=TRUE`) -#' @importFrom matrixStats rowVars #' @importFrom stats quantile var fitted.values pchisq p.adjust median aggregate #' @importFrom graphics plot axis abline points lines #' @importFrom statmod glmgam.fit @@ -84,10 +83,10 @@ setMethod( # perform fit, define sample moments per gene meansG1ms <- rowMeans(nCountsG1ms) - varsG1ms <- rowVars(nCountsG1ms) + varsG1ms <- apply(nCountsG1ms, 1, var) cv2G1ms <- varsG1ms / meansG1ms ^ 2 meansERCC <- rowMeans(nCountsERCC) - varsERCC <- rowVars(nCountsERCC) + varsERCC <- apply(nCountsERCC, 1, var) cv2ERCC <- varsERCC / meansERCC ^ 2 minMeanForFit <- unname( quantile(meansERCC[which(cv2ERCC > CV)], percentile) diff --git a/R/J48DT.R b/R/J48DT.R index 21f6dcc..38e516d 100644 --- a/R/J48DT.R +++ b/R/J48DT.R @@ -8,8 +8,6 @@ #' @param plot If `FALSE`, suppresses plot output #' @importFrom RWeka J48 #' @importFrom graphics plot -#' @importFrom partykit as.party -#' @importFrom grid gpar #' @return Information about the J48 model and, by default, a plot of the #' decision tree. J48DT <- function(data, quiet = FALSE, plot = TRUE) { @@ -30,19 +28,7 @@ J48DT <- function(data, quiet = FALSE, plot = TRUE) { exp.df <- as.data.frame(t(data)) classVector <- factor(colnames(data)) j48.model <- J48(classVector ~ ., exp.df) - if (!quiet) - print(j48.model) - if (plot) { - plot( - as.party(j48.model), - gp = gpar( - cex = 0.65, - col = "black", - lty = "solid", - lwd = 1.5, - fontsize = 12 - ) - ) - } + if (!quiet) print(j48.model) + if (plot) plot(j48.model) return(j48.model) } \ No newline at end of file diff --git a/R/Jaccard.R b/R/Jaccard.R index 1680d53..4b1efb2 100644 --- a/R/Jaccard.R +++ b/R/Jaccard.R @@ -47,6 +47,7 @@ Jaccard <- function( } else if (Clustering == "MB") { target_col <- object@MBclusters$clusterid } + # TODO: replace with in-house code to eliminate boot package dependency results <- boot( data = object@fdata[, which(target_col == i)], statistic = JS, diff --git a/R/VolcanoPlot.R b/R/VolcanoPlot.R index 37d017f..101e5e3 100644 --- a/R/VolcanoPlot.R +++ b/R/VolcanoPlot.R @@ -13,7 +13,6 @@ #' samr.compute.siggenes.table #' @importFrom graphics title #' @importFrom utils write.csv -#' @importFrom calibrate textxy #' @return A volcano plot #' @export VolcanoPlot <- function(object, value = 0.05, name = NULL, fc = 0.5, FS = .4) { @@ -63,7 +62,7 @@ VolcanoPlot <- function(object, value = 0.05, name = NULL, fc = 0.5, FS = .4) { col = "blue" )) with(sigFC, - textxy( + text( abs(sigFC[, 7]), -log10(sigFC[, 8]), labs = sigFC[, 2], diff --git a/R/retrieveBiomart.R b/R/retrieveBiomart.R deleted file mode 100644 index e43aa5f..0000000 --- a/R/retrieveBiomart.R +++ /dev/null @@ -1,123 +0,0 @@ -#' @title Retrieve data from BioMart -#' @description uses functions from the biomaRt package to retrieve dataframes -#' from the BioMart Database -#' @details Since the BioMart database is not always accessible, this function -#' envelops the requests to the database in a set of tryCatch functions to -#' allow for multiple queries and easier feedback to the end user -#' @param gene_name gene signature -#' @param quiet if `TRUE`, suppresses messages -#' @param max_tries maximum number of times the function will try to reach the -#' database -#' @importFrom biomaRt useDataset useMart useEnsembl getBM -#' @return data.frame resulting from a successful call to getBM. -retrieveBiomart <- function(gene_name, quiet = FALSE, max_tries = 3) { - # Generates a Mart object - if (!quiet) - message("Retrieving mart object. Please wait.") - mart <- tryCatch({ - useDataset("hsapiens_gene_ensembl", useMart("ensembl")) - }, - error = function(err) { - message( - "The Mart object could not be retrieved.", - "Will try again using a different function.", - "Here is the original error message:\n", - err - ) - return(NULL) - }) - if (is.null(mart)) { - mart <- tryCatch({ - useDataset("hsapiens_gene_ensembl", useEnsembl("ensembl")) - }, - error = function(err) { - message(err) - stop( - "The Mart object still could not be retrieved. ", - "The server may be down. Please try again later." - ) - return(NULL) - }) - } - - # Retrieve BioMart dataframe - if (!quiet) { - message( - "Done.\n", - "Accessing the BioMart database.", - "This operation may take a few minutes. Please wait." - ) - } - G_list <- NULL - tries <- 1 - while (is.null(G_list) & tries <= max_tries) { - G_list <- tryCatch({ - if (quiet) { - suppressMessages( - getBM( - filters = "ensembl_gene_id", - attributes = c("ensembl_gene_id", "hgnc_symbol"), - values = gene_name, - mart = mart, - useCache = FALSE - ) - ) - } else { - getBM( - filters = "ensembl_gene_id", - attributes = c("ensembl_gene_id", "hgnc_symbol"), - values = gene_name, - mart = mart, - useCache = FALSE - ) - } - }, - error = function(err) { - if (!quiet) { - message( - "The BioMart database could not be reached. ", - "Retrying (", - tries, - "/", - max_tries, - "). ", - "Here is the original error message:\n", - err - ) - } - tries <- tries + 1 - return(NULL) - }, - warning = function(warn) { - if (!quiet) { - message( - "The BioMart database could not be reached. ", - "Retrying (", - tries, - "/", - max_tries, - "). ", - "Here is the original warning:\n", - warn - ) - } - tries <- tries + 1 - return(NULL) - }) - } - - # Giving up or returning output - if (is.null(G_list) & tries > 3) { - stop( - "The BioMart database could not be reached after", - max_tries, - "tries. ", - "Please try again later." - ) - } else { - if (!quiet) - message("Done.") - } - - return(G_list) - } \ No newline at end of file diff --git a/R/samr-adapted.R b/R/samr-adapted.R index ceb1fc8..98864d8 100644 --- a/R/samr-adapted.R +++ b/R/samr-adapted.R @@ -816,15 +816,15 @@ foldchange.twoclass = function(x, y, logged2) { return(fc) } #' @title Foldchange of twoclass unpaired sequencing data -#' @importFrom matrixStats rowMedians #' @param x x #' @param y y #' @param depth depth foldchange.seq.twoclass.unpaired <- function(x, y, depth) { x.norm <- scale(x, center = F, scale = depth) + 1e-08 - fc <- rowMedians(x.norm[, y == 2])/rowMedians(x.norm[, y == - 1]) + fc <- apply(x.norm[, y == 2], 1, median) / + apply(x.norm[, y == + 1], 1, median) return(fc) } foldchange.seq.twoclass.paired <- function(x, y, depth) { @@ -836,7 +836,7 @@ foldchange.seq.twoclass.paired <- function(x, y, depth) { } x.norm <- scale(x, center = F, scale = depth) + 1e-08 d <- x.norm[, o2, drop = F]/x.norm[, o1, drop = F] - fc <- rowMedians(d, na.rm = T) + fc <- lapply(d, 1, function(x) median(x, na.rm = T)) return(fc) } permute <- function(elem) { @@ -1150,8 +1150,8 @@ foldchange.paired = function(x, y, logged2) { foldchange.seq.twoclass.unpaired <- function(x, y, depth) { x.norm <- scale(x, center = F, scale = depth) + 1e-08 - fc <- rowMedians(x.norm[, y == 2])/rowMedians(x.norm[, y == - 1]) + fc <- apply(x.norm[, y == 2], 1, median) / + apply(x.norm[, y == 1, 1, median]) return(fc) } integer.base.b <- function(x, b = 2) { diff --git a/man/retrieveBiomart.Rd b/man/retrieveBiomart.Rd deleted file mode 100644 index 71cd9da..0000000 --- a/man/retrieveBiomart.Rd +++ /dev/null @@ -1,28 +0,0 @@ -% Generated by roxygen2: do not edit by hand -% Please edit documentation in R/retrieveBiomart.R -\name{retrieveBiomart} -\alias{retrieveBiomart} -\title{Retrieve data from BioMart} -\usage{ -retrieveBiomart(gene_name, quiet = FALSE, max_tries = 3) -} -\arguments{ -\item{gene_name}{gene signature} - -\item{quiet}{if `TRUE`, suppresses messages} - -\item{max_tries}{maximum number of times the function will try to reach the -database} -} -\value{ -data.frame resulting from a successful call to getBM. -} -\description{ -uses functions from the biomaRt package to retrieve dataframes - from the BioMart Database -} -\details{ -Since the BioMart database is not always accessible, this function - envelops the requests to the database in a set of tryCatch functions to - allow for multiple queries and easier feedback to the end user -} From 1ee95c9a4d817c392fc886740d7ff981f5a61189 Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Wed, 29 Jul 2020 16:11:13 +0200 Subject: [PATCH 62/99] Aggregated internal functions --- DESCRIPTION | 1 + NAMESPACE | 1 - R/DIscBIO-generic-Clustexp.R | 219 --------------------- R/DIscBIO-generic-KMClustDiffGenes.R | 6 - R/DIscBIO-generic-KMclustheatmap.R | 10 - R/DIscBIO-generic-MBClustDiffGenes.R | 6 - R/DIscBIO-generic-MBclustheatmap.R | 78 +------- R/DIscBIO-generic-NoiseFiltering.R | 18 -- R/DIscBIO-generic-Normalizedata.R | 29 --- R/DIscBIO-generic-comptSNE.R | 6 - R/DIscBIO-generic-comptsneMB.R | 7 - R/DIscBIO-generic-plotSilhouette.R | 6 - R/DIscBIO-generic-plotsilhouetteMB.R | 6 - R/J48DTeval.R | 45 ----- R/RpartEVAL.R | 46 ----- R/VolcanoPlot.R | 18 -- R/internal-functions.R | 272 +++++++++++++++++++++++++++ 17 files changed, 275 insertions(+), 499 deletions(-) create mode 100644 R/internal-functions.R diff --git a/DESCRIPTION b/DESCRIPTION index db8e912..fbd1f7b 100644 --- a/DESCRIPTION +++ b/DESCRIPTION @@ -101,6 +101,7 @@ Collate: 'VolcanoPlot.R' 'customConverters.R' 'datasets.R' + 'internal-functions.R' 'prepExampleDataset.R' 'reformatSiggenes.R' 'replaceDecimals.R' diff --git a/NAMESPACE b/NAMESPACE index b0da0b0..5c00ab0 100644 --- a/NAMESPACE +++ b/NAMESPACE @@ -66,7 +66,6 @@ importFrom(fpc,calinhara) importFrom(fpc,cluster.stats) importFrom(fpc,clusterboot) importFrom(fpc,dudahart2) -importFrom(fpc,kmeansCBI) importFrom(ggplot2,aes) importFrom(ggplot2,aes_string) importFrom(ggplot2,element_blank) diff --git a/R/DIscBIO-generic-Clustexp.R b/R/DIscBIO-generic-Clustexp.R index b4227e2..1b90ace 100644 --- a/R/DIscBIO-generic-Clustexp.R +++ b/R/DIscBIO-generic-Clustexp.R @@ -27,7 +27,6 @@ #' @param quiet if `TRUE`, intermediate output is suppressed #' @importFrom stats as.dist cor kmeans #' @importFrom cluster clusGap maxSE -#' @importFrom fpc clusterboot cluster.stats calinhara dudahart2 #' @importFrom graphics pairs #' @importFrom methods is #' @return The DISCBIO-class object input with the cpart slot filled. @@ -109,224 +108,6 @@ setMethod( cln = cln, rseed = rseed ) - dist.gen <- - function(x, method = "euclidean", ...) - if (method %in% c("spearman", "pearson", "kendall")) - as.dist(1 - cor(t(x), method = method, ...)) - else - dist(x, method = method, ...) - dist.gen.pairs <- - function(x, y, ...) - dist.gen(t(cbind(x, y)), ...) - clustfun <- - function(x, - clustnr = 20, - bootnr = 50, - metric = "pearson", - do.gap = TRUE, - SE.method = "Tibs2001SEmax", - SE.factor = .25, - B.gap = 50, - cln = 0, - rseed = rseed, - quiet = FALSE) { - if (clustnr < 2) - stop("Choose clustnr > 1") - di <- - dist.gen(t(x), method = metric) - if (do.gap | cln > 0) { - gpr <- NULL - if (do.gap) { - set.seed(rseed) - gpr <- - clusGap( - as.matrix(di), - FUNcluster = kmeans, - K.max = clustnr, - B = B.gap, - verbose = !quiet - ) - if (cln == 0) - cln <- - maxSE(gpr$Tab[, 3], - gpr$Tab[, 4], - method = SE.method, - SE.factor) - } - if (cln <= 1) { - clb <- list( - result = list( - partition = rep(1, dim(x)[2]) - ), - bootmean = 1 - ) - names(clb$result$partition) <- names(x) - return(list( - x = x, - clb = clb, - gpr = gpr, - di = di - )) - } - - Kmeansruns <- - function (data, - krange = 2:10, - criterion = "ch", - iter.max = 100, - runs = 100, - scaledata = FALSE, - alpha = 0.001, - critout = FALSE, - plot = FALSE, - method = "euclidean", - ...) { - data <- as.matrix(data) - if (criterion == "asw") - sdata <- - dist(data) - if (scaledata) - data <- - scale(data) - cluster1 <- - 1 %in% krange - crit <- - numeric(max(krange)) - km <- list() - for (k in krange) { - if (k > 1) { - minSS <- Inf - kmopt <- - NULL - for (i in 1:runs) { - opar <- options(show.error.messages = FALSE) - on.exit(options(opar)) - repeat { - kmm <- try(kmeans(data, k)) - if (!is(kmm, "try-error")) break - } - opar <- options(show.error.messages = TRUE) - on.exit(options(opar)) - swss <- - sum(kmm$withinss) - if (swss < minSS) { - kmopt <- kmm - minSS <- - swss - } - if (plot) { - opar <- par(ask = TRUE) - on.exit(par(opar)) - pairs(data, - col = kmm$cluster, - main = swss) - } - } - km[[k]] <- - kmopt - crit[k] <- - switch( - criterion, - asw = cluster.stats( - sdata, - km[[k]]$cluster - )$avg.silwidth, - ch = calinhara(data, - km[[k]]$cluster) - ) - if (critout) { - message(k, " clusters ", crit[k], "\n") - } - } - } - if (cluster1) - cluster1 <- - dudahart2( - data, km[[2]]$cluster, alpha = alpha - )$cluster1 - k.best <- - which.max(crit) - if (cluster1) - k.best <- - 1 - km[[k.best]]$crit <- - crit - km[[k.best]]$bestk <- - k.best - out <- - km[[k.best]] - out - } - - - KmeansCBI <- - function (data, - krange, - k = NULL, - scaling = FALSE, - runs = 1, - criterion = "ch", - method = "euclidean", - ...) { - if (!is.null(k)) - krange <- - k - if (!identical(scaling, FALSE)) - sdata <- - scale(data, - center = TRUE, - scale = scaling) - else - sdata <- - data - c1 <- - Kmeansruns( - sdata, - krange, - runs = runs, - criterion = criterion, - method = method, - ... - ) - partition <- - c1$cluster - cl <- list() - nc <- krange - for (i in 1:nc) - cl[[i]] <- - partition == i - out <- - list( - result = c1, - nc = nc, - clusterlist = cl, - partition = partition, - clustermethod = "kmeans" - ) - out - } - - clb <- clusterboot( - di, - B = bootnr, - distances = FALSE, - bootmethod = "boot", - clustermethod = KmeansCBI, - krange = cln, - scaling = FALSE, - multipleboot = FALSE, - bscompare = TRUE, - seed = rseed, - count = !quiet - ) - return(list( - x = x, - clb = clb, - gpr = gpr, - di = di - )) - } - } y <- clustfun( object@fdata, clustnr, diff --git a/R/DIscBIO-generic-KMClustDiffGenes.R b/R/DIscBIO-generic-KMClustDiffGenes.R index db728e1..a0bcad4 100644 --- a/R/DIscBIO-generic-KMClustDiffGenes.R +++ b/R/DIscBIO-generic-KMClustDiffGenes.R @@ -55,12 +55,6 @@ setMethod( x <- object@ndata y <- object@expdata[, names(object@ndata)] part <- object@kmeans$kpart - binompval <- function(p, N, n) { - pval <- pbinom(n, round(N, 0), p, lower.tail = TRUE) - filter <- !is.na(pval) & pval > 0.5 - pval[filter] <- 1 - pval[filter] - return(pval) - } for (i in 1:max(part)) { if (sum(part == i) == 0) next diff --git a/R/DIscBIO-generic-KMclustheatmap.R b/R/DIscBIO-generic-KMclustheatmap.R index f47e8c8..350c1b3 100644 --- a/R/DIscBIO-generic-KMclustheatmap.R +++ b/R/DIscBIO-generic-KMclustheatmap.R @@ -45,16 +45,6 @@ setMethod( tmp <- cbind(tmp, cent) } names(tmp) <- paste("cl", na, sep = ".") - - dist.gen <- - function(x, method = "euclidean", ...) - if (method %in% c("spearman", "pearson", "kendall")) - as.dist(1 - cor(t(x), method = method, ...)) - else - dist(x, method = method, ...) - dist.gen.pairs <- - function(x, y, ...) - dist.gen(t(cbind(x, y)), ...) if (max(part) > 1) cclmo <- hclust(dist.gen(as.matrix( diff --git a/R/DIscBIO-generic-MBClustDiffGenes.R b/R/DIscBIO-generic-MBClustDiffGenes.R index 7931b61..2ecfc7e 100644 --- a/R/DIscBIO-generic-MBClustDiffGenes.R +++ b/R/DIscBIO-generic-MBClustDiffGenes.R @@ -52,12 +52,6 @@ setMethod( x <- object@ndata y <- object@expdata[, names(object@ndata)] part <- object@MBclusters$clusterid - binompval <- function(p, N, n) { - pval <- pbinom(n, round(N, 0), p, lower.tail = TRUE) - filter <- !is.na(pval) & pval > 0.5 - pval[filter] <- 1 - pval[filter] - return(pval) - } for (i in 1:max(part)) { if (sum(part == i) == 0) next diff --git a/R/DIscBIO-generic-MBclustheatmap.R b/R/DIscBIO-generic-MBclustheatmap.R index 2994907..c4b056d 100644 --- a/R/DIscBIO-generic-MBclustheatmap.R +++ b/R/DIscBIO-generic-MBclustheatmap.R @@ -12,7 +12,6 @@ #' @param rseed Random integer to fix random results. #' @importFrom stats hclust as.dist cor kmeans #' @importFrom cluster clusGap maxSE -#' @importFrom fpc clusterboot kmeansCBI #' @return Unless otherwise specified, a heatmap and a vector of the underlying #' cluster order. setGeneric( @@ -30,80 +29,7 @@ setMethod( definition = function(object, hmethod, plot, quiet, rseed) { x <- object@fdata object@clusterpar$metric <- "pearson" - dist.gen <- - function(x, method = "euclidean", ...) - if (method %in% c("spearman", "pearson", "kendall")) - as.dist(1 - cor(t(x), method = method, ...)) - else - dist(x, method = method, ...) - dist.gen.pairs <- function(x, y, ...) - dist.gen(t(cbind(x, y)), ...) - clustfun <- - function(x, - clustnr = 20, - bootnr = 50, - metric = "pearson", - do.gap = TRUE, - SE.method = "Tibs2001SEmax", - SE.factor = .25, - B.gap = 50, - cln = 0, - rseed = rseed, - quiet = FALSE) { - if (clustnr < 2) - stop("Choose clustnr > 1") - di <- dist.gen(t(x), method = metric) - if (do.gap | cln > 0) { - gpr <- NULL - if (do.gap) { - set.seed(rseed) - gpr <- clusGap( - as.matrix(di), - FUNcluster = kmeans, - K.max = clustnr, - B = B.gap, - verbose = !quiet - ) - if (cln == 0) - cln <- maxSE( - gpr$Tab[, 3], gpr$Tab[, 4], method = SE.method, SE.factor - ) - } - if (cln <= 1) { - clb <- list(result = list(partition = rep(1, dim(x)[2])), - bootmean = 1) - names(clb$result$partition) <- names(x) - return(list( - x = x, - clb = clb, - gpr = gpr, - di = di - )) - } - # FUN <- match.fun(clustermethod) - clb <- clusterboot( - di, - B = bootnr, - distances = FALSE, - bootmethod = "boot", - clustermethod = fpc::kmeansCBI, - krange = cln, - scaling = FALSE, - multipleboot = FALSE, - bscompare = TRUE, - seed = rseed, - count = !quiet - ) - return(list( - x = x, - clb = clb, - gpr = gpr, - di = di - )) - } - } - y <- - clustfun( + y <- clustfun( object@fdata, clustnr = 20, bootnr = 50, @@ -115,7 +41,7 @@ setMethod( cln = 0, rseed = rseed, quiet = quiet - ) + ) object@distances <- as.matrix(y$di) part <- object@MBclusters$clusterid na <- c() diff --git a/R/DIscBIO-generic-NoiseFiltering.R b/R/DIscBIO-generic-NoiseFiltering.R index 6ed66b2..0add7d0 100644 --- a/R/DIscBIO-generic-NoiseFiltering.R +++ b/R/DIscBIO-generic-NoiseFiltering.R @@ -243,24 +243,6 @@ setMethod( col = erccCol ) # Showing all the valied ERCCs } - add_legend <- function(...) { - opar <- par( - fig = c(0, 1, 0, 1), - oma = c(0, 0, 0, 0), - mar = c(0, 0, 0, 0), - new = TRUE - ) - on.exit(par(opar)) - plot( - 0, - 0, - type = 'n', - bty = 'n', - xaxt = 'n', - yaxt = 'n' - ) - legend(...) - } add_legend( "topleft", legend = c( diff --git a/R/DIscBIO-generic-Normalizedata.R b/R/DIscBIO-generic-Normalizedata.R index ce2760c..4357069 100644 --- a/R/DIscBIO-generic-Normalizedata.R +++ b/R/DIscBIO-generic-Normalizedata.R @@ -69,35 +69,6 @@ setMethod( cols <- apply(object@expdata, 2, sum, na.rm = TRUE) >= mintotal object@ndata <- object@expdata[, cols] if (downsample) { - downsample <- function(x, n, dsn) { - x <- round(x[, apply(x, 2, sum, na.rm = TRUE) >= n], 0) - nn <- min(apply(x, 2, sum)) - for (j in 1:dsn) { - z <- data.frame(GENEID = rownames(x)) - rownames(z) <- rownames(x) - initv <- rep(0, nrow(z)) - for (i in 1:dim(x)[2]) { - y <- - aggregate(rep(1, nn), list(sample( - rep(rownames(x), x[, i]), nn - )), sum) - na <- names(x)[i] - names(y) <- c("GENEID", na) - rownames(y) <- y$GENEID - z[, na] <- initv - k <- intersect(rownames(z), y$GENEID) - z[k, na] <- y[k, na] - z[is.na(z[, na]), na] <- 0 - } - rownames(z) <- as.vector(z$GENEID) - ds <- if (j == 1) - z[, -1] - else - ds + z[, -1] - } - ds <- ds / dsn + .1 - return(ds) - } set.seed(rseed) object@ndata <- downsample(object@expdata, n = mintotal, dsn = dsn) } else{ diff --git a/R/DIscBIO-generic-comptSNE.R b/R/DIscBIO-generic-comptSNE.R index f65fabb..71754d4 100644 --- a/R/DIscBIO-generic-comptSNE.R +++ b/R/DIscBIO-generic-comptSNE.R @@ -29,12 +29,6 @@ if (length(object@kmeans$kpart) == 0) stop("run Clustexp before comptSNE") set.seed(rseed) - dist.gen <- - function(x, method = "euclidean") - if (method %in% c("spearman", "pearson", "kendall")) - as.dist(1 - cor(t(x), method = method)) - else - dist(x, method = method) di <- dist.gen(as.matrix(object@distances)) if (quiet) { ts <- suppressMessages( diff --git a/R/DIscBIO-generic-comptsneMB.R b/R/DIscBIO-generic-comptsneMB.R index e6bca24..7c0ae18 100644 --- a/R/DIscBIO-generic-comptsneMB.R +++ b/R/DIscBIO-generic-comptsneMB.R @@ -28,13 +28,6 @@ setMethod( if (length(object@MBclusters) == 0) stop("run clustexp before comptsneMB") set.seed(rseed) - dist.gen <- function(x, method = "euclidean") { - if (method %in% c("spearman", "pearson", "kendall")) { - as.dist(1 - cor(t(x), method = method)) - } else { - dist(x, method = method) - } - } di <- dist.gen(as.matrix(t(object@fdata))) if (quiet) { ts <- suppressMessages( diff --git a/R/DIscBIO-generic-plotSilhouette.R b/R/DIscBIO-generic-plotSilhouette.R index ba9be84..8d6c1f5 100644 --- a/R/DIscBIO-generic-plotSilhouette.R +++ b/R/DIscBIO-generic-plotSilhouette.R @@ -30,12 +30,6 @@ setMethod( } col <- c("black", "blue", "green", "red", "yellow", "gray") kpart <- object@kmeans$kpart - dist.gen <- - function(x, method = "euclidean", ...) - if (method %in% c("spearman", "pearson", "kendall")) - as.dist(1 - cor(t(x), method = method, ...)) - else - dist(x, method = method, ...) distances <- dist.gen(object@distances) si <- silhouette(kpart, distances) plot(si, col = col[1:K]) diff --git a/R/DIscBIO-generic-plotsilhouetteMB.R b/R/DIscBIO-generic-plotsilhouetteMB.R index 84893ba..814aae6 100644 --- a/R/DIscBIO-generic-plotsilhouetteMB.R +++ b/R/DIscBIO-generic-plotsilhouetteMB.R @@ -26,12 +26,6 @@ setMethod( stop("only a single cluster: no silhouette plot") col = c("black", "blue", "green", "red", "yellow", "gray") kpart <- object@MBclusters$clusterid - dist.gen <- - function(x, method = "euclidean", ...) - if (method %in% c("spearman", "pearson", "kendall")) - as.dist(1 - cor(t(x), method = method, ...)) - else - dist(x, method = method, ...) distances <- dist.gen(t(object@fdata)) si <- silhouette(kpart, distances) plot(si, col = col[1:K]) diff --git a/R/J48DTeval.R b/R/J48DTeval.R index 8e3758f..bdd33c8 100644 --- a/R/J48DTeval.R +++ b/R/J48DTeval.R @@ -46,25 +46,7 @@ J48DTeval <- function( } else{ stop("Unknown classification algorithm") } - #Evaluate model on test set - eval.pred <- function(pred.class, true.class, class1, - performance) { - for (index in 1:length(pred.class)) { - pred <- pred.class[index] - true <- true.class[index] - if (pred == true && true == class1) { - performance["TP"] <- performance["TP"] + 1 - } else if (pred != true && true == class1) { - performance["FN"] <- performance["FN"] + 1 - } else if (pred != true && true != class1) { - performance["FP"] <- performance["FP"] + 1 - } else if (pred == true && true != class1) { - performance["TN"] <- performance["TN"] + 1 - } - } - return(performance) - } performance <- eval.pred( pred.class, test.class, class1, performance ) @@ -92,33 +74,6 @@ J48DTeval <- function( ) colnames(j48.confusion.matrix) <- c(First, Second) if (!quiet) print(j48.confusion.matrix) - - SN <- function(con.mat) { - TP <- con.mat[1, 1] - FN <- con.mat[2, 1] - return(TP / (TP + FN)) - } - SP <- function(con.mat) { - TN <- con.mat[2, 2] - FP <- con.mat[1, 2] - return(TN / (TN + FP)) - } - ACC <- function(con.mat) { - TP <- con.mat[1, 1] - FN <- con.mat[2, 1] - TN <- con.mat[2, 2] - FP <- con.mat[1, 2] - return((TP + TN) / (TP + FN + TN + FP)) - } - MCC <- function(con.mat) { - TP <- con.mat[1, 1] - FN <- con.mat[2, 1] - TN <- con.mat[2, 2] - FP <- con.mat[1, 2] - denom <- sqrt((TP + FP) * (TP + FN) * (TN + FP) * (TN + FN)) - denom <- ifelse(denom == 0, NA, denom) - return((TP * TN - FP * FN) / denom) - } j48.sn <- SN(j48.confusion.matrix) j48.sp <- SP(j48.confusion.matrix) j48.acc <- ACC(j48.confusion.matrix) diff --git a/R/RpartEVAL.R b/R/RpartEVAL.R index 6f9da1e..e32be89 100644 --- a/R/RpartEVAL.R +++ b/R/RpartEVAL.R @@ -45,24 +45,6 @@ RpartEVAL <- function(data, num.folds = 10, First = "CL1", Second = "CL2", stop("Unknown classification algorithm") } #Evaluate model on test set - - eval.pred <- function(pred.class, true.class, class1, - performance) { - for (index in 1:length(pred.class)) { - pred <- pred.class[index] - true <- true.class[index] - if (pred == true && true == class1) { - performance["TP"] <- performance["TP"] + 1 - } else if (pred != true && true == class1) { - performance["FN"] <- performance["FN"] + 1 - } else if (pred != true && true != class1) { - performance["FP"] <- performance["FP"] + 1 - } else if (pred == true && true != class1) { - performance["TN"] <- performance["TN"] + 1 - } - } - return(performance) - } performance <- eval.pred( pred.class, test.class, class1, performance ) @@ -92,34 +74,6 @@ RpartEVAL <- function(data, num.folds = 10, First = "CL1", Second = "CL2", ) colnames(Rpart.confusion.matrix) <- c(First, Second) if (!quiet) print(Rpart.confusion.matrix) - - SN <- function(con.mat) { - TP <- con.mat[1, 1] - FN <- con.mat[2, 1] - return(TP / (TP + FN)) - } - SP <- function(con.mat) { - TN <- con.mat[2, 2] - FP <- con.mat[1, 2] - return(TN / (TN + FP)) - } - ACC <- function(con.mat) { - TP <- con.mat[1, 1] - FN <- con.mat[2, 1] - TN <- con.mat[2, 2] - FP <- con.mat[1, 2] - return((TP + TN) / (TP + FN + TN + FP)) - } - MCC <- function(con.mat) { - TP <- con.mat[1, 1] - FN <- con.mat[2, 1] - TN <- con.mat[2, 2] - FP <- con.mat[1, 2] - denom <- sqrt((TP + FP) * (TP + FN) * (TN + FP) * (TN + FN)) - denom <- ifelse(denom == 0, NA, denom) - return((TP * TN - FP * FN) / denom) - } - Rpart.sn <- SN(Rpart.confusion.matrix) Rpart.sp <- SP(Rpart.confusion.matrix) Rpart.acc <- ACC(Rpart.confusion.matrix) diff --git a/R/VolcanoPlot.R b/R/VolcanoPlot.R index 101e5e3..5d19f89 100644 --- a/R/VolcanoPlot.R +++ b/R/VolcanoPlot.R @@ -70,24 +70,6 @@ VolcanoPlot <- function(object, value = 0.05, name = NULL, fc = 0.5, FS = .4) { col = "blue" ) ) - add_legend <- function(...) { - opar <- par( - fig = c(0, 1, 0, 1), - oma = c(0, 0, 0, 0), - mar = c(0, 0, 0, 0), - new = TRUE - ) - on.exit(par(opar)) - plot( - 0, - 0, - type = 'n', - bty = 'n', - xaxt = 'n', - yaxt = 'n' - ) - legend(...) - } add_legend( "topleft", legend = c( diff --git a/R/internal-functions.R b/R/internal-functions.R new file mode 100644 index 0000000..5534575 --- /dev/null +++ b/R/internal-functions.R @@ -0,0 +1,272 @@ +#' @importFrom fpc clusterboot cluster.stats calinhara dudahart2 +clustfun <- function( + x, + clustnr = 20, + bootnr = 50, + metric = "pearson", + do.gap = TRUE, + SE.method = "Tibs2001SEmax", + SE.factor = .25, + B.gap = 50, + cln = 0, + rseed = rseed, + quiet = FALSE +) { + if (clustnr < 2) stop("Choose clustnr > 1") + di <- dist.gen(t(x), method = metric) + if (do.gap | cln > 0) { + gpr <- NULL + if (do.gap) { + set.seed(rseed) + gpr <- clusGap( + as.matrix(di), + FUNcluster = kmeans, + K.max = clustnr, + B = B.gap, + verbose = !quiet + ) + if (cln == 0) { + cln <- maxSE( + gpr$Tab[, 3], + gpr$Tab[, 4], + method = SE.method, + SE.factor + ) + } + } + if (cln <= 1) { + clb <- list( + result = list(partition = rep(1, dim(x)[2])), + bootmean = 1 + ) + names(clb$result$partition) <- names(x) + return(list(x = x, clb = clb, gpr = gpr, di = di)) + } + clb <- clusterboot( + di, + B = bootnr, + distances = FALSE, + bootmethod = "boot", + clustermethod = KmeansCBI, + krange = cln, + scaling = FALSE, + multipleboot = FALSE, + bscompare = TRUE, + seed = rseed, + count = !quiet + ) + return(list(x = x, clb = clb,gpr = gpr, di = di)) + } +} + +Kmeansruns <- function ( + data, + krange = 2: 10, + criterion = "ch", + iter.max = 100, + runs = 100, + scaledata = FALSE, + alpha = 0.001, + critout = FALSE, + plot = FALSE, + method = "euclidean", + ... +) { + data <- as.matrix(data) + if (criterion == "asw") sdata <- dist(data) + if (scaledata) data <- scale(data) + cluster1 <- 1 %in% krange + crit <- numeric(max(krange)) + km <- list() + for (k in krange) { + if (k > 1) { + minSS <- Inf + kmopt <- NULL + for (i in 1:runs) { + opar <- options(show.error.messages = FALSE) + on.exit(options(opar)) + repeat { + kmm <- try(kmeans(data, k)) + if (!is(kmm, "try-error")) break + } + opar <- options(show.error.messages = TRUE) + on.exit(options(opar)) + swss <- sum(kmm$withinss) + if (swss < minSS) { + kmopt <- kmm + minSS <- swss + } + if (plot) { + opar <- par(ask = TRUE) + on.exit(par(opar)) + pairs(data, col = kmm$cluster, main = swss) + } + } + km[[k]] <- kmopt + crit[k] <- switch( + criterion, + asw = cluster.stats(sdata, km[[k]]$cluster)$avg.silwidth, + ch = calinhara(data, km[[k]]$cluster) + ) + if (critout) { + message(k, " clusters ", crit[k], "\n") + } + } + } + if (cluster1) { + cluster1 <- dudahart2( data, km[[2]]$cluster, alpha = alpha)$cluster1 + } + k.best <- which.max(crit) + if (cluster1) k.best <- 1 + km[[k.best]]$crit <- crit + km[[k.best]]$bestk <- k.best + out <- km[[k.best]] + return(out) +} + +KmeansCBI <- function (data, + krange, + k = NULL, + scaling = FALSE, + runs = 1, + criterion = "ch", + method = "euclidean", + ... +) { + if (!is.null(k)) krange <- k + if (!identical(scaling, FALSE)) { + sdata <- scale(data, center = TRUE, scale = scaling) + } else { + sdata <- data + } + c1 <- Kmeansruns( + sdata, + krange, + runs = runs, + criterion = criterion, + method = method, + ... + ) + partition <- c1$cluster + cl <- list() + nc <- krange + for (i in 1:nc) cl[[i]] <- partition == i + out <- list( + result = c1, + nc = nc, + clusterlist = cl, + partition = partition, + clustermethod = "kmeans" + ) + return(out) +} + +dist.gen <- function(x, method = "euclidean") { + if (method %in% c("spearman", "pearson", "kendall")) { + as.dist(1 - cor(t(x), method = method)) + } else { + dist(x, method = method) + } +} + +binompval <- function(p, N, n) { + pval <- pbinom(n, round(N, 0), p, lower.tail = TRUE) + filter <- !is.na(pval) & pval > 0.5 + pval[filter] <- 1 - pval[filter] + return(pval) +} + +add_legend <- function(...) { + opar <- par( + fig = c(0, 1, 0, 1), + oma = c(0, 0, 0, 0), + mar = c(0, 0, 0, 0), + new = TRUE + ) + on.exit(par(opar)) + plot( + x = 0, + y = 0, + type = 'n', + bty = 'n', + xaxt = 'n', + yaxt = 'n' + ) + legend(...) +} + +downsample <- function(x, n, dsn) { + x <- round(x[, apply(x, 2, sum, na.rm = TRUE) >= n], 0) + nn <- min(apply(x, 2, sum)) + for (j in 1:dsn) { + z <- data.frame(GENEID = rownames(x)) + rownames(z) <- rownames(x) + initv <- rep(0, nrow(z)) + for (i in 1:dim(x)[2]) { + y <- aggregate( + rep(1, nn), list(sample( + rep(rownames(x), x[, i]), nn + )), + sum + ) + na <- names(x)[i] + names(y) <- c("GENEID", na) + rownames(y) <- y$GENEID + z[, na] <- initv + k <- intersect(rownames(z), y$GENEID) + z[k, na] <- y[k, na] + z[is.na(z[, na]), na] <- 0 + } + rownames(z) <- as.vector(z$GENEID) + ds <- if (j == 1) z[, -1] else ds + z[, -1] + } + ds <- ds / dsn + .1 + return(ds) +} + +eval.pred <- function(pred.class, true.class, class1, performance) { + for (index in 1:length(pred.class)) { + pred <- pred.class[index] + true <- true.class[index] + if (pred == true && true == class1) { + performance["TP"] <- performance["TP"] + 1 + } else if (pred != true && true == class1) { + performance["FN"] <- performance["FN"] + 1 + } else if (pred != true && true != class1) { + performance["FP"] <- performance["FP"] + 1 + } else if (pred == true && true != class1) { + performance["TN"] <- performance["TN"] + 1 + } + } + return(performance) +} + +SN <- function(con.mat) { + TP <- con.mat[1, 1] + FN <- con.mat[2, 1] + return(TP / (TP + FN)) +} + +SP <- function(con.mat) { + TN <- con.mat[2, 2] + FP <- con.mat[1, 2] + return(TN / (TN + FP)) +} + +ACC <- function(con.mat) { + TP <- con.mat[1, 1] + FN <- con.mat[2, 1] + TN <- con.mat[2, 2] + FP <- con.mat[1, 2] + return((TP + TN) / (TP + FN + TN + FP)) +} + +MCC <- function(con.mat) { + TP <- con.mat[1, 1] + FN <- con.mat[2, 1] + TN <- con.mat[2, 2] + FP <- con.mat[1, 2] + denom <- sqrt((TP + FP) * (TP + FN) * (TN + FP) * (TN + FN)) + denom <- ifelse(denom == 0, NA, denom) + return((TP * TN - FP * FN) / denom) +} \ No newline at end of file From cf251439e26fc6d489fca0fb2ad3a377127af58b Mon Sep 17 00:00:00 2001 From: Waldir Leoncio Date: Wed, 29 Jul 2020 17:30:10 +0200 Subject: [PATCH 63/99] Removed vignette --- vignettes/.gitignore | 2 - vignettes/Clustering.png | Bin 144502 -> 0 bytes vignettes/DIscBIO-vignette.Rmd | 356 ------------------------------- vignettes/Data-Preprocessing.png | Bin 86473 -> 0 bytes vignettes/DiscBIO.png | Bin 134077 -> 0 bytes vignettes/KM1.png | Bin 103106 -> 0 bytes vignettes/KM2.png | Bin 104089 -> 0 bytes vignettes/KM3.png | Bin 96695 -> 0 bytes vignettes/MB1.png | Bin 103552 -> 0 bytes vignettes/MB2.png | Bin 104138 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rmarkdown::html_vignette -vignette: > - %\VignetteIndexEntry{DIscBIO tutorial} - %\VignetteEngine{knitr::rmarkdown} - %\VignetteEncoding{UTF-8} ---- -```{r options, echo=FALSE} -knitr::opts_chunk$set(fig.width=7, fig.height=7, eval=FALSE) -``` - -The pipeline consists of four successive steps: data pre-processing, cellular clustering and pseudo-temporal ordering, determining differential expressed genes and identifying biomarkers. - -This vignette is a reduced version of the interactive notebook available [here](https://mybinder.org/v2/gh/SystemsBiologist/PSCAN/discbio-pub?filepath=DIscBIO.ipynb) - -## Loading dataset - -The dataset should be formatted in a data frame where columns refer to samples and rows refer to genes. - -```{r} -library(DIscBIO) -FileName <- data("valuesG1msRed") -DataSet <- get(FileName) -``` - -## 1. Data Pre-processing - -Prior to applying data analysis methods, it is standard to pre-process the raw read counts resulted from the sequencing. The preprocessing approach depends on the existence or absence of ERCC spike-ins. In both cases, it includes normalization of read counts and gene filtering. - -#### Normalization of read counts -To account for RNA composition and sequencing depth among samples (single-cells), the normalization method “median of ratios” is used. This method takes the ratio of the gene instantaneous median to the total counts for all genes in that cell (column median). The gene instantaneous median is the product of multiplying the median of the total counts across all cells (row median) with the read of the target gene in each cell. This normalization method makes it possible to compare the normalized counts for each gene equally between samples. - -#### Gene filtering -The key idea in filtering genes is to appoint the genes that manifest abundant variation across samples. Filtering genes is a critical step due to its dramatic impact on the downstream analysis. In case the raw data includes ERCC spike-ins, genes will be filtered based on variability in comparison to a noise level estimated from the ERCC spike-ins according to an algorithm developed by Brennecke et al (Brennecke et al., 2013). This algorithm utilizes the dependence of technical noise on the average read count and fits a model to the ERCC spike-ins. Further gene filtering can be implemented based on gene expression. -In case the raw data does not include ERCC spike-ins, genes will be only filtered based on minimum expression in certain number of cells. - -```{r} -sc <- DISCBIO(DataSet) -``` - -### 1.1. Filtering and normalizing the raw data that includes ERCCs - -Filtering the raw data that includes ERCCs can be done by applying the “NoiseFiltering” function, which includes several parameters: - - - object: the outcome of running the DISCBIO() function. - - percentile: A numeric value of the percentile. It is used to validate the ERCC spik-ins. Default is 0.8. - - CV: A numeric value of the coefficient of variation. It is used to validate the ERCC spik-ins. Default is 0.5. - - geneCol: Color of the genes that did not pass the filtration. - - FgeneCol: Color of the genes that passt the filtration. - - erccCol: Color of the ERCC spik-ins. - - Val: A logical vector that allows plotting only the validated ERCC spike-ins. Default is TRUE. If Val=FALSE will - plot all the ERCC spike-ins. - - plot: A logical vector that allows plotting the technical noise. Default is TRUE. - - export: A logical vector that allows writing the final gene list in excel file. Default is TRUE. - - quiet: if `TRUE`, suppresses printed output - -To normalize the raw sequencing reads the function Normalizedata() should be used, this function takes 8 parameters. -- In case the user would like just to normalize the reads without any further gene filtering the parameters minexpr and minnumber should be set to 0. -- In case the user would like just to normalize the reads and run gene filtering based on gene expression the parameters minexpr and minnumber should have values. This function will discard cells with less than mintotal transcripts. Genes that are not expressed at minexpr transcripts in at least minnumber cells are discarded. - -The function Normalizedata() normalizes the count reads using the normalization method “median of ratios” - -To Finalize the preprocessing the function FinalPreprocessing() should be implemented by setting the parameter "GeneFlitering" to NoiseF ( whether the dditional gene filtering step based on gene expression was implemented on not). - - -```{r} -sc <- NoiseFiltering(sc,percentile=0.9, CV=0.2) - -#### Normalizing the reads without any further gene filtering -sc <- Normalizedata(sc, mintotal=1000, minexpr=0, minnumber=0, maxexpr=Inf, downsample=FALSE, dsn=1, rseed=17000) - -sc <- FinalPreprocessing(sc,GeneFlitering="NoiseF") ### The GeneFiltering can be either "NoiseF" or"ExpF" -``` - -### 1.2. Filtering and normalizing the raw data that does not include ERCCs - -To normalize and filter the raw data that does not include ERCCs can be done by applying the function Normalizedata() and giving the parameters minexpr and minnumber some values. This function will discard cells with less than mintotal transcripts. Genes that are not expressed at minexpr transcripts in at least minnumber cells are discarded. Furthermore, it will normalize the count reads using the normalization method “median of ratios”. - -To Finalize the preprocessing the function FinalPreprocessing() should be implemented by setting the parameter "GeneFlitering" to ExpF. - -## 2. Cellular Clustering and Pseudo Time ordering - -Cellular clustering is performed according to the gene expression profiles to detect cellular sub-population with unique properties. After clustering, pseudo-temporal ordering is generated to indicate the cellular differentiation degree. - -## 2.1. K-means Clustering - -Rare cell type Identification algorithm (RaceID) was used to cluster the pre-processed data using k-means on a similarity distance matrix, which was based on Pearson correlation and the similarity matrix was computed as “1 – Pearson correlation”. The approach of the proposed clustering, i.e., applying k-means on a similarity distance matrix using the Euclidean metric, improves cluster separation. RaceID estimates the number of clusters by finding the minimal clusters' number at the saturation level of gap statistics, which standardizes the within-cluster dispersion. - -The Clustexp() functions has several parameters: -- object: the outcome of running the DISCBIO() function. -- clustnr Maximum number of clusters for the derivation of the cluster number by the saturation of mean within-cluster dispersion. Default is 20. -- bootnr A numeric value of booststrapping runs for \code{clusterboot}. Default is 50. -- metric Is the method to transform the input data to a distance object. -- Metric has to be one of the following: ["spearman","pearson","kendall","euclidean","maximum","manhattan","canberra","binary","minkowski"]. -- do.gap A logical vector that allows generating the number of clusters based on the gap statistics. Default is TRUE. -- SE.method The SE.method determines the first local maximum of the gap statistics. -- The SE.method has to be one of the following:["firstSEmax","Tibs2001SEmax","globalSEmax","firstmax","globalmax"]. Default is "Tibs2001SEmax" -- SE.factor A numeric value of the fraction of the standard deviation by which the local maximum is required to differ from the neighboring points it is compared to. Default is 0.25. -- B.gap Number of bootstrap runs for the calculation of the gap statistics. Default is 50 -- cln Number of clusters to be used. Default is \code{NULL} and the cluster number is inferred by the saturation criterion. -- rseed Integer number. Random seed to enforce reproducible clustering results. Default is 17000. -- quiet if `TRUE`, intermediate output is suppressed - -```{r} -sc <- Clustexp(sc,cln=2,quiet=TRUE) #### K-means clustering to get three clusters -plotGap(sc) ### Plotting gap statisticssc <- Clustexp(sc, clustnr=20,bootnr=50,metric="pearson",do.gap=TRUE,SE.method="Tibs2001SEmax",SE.factor=.25,B.gap=50,cln=2,rseed=17000) -``` - -#### 2.1.1. Defining the Cells in the clusters generated by k-means clustering - - -```{r} -sc <- comptSNE(sc,rseed=15555,quiet=TRUE) -sc@cpart -``` - -#### 2.1.2- Evaluating the stability and consistency of the clusters - -PSCAN enables the robustness assessment of the detected clusters in terms of stability and consistency using Jaccard’s similarity statistics and silhouette coefficients. Jaccard’s similarity index provides a comparison of members among clusters to evaluate the stability of the clusters with a range from 0% to 100%. The higher the percentage, the more stable the cluster is. Silhouette coefficients estimate how close each sample in one cluster is to samples in the neighboring clusters, reflecting the consistency of each cluster with a range of [-1, 1]. The higher the cluster mean coefficient, the more consistent the cluster is. - - -```{r} -# Silhouette of k-means clusters -# you might have to run "par(mar=c(6,2,4,2))" first so plot looks better -plotSilhouette(sc,K=2) # K is the number of clusters -``` - -```{r} -Jaccard(sc,Clustering="K-means", K=2, plot=TRUE) # Jaccard of k-means clusters -``` - -#### 2.1.3. Cluster plotting using tSNE maps - -Here you visualize the K-means clusters using t-distributed stochastic neighbor embedding (tSNE), which is a non-linear dimensionality reduction method that places neighbor cells close to each other. - - -```{r} -############ Plotting K-means clusters -plottSNE(sc) -plotKmeansLabelstSNE(sc) # To plot the the ID of the cells in each cluster -plotSymbolstSNE(sc,types=sub("(\\_\\d+)$","", names(sc@ndata))) # To plot the the ID of the cells in each cluster -``` - -### Defining outlier cells based on K-means Clustering - -Outlier identification is implemented using a background model based on distribution of transcript counts within a cluster. Outlier cells are detected if the probability for that cell c that a minimum number of genes Gmin of observing total counts TGmin is less than a specific threshold Pthr, as given by the red dotted line. Outlier cells in K-means clusters can be detected by running the FindOutliersKM() function. - -In case the user decided to remove outlier cells, the user should set RemovingOutliers to TRUE and then start from the beginning (Data Pre-processing). - - -```{r} -outlg <- round(length(sc@fdata[,1])/200) # The cell will be considered as an outlier if it has a minimum of 0.5% of the number of filtered genes as outlier genes. -Outliers <- FindOutliersKM(sc, K=2, outminc=5,outlg=outlg,probthr=.5*1e-3,thr=2**-(1:40),outdistquant=.75, plot=TRUE, quiet=FALSE) -``` - -#### 2.1.4. Cellular pseudo-time ordering based on k-means clusters - -```{r} -sc <- KmeanOrder(sc,quiet=FALSE, export=FALSE) -plotOrderKMtsne(sc) -``` - -#### 2.1.5 Plotting the K-means clusters in heatmap - -The similarities between single-cells were computed by Euclidean distances of Pearson transcriptome correlation matrix. Based on these similarities, a heatmap portrayal of cell-to-cell distances was plotted using Euclidean as the distance measure and the single linkage as the clustering method, cluster centers were ordered by hierarchic clustering. - - -```{r} -KMclustheatmap(sc,hmethod="single", plot=TRUE) -``` - -#### 2.1.6 Plotting the gene expression of a particular gene in a tSNE map - - -```{r} -plotExptSNE(sc, "ENSG00000101346") -``` - -## 3. Determining differentially expressed genes (DEGs) For K-means Clustering - -Differentially expressed genes between individual clusters are identified using the significance analysis of sequencing data (SAMseq), which is a new function in significance analysis of microarrays (Li and Tibshirani 2011) in the samr package v2.0 (Tibshirani et all., 2015). SAMseq is a non-parametric statistical function dependent on Wilcoxon rank statistic that equalizes the sizes of the library by a resampling method accounting for the various sequencing depths. The analysis is implemented over the pure raw dataset that has the unnormalized expression read counts after excluding the ERCCs. Furthermore, DEGs in each cluster comparing to all the remaining clusters are determined using binomial differential expression, which is based on binomial counting statistics. - -## 3.1 Identifying DEGs using SAMseq - -The user can define DEGs between all clusters generated by either K-means or model based clustering by applying the “DEGanalysis” function. Another alternative is to define DEGs between particular clusters generated by K-means clustering by applying the “DEGanalysis2clust” function. The outcome of these two functions is a list of two components: -- The first component is a data frame showing the Ensembl gage name and the symbole of the detected DEGs -- The second component is table showing the comparisons, Target cluster, Number of genes and the File name. This component will be used for the downstream analysis. - -### 3.1.1 Determining DEGs between two particular clusters - -```{r degKM} -####### differential expression analysis between cluster 1 and cluster 3 of the Model-Based clustering using FDR of 0.05 -cdiff <- DEGanalysis2clust(sc,Clustering="K-means",K=2,fdr=0.05,name="Name",First="CL1",Second="CL2",export=FALSE,quiet=TRUE) -``` - -```{r} -#### To show the result table -head(cdiff[[1]]) # The first component -head(cdiff[[2]]) # The second component -``` - -### 3.1.2 Determining DEGs between all clusters - - -**¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤ Running this cell will overwrite the previous one ¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤¤** - - -```{r} -cdiff <- DEGanalysis(sc,Clustering="K-means",K=2,fdr=0.05,name="all_clusters",export=FALSE,quiet=TRUE) ####### differential expression analysis between all clusters -``` - -```{r} -#### To show the result table -head(cdiff[[1]]) # The first component -head(cdiff[[2]]) # The second component -``` - -#### 3.2 Identifying DEGs using binomial differential expression - -The function KMClustDiffGenes identifies differentially regulated genes for each cluster of the K-means clustering in -comparison to the ensemble of all cells. It returns a list with a data.frame element for each cluster that contains the mean expression across all cells not in the cluster (mean.ncl) and in the cluster (mean.cl), the fold-change in the cluster versus all remaining cells (fc), and the p-value for differential expression between all cells in a cluster and all remaining cells. The p-value is computed based on the overlap of negative binomials fitted to the count distributions within the two groups akin to DESeq. - - -```{r} -cdiffBinomial <- KMClustDiffGenes(sc,K=2,fdr=.01,,quiet=TRUE) ########## Binomial differential expression analysis -#### To show the result table -head(cdiffBinomial[[1]]) # The first component -head(cdiffBinomial[[2]]) # The second component -``` - -### Plotting the DEGs - -Volcano plots are used to readily show the DEGs by plotting significance versus fold-change on the y and x axes, respectively. - -```{r} -name <- cdiffBinomial[[2]][2, 4] # Selecting the DEGs table -U <- read.csv(file=paste0(name), head=TRUE, sep=",") -Vplot <- VolcanoPlot(U, value=0.001, name=name, FS=.5, fc=1) -``` - -In case the output of this function is the error "Error in plot.window(...): need finite 'ylim' values", `adj` should be set to TRUE. - -In case the user would like to get the names of the significant DEGs, then FS should be set to a value of 0.3 or higher. - -## 4. Identifying biomarkers (decision trees and networking analysis) - -There are several methods to identify biomarkers, among them are decision trees and hub detection through networking analysis. The outcome of STRING analysis is stored in tab separated values (TSV) files. These TSV files served as an input to check both the connectivity degree and the betweenness centrality, which reflects the communication flow in the defined PPI networks - -Decision trees are one of the most efficient classification techniques in biomarkers discovery. Here we use it to predict the sub-population of a target cell based on transcriptomic data. Two types of decision trees can be performed: classification and regression trees (CART) and J48. The decision tree analysis is implemented over a training dataset, which consisted of the DEGs obtained by either SAMseq or the binomial differential expression. The performance of the generated trees can be evaluated for error estimation by ten-fold cross validation assessment using the "J48DTeval" and "RpartEVAL" functions. The decision tree analysis requires the dataset to be class vectored by applying the “ClassVectoringDT” function. - -```{r} -#sigDEG <- cdiff[[1]] # DEGs gene list from SANseq -sigDEG <- cdiffBinomial[[1]] # DEGs gene list from Binomial analysis -First="CL1" -Second="CL2" -DATAforDT <- ClassVectoringDT(sc,Clustering="K-means",K=2,First=First,Second=Second,sigDEG) -``` - -### 4.1. J48 Decision Tree - - -```{r} -j48dt <- J48DT(DATAforDT) -summary(j48dt) -``` - -#### 4.1.1. Evaluating the performance of the J48 Decision Tree - - -```{r} -j48dt <- J48DTeval(DATAforDT,num.folds=10,First=First,Second=Second) -``` - -### 4.2. RPART Decision Tree - - -```{r} -rpartDT <- RpartDT(DATAforDT) -``` - -#### 4.2.1. Evaluating the performance of the RPART Decision Tree - - -```{r} -rpartEVAL <- RpartEVAL(DATAforDT,num.folds=10,First=First,Second=Second) -``` - -### 4.3. Networking Analysis - -To define protein-protein interactions (PPI) over a list of genes, STRING-api is used. The outcome of STRING analysis was stored in tab separated values (TSV) files. These TSV files served as an input to check both the connectivity degree and the betweenness centrality, which reflects the communication flow in the defined PPI networks. - -### 4.3.1 All DEGs - - -```{r} -DEGs="All DEGs" -FileName=paste0(DEGs) - -data <- cdiffBinomial[[1]] [,2] # DEGs gene list from Binomial analysis (taking only the firat 100 genes) - -ppi <- PPI(data,FileName) - -networking <- NetAnalysis(ppi) -networking ##### In case the Examine response components=200 and an error "linkmat[i, ]" appeared, that means there are no PPI. -``` - -```{r} -data=networking[1:25,1] # plotting the network of the top 25 highly connected genes -network <- Networking(data,FileName,plot_width=25, plot_height=10) -``` - -### 4.3.2 Particular set of DEGs - - -```{r} -############ Selecting the DEGs' table ############## -DEGs=cdiffBinomial[[2]][1,4] # Up-regulated genes in cluster 1 (from the Binomial analysis) -FileName=paste0(DEGs) - -data <- read.csv(file=paste0(DEGs),head=TRUE,sep=",") -data <- data[,3] - -ppi <- PPI(data,FileName) - -networking <- NetAnalysis(ppi) -networking ##### In case the Examine response components=200 and an error "linkmat[i, ]" appeared, that means there are no PPI. - -network <- Networking(data,FileName) -``` - -### 4.4 Gene Enrichment Analysis - -```{r} -dbs <- listEnrichrDbs() -head(dbs) -``` - -```{r} -############ Selecting the DEGs' table ############## -DEGs=cdiffBinomial[[2]][1,4] # Up-regulated genes in cluster 1 (from the Binomial analysis) - -data <- read.csv(file=paste0(DEGs),head=TRUE,sep=",") -data <- as.character(data[,3]) - -dbs <- c("KEGG_2013","GO_Molecular_Function_2015", "GO_Biological_Process_2015") -enriched <- enrichr(data, dbs) -KEGG_2013 <- enriched[[1]][,c(1,2,3,9)] -GO_Molecular_Function_2015 <- enriched[[2]][,c(1,2,3,9)] -GO_Biological_Process_2015 <- enriched[[3]][,c(1,2,3,9)] - -GEA <- rbind(KEGG_2013,GO_Molecular_Function_2015,GO_Biological_Process_2015) -GEA -``` diff --git a/vignettes/Data-Preprocessing.png 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