This repository contains all codes used to analyse the data reported in "Reprogramming by Drug-like Molecules Leads to Regeneration of Cochlear Hair Cells in Adult Mice"

https://www.pnas.org/doi/10.1073/pnas.2215253120

Authors

Yi-Zhou Quan1,2†, Wei Wei1,2,4†, Volkan Ergin1,2†, Arun Prabhu Rameshbabu1,2, Mingqian Huang1,2, Chunjie Tian1,2, Srinivas Vinod Saladi3,5, Artur A Indzhykulian1,2, and Zheng-Yi Chen1,2*

Affiliations

1 Department of Otolaryngology-Head and Neck Surgery, Graduate Program in Speech and Hearing Bioscience and Technology and Program in Neuroscience, Harvard Medical School, Boston, MA 02115.

2 Eaton-Peabody Laboratory, Massachusetts Eye and Ear Infirmary, 243 Charles St., Boston, MA 02114

3 Broad Institute of MIT and Harvard, Cambridge, Massachusetts 02142, USA.

4 Department of Otolaryngology-Head and Necks, Shengjing Hospital of China Medical University, Shenyang, 110004, China

5 Department of Otolaryngology Head and Neck Surgery, Massachusetts Eye and Ear Infirmary, Boston, Massachusetts.

†These authors contributed equally

*Corresponding author: Zheng-Yi_Chen@meei.harvard.edu

ABSTRACT

Strategies to overcome irreversible cochlear hair cell (HC) damage and loss in mammals are of vital importance to hearing recovery in patients with permanent hearing loss. In mature mammalian cochlea, co-activation of Myc and Notch1 reprograms the supporting cells (SC) and promotes HC regeneration. Understanding of the underlying mechanisms may aid the development of a clinically-relevant approach to achieve HC regeneration in wildtype mature cochlea. By single-cell RNAseq, we show that MYC/NICD “rejuvenates” the adult mouse cochlea by activating the pathways including Wnt and cAMP, whose blockade suppresses HC regeneration. We screened and identified a combination (the cocktail) of drug-like molecules composing of small molecules and siRNAs to activate the pathways of Myc, Notch1, Wnt and cAMP. We show that the cocktail effectively replaces Myc and Notch1 transgenes to reprogram fully mature wildtype supporting cells for HC regeneration in vitro. Finally, we demonstrated the cocktail is capable of reprogramming adult cochlea for HC regeneration in a mouse model with hair cell loss in vivo. Our study identified a strategy by a clinically-relevant approach to reprogram mature inner ear for HC regeneration, laying the foundation for hearing restoration by HC regeneration.