High-resolution transcriptional dissection of in vivo Atoh1-mediated hair cell conversion in mature cochleae identifies Isl1 as a co-reprogramming factor.

• Main Authors: Tetsuji Yamashita, Fei Zheng, David Finkelstein, Zoe Kellard, Robert Carter, Celeste D Rosencrance, Ken Sugino, John Easton, Charles Gawad, Jian Zuo
• Format: Article
• Language: English
• Published: Public Library of Science (PLoS) 2018-07-01
• Series: PLoS Genetics
• Online Access: http://europepmc.org/articles/PMC6086484?pdf=render
• ISSN: 1553-7390; 1553-7404

In vivo direct conversion of differentiated cells holds promise for regenerative medicine; however, improving the conversion efficiency and producing functional target cells remain challenging. Ectopic Atoh1 expression in non-sensory supporting cells (SCs) in mouse cochleae induces their partial conversion to hair cells (HCs) at low efficiency. Here, we performed single-cell RNA sequencing of whole mouse sensory epithelia harvested at multiple time points after conditional overexpression of Atoh1. Pseudotemporal ordering revealed that converted HCs (cHCs) are present along a conversion continuum that correlates with both endogenous and exogenous Atoh1 expression. Bulk sequencing of isolated cell populations and single-cell qPCR confirmed 51 transcription factors, including Isl1, are differentially expressed among cHCs, SCs and HCs. In transgenic mice, co-overexpression of Atoh1 and Isl1 enhanced the HC conversion efficiency. Together, our study shows how high-resolution transcriptional profiling of direct cell conversion can identify co-reprogramming factors required for efficient conversion.