High-resolution transcriptional dissection of in vivo Atoh1-mediated hair cell conversion in mature cochleae identifies Isl1 as a co-reprogramming factor.
Tetsuji YamashitaFei ZhengDavid FinkelsteinZoe KellardRobert CarterCeleste D RosencranceKen SuginoJohn EastonCharles GawadJian Min ZuoPublished in: PLoS genetics (2018)
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.
Keyphrases
- single cell
- rna seq
- induced apoptosis
- high resolution
- cell cycle arrest
- transcription factor
- high throughput
- gene expression
- endoplasmic reticulum stress
- cell proliferation
- cell therapy
- oxidative stress
- stem cells
- machine learning
- mass spectrometry
- bone marrow
- mesenchymal stem cells
- long non coding rna
- binding protein
- genome wide
- heat stress