Dual expression of Atoh1 and Ikzf2 promotes transformation of adult cochlear supporting cells into outer hair cells

  1. Suhong Sun
  2. Shuting Li
  3. Zhengnan Luo
  4. Minhui Ren
  5. Shunji He
  6. Guangqin Wang
  7. Zhiyong Liu  Is a corresponding author
  1. Chinese Academy of Sciences, China

Abstract

Mammalian cochlear outer hair cells (OHCs) are essential for hearing. Severe hearing impairment follows OHC degeneration. Previous attempts at regenerating new OHCs from cochlear supporting cells (SCs) have been unsuccessful, notably lacking expression of the key OHC motor protein, Prestin. Thus, regeneration of Prestin+ OHCs represents a barrier to restore auditory function in vivo. Here, we reported the successful in vivo conversion of adult mouse cochlear SCs into Prestin+ OHC-like cells through the concurrent induction of two key transcriptional factors known to be necessary for OHC development: Atoh1 and Ikzf2. Single cell RNA sequencing revealed the upregulation of 729 OHC genes and downregulation of 331 SC genes in OHC-like cells. The resulting differentiation status of these OHC-like cells was much more advanced than previously achieved. This study thus established an efficient approach to induce the regeneration of Prestin+ OHCs, paving the way for in vivo cochlear repair via SC transdifferentiation.

Data availability

Sequencing data have been deposited in GEO under accession codes: GSE161156.

The following previously published data sets were used

Article and author information

Author details

  1. Suhong Sun

    Institute of Neuroscience, Chinese Academy of Sciences, Shanghai, China
    Competing interests
    Suhong Sun, We filed an auditory hair cell regeneration patent based on the key findings of this manuscript..
  2. Shuting Li

    Institute of Neuroscience, Chinese Academy of Sciences, Shanghai, China
    Competing interests
    Shuting Li, We filed an auditory hair cell regeneration patent based on the key findings of this manuscript..
  3. Zhengnan Luo

    Institute of Neuroscience, Chinese Academy of Sciences, Shanghai, China
    Competing interests
    Zhengnan Luo, We filed an auditory hair cell regeneration patent based on the key findings of this manuscript..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-8204-6277
  4. Minhui Ren

    Institute of Neuroscience, Chinese Academy of Sciences, Shanghai, China
    Competing interests
    Minhui Ren, We filed an auditory hair cell regeneration patent based on the key findings of this manuscript..
  5. Shunji He

    Institute of Neuroscience, Chinese Academy of Sciences, Shanghai, China
    Competing interests
    Shunji He, We filed an auditory hair cell regeneration patent based on the key findings of this manuscript..
  6. Guangqin Wang

    Institute of Neuroscience, State Key Laboratory of Neuroscience,CAS Center for Excellence in Brain Science and Intelligence Technology, Chinese Academy of Sciences, Shanghai, China
    Competing interests
    Guangqin Wang, We filed an auditory hair cell regeneration patent based on the key findings of this manuscript..
  7. Zhiyong Liu

    Institute of Neuroscience, Chinese Academy of Sciences, Shanghai, China
    For correspondence
    Zhiyongliu@ion.ac.cn
    Competing interests
    Zhiyong Liu, We filed an auditory hair cell regeneration patent based on the key findings of this manuscript..
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-9675-1233

Funding

National Natural Science Foundation of China (81771012)

  • Zhiyong Liu

Ministry of Science and Technology of the People's Republic of China (2017YFA0103901)

  • Zhiyong Liu

Chinese Academy of Sciences (XDB32060100)

  • Zhiyong Liu

Shanghai Municipal Bureau of Quality and Technical Supervision (2018SHZDZX05)

  • Zhiyong Liu

Shanghai Jiao Tong University (SSMU-ZLCX20180601)

  • Zhiyong Liu

Boehringer Ingelheim (DE811138149)

  • Zhiyong Liu

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Ethics

Animal experimentation: All mice were bred and raised in SPF level animal rooms and animal procedures were performed according to guidelines (NA-032-2019) of the IACUC of Institute of Neuroscience (ION), Chinese Academy of Sciences.

Reviewing Editor

  1. Doris K Wu, NIDCD, NIH, United States

Publication history

  1. Received: January 14, 2021
  2. Preprint posted: January 21, 2021 (view preprint)
  3. Accepted: September 2, 2021
  4. Accepted Manuscript published: September 3, 2021 (version 1)
  5. Version of Record published: September 14, 2021 (version 2)

Copyright

© 2021, Sun et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 1,508
    Page views
  • 329
    Downloads
  • 11
    Citations

Article citation count generated by polling the highest count across the following sources: Crossref, PubMed Central, Scopus.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Suhong Sun
  2. Shuting Li
  3. Zhengnan Luo
  4. Minhui Ren
  5. Shunji He
  6. Guangqin Wang
  7. Zhiyong Liu
(2021)
Dual expression of Atoh1 and Ikzf2 promotes transformation of adult cochlear supporting cells into outer hair cells
eLife 10:e66547.
https://doi.org/10.7554/eLife.66547

Further reading

    1. Cell Biology
    2. Developmental Biology
    Juan Lu et al.
    Research Article Updated

    Phosphatidylinositol 4-phosphate (PI4P) and phosphatidylinositol 4,5-biphosphate (PIP2) are key phosphoinositides that determine the identity of the plasma membrane (PM) and regulate numerous key biological events there. To date, mechanisms regulating the homeostasis and dynamic turnover of PM PI4P and PIP2 in response to various physiological conditions and stresses remain to be fully elucidated. Here, we report that hypoxia in Drosophila induces acute and reversible depletion of PM PI4P and PIP2 that severely disrupts the electrostatic PM targeting of multiple polybasic polarity proteins. Genetically encoded ATP sensors confirmed that hypoxia induces acute and reversible reduction of cellular ATP levels which showed a strong real-time correlation with the levels of PM PI4P and PIP2 in cultured cells. By combining genetic manipulations with quantitative imaging assays we showed that PI4KIIIα, as well as Rbo/EFR3 and TTC7 that are essential for targeting PI4KIIIα to PM, are required for maintaining the homeostasis and dynamic turnover of PM PI4P and PIP2 under normoxia and hypoxia. Our results revealed that in cells challenged by energetic stresses triggered by hypoxia, ATP inhibition and possibly ischemia, dramatic turnover of PM PI4P and PIP2 could have profound impact on many cellular processes including electrostatic PM targeting of numerous polybasic proteins.

    1. Developmental Biology
    2. Neuroscience
    Mariah L Hoye et al.
    Research Article

    Mutations in the RNA helicase, DDX3X, are a leading cause of Intellectual Disability and present as DDX3X syndrome, a neurodevelopmental disorder associated with cortical malformations and autism. Yet, the cellular and molecular mechanisms by which DDX3X controls cortical development are largely unknown. Here, using a mouse model of Ddx3x loss-of-function we demonstrate that DDX3X directs translational and cell cycle control of neural progenitors, which underlies precise corticogenesis. First, we show brain development is sensitive to Ddx3x dosage; complete Ddx3x loss from neural progenitors causes microcephaly in females, whereas hemizygous males and heterozygous females show reduced neurogenesis without marked microcephaly. In addition, Ddx3x loss is sexually dimorphic, as its paralog, Ddx3y, compensates for Ddx3x in the developing male neocortex. Using live imaging of progenitors, we show that DDX3X promotes neuronal generation by regulating both cell cycle duration and neurogenic divisions. Finally, we use ribosome profiling in vivo to discover the repertoire of translated transcripts in neural progenitors, including those which are DDX3X-dependent and essential for neurogenesis. Our study reveals invaluable new insights into the etiology of DDX3X syndrome, implicating dysregulated progenitor cell cycle dynamics and translation as pathogenic mechanisms.