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

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.

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

  • 2,735
    views
  • 522
    downloads
  • 56
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

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

Share this article

https://doi.org/10.7554/eLife.66547

Further reading

    1. Developmental Biology
    Cora Demler, John C Lawlor ... Natasza A Kurpios
    Research Article

    Correct intestinal morphogenesis depends on the early embryonic process of gut rotation, an evolutionarily conserved program in which a straight gut tube elongates and forms into its first loops. However, the gut tube requires guidance to loop in a reproducible manner. The dorsal mesentery (DM) connects the gut tube to the body and directs the lengthening gut into stereotypical loops via left-right (LR) asymmetric cellular and extracellular behavior. The LR asymmetry of the DM also governs blood and lymphatic vessel formation for the digestive tract, which is essential for prenatal organ development and postnatal vital functions including nutrient absorption. Although the genetic LR asymmetry of the DM has been extensively studied, a divider between the left and right DM has yet to be identified. Setting up LR asymmetry for the entire body requires a Lefty1+ midline barrier to separate the two sides of the embryo, without it, embryos have lethal or congenital LR patterning defects. Individual organs including the brain, heart, and gut also have LR asymmetry, and while the consequences of left and right signals mixing are severe or even lethal, organ-specific mechanisms for separating these signals remain poorly understood. Here, we uncover a midline structure composed of a transient double basement membrane, which separates the left and right halves of the embryonic chick DM during the establishment of intestinal and vascular asymmetries. Unlike other basement membranes of the DM, the midline is resistant to disruption by intercalation of Netrin4 (Ntn4). We propose that this atypical midline forms the boundary between left and right sides and functions as a barrier necessary to establish and protect organ asymmetry.

    1. Developmental Biology
    Valeria Sulzyk, Ludmila Curci ... Patricia S Cuasnicu
    Research Article

    Numerous reports showed that the epididymis plays key roles in the acquisition of sperm fertilizing ability but its contribution to embryo development remains less understood. Female mice mated with males with simultaneous mutations in Crisp1 and Crisp3 genes exhibited normal in vivo fertilization but impaired embryo development. In this work, we found that this phenotype was not due to delayed fertilization, and it was observed in eggs fertilized by epididymal sperm either in vivo or in vitro. Of note, eggs fertilized in vitro by mutant sperm displayed impaired meiotic resumption unrelated to Ca2+ oscillations defects during egg activation, supporting potential sperm DNA defects. Interestingly, cauda but not caput epididymal mutant sperm exhibited increased DNA fragmentation, revealing that DNA integrity defects appear during epididymal transit. Moreover, exposing control sperm to mutant epididymal fluid or to Ca2+-supplemented control fluid significantly increased DNA fragmentation. This, together with the higher intracellular Ca2+ levels detected in mutant sperm, supports a dysregulation in Ca2+ homeostasis within the epididymis and sperm as the main factor responsible for embryo development failure. These findings highlight the contribution of the epididymis beyond fertilization and identify CRISP1 and CRISP3 as novel factors essential for sperm DNA integrity and early embryo development.