High-throughput Screening on Cochlear Organoids Identifies VEGFR-MEK-TGFB1 Signaling Promoting Hair Cell Reprogramming

Overview

Journal Stem Cell Reports

Publisher Cell Press

Specialty Cell Biology

Date 2021 Sep 15

PMID 34525385

Citations 11

Authors

Qing Liu

Linqing Zhang

Min-Sheng Zhu

Guoqiang Wan

Affiliations

Soon will be listed here.

Abstract

Hair cell degeneration is a major cause of sensorineural hearing loss. Hair cells in mammalian cochlea do not spontaneously regenerate, posing a great challenge for restoration of hearing. Here, we establish a robust, high-throughput cochlear organoid platform that facilitates 3D expansion of cochlear progenitor cells and differentiation of hair cells in a temporally regulated manner. High-throughput screening of the FDA-approved drug library identified regorafenib, a VEGFR inhibitor, as a potent small molecule for hair cell differentiation. Regorafenib also promotes reprogramming and maturation of hair cells in both normal and neomycin-damaged cochlear explants. Mechanistically, inhibition of VEGFR suppresses TGFB1 expression via the MEK pathway and TGFB1 downregulation directly mediates the effect of regorafenib on hair cell reprogramming. Our study not only demonstrates the power of a cochlear organoid platform in high-throughput analyses of hair cell physiology but also highlights VEGFR-MEK-TGFB1 signaling crosstalk as a potential target for hair cell regeneration and hearing restoration.

Citing Articles

Modeling, applications and challenges of inner ear organoid.

Qi J, Zhang L, Wang X, Chen X, Li Y, Wang T Smart Med. 2024; 3(1):e20230028.

PMID: 39188517 PMC: 11235738. DOI: 10.1002/SMMD.20230028.

Metabolic Profiling of Cochlear Organoids Identifies α-Ketoglutarate and NAD as Limiting Factors for Hair Cell Reprogramming.

Liu Q, Zhang L, Chen Z, He Y, Huang Y, Qiu C Adv Sci (Weinh). 2024; 11(34):e2308032.

PMID: 38993037 PMC: 11425867. DOI: 10.1002/advs.202308032.

Organoids-the key to novel therapies for the inner ear?.

Diensthuber M, Stover T HNO. 2024; 72(Suppl 2):83-88.

PMID: 38775829 DOI: 10.1007/s00106-023-01367-x.

MEK/ERK signaling drives the transdifferentiation of supporting cells into functional hair cells by modulating the Notch pathway.

Ma J, Xia M, Guo J, Li W, Sun S, Chen B Stem Cells Transl Med. 2024; 13(7):661-677.

PMID: 38709826 PMC: 11227976. DOI: 10.1093/stcltm/szae030.

[Organoids-the key to novel therapies for the inner ear? German version].

Diensthuber M, Stover T HNO. 2023; 71(11):702-707.

PMID: 37845538 DOI: 10.1007/s00106-023-01366-y.

References

Yu Y, Feng X . TGF-β signaling in cell fate control and cancer. Curr Opin Cell Biol. 2019; 61:56-63. DOI: 10.1016/j.ceb.2019.07.007. View

Lee S, Han S, Park S . Long-Term Culture of Intestinal Organoids. Methods Mol Biol. 2018; 1817:123-135. DOI: 10.1007/978-1-4939-8600-2_13. View

Fettiplace R, Hackney C . The sensory and motor roles of auditory hair cells. Nat Rev Neurosci. 2005; 7(1):19-29. DOI: 10.1038/nrn1828. View

Doerflinger N, Macklin W, Popko B . Inducible site-specific recombination in myelinating cells. Genesis. 2002; 35(1):63-72. DOI: 10.1002/gene.10154. View

Olusanya B, Neumann K, Saunders J . The global burden of disabling hearing impairment: a call to action. Bull World Health Organ. 2014; 92(5):367-73. PMC: 4007124. DOI: 10.2471/BLT.13.128728. View

Koehler K, Nie J, Longworth-Mills E, Liu X, Lee J, Holt J . Generation of inner ear organoids containing functional hair cells from human pluripotent stem cells. Nat Biotechnol. 2017; 35(6):583-589. PMC: 5462862. DOI: 10.1038/nbt.3840. View

Kuo B, Baldwin E, Layman W, Taketo M, Zuo J . In Vivo Cochlear Hair Cell Generation and Survival by Coactivation of β-Catenin and Atoh1. J Neurosci. 2015; 35(30):10786-98. PMC: 4518053. DOI: 10.1523/JNEUROSCI.0967-15.2015. View

Wan G, Corfas G, Stone J . Inner ear supporting cells: rethinking the silent majority. Semin Cell Dev Biol. 2013; 24(5):448-59. PMC: 4005836. DOI: 10.1016/j.semcdb.2013.03.009. View

Mellado Lagarde M, Wan G, Zhang L, Gigliello A, McInnis J, Zhang Y . Spontaneous regeneration of cochlear supporting cells after neonatal ablation ensures hearing in the adult mouse. Proc Natl Acad Sci U S A. 2014; 111(47):16919-24. PMC: 4250150. DOI: 10.1073/pnas.1408064111. View

10.

Wilhelm S, Dumas J, Adnane L, Lynch M, Carter C, Schutz G . Regorafenib (BAY 73-4506): a new oral multikinase inhibitor of angiogenic, stromal and oncogenic receptor tyrosine kinases with potent preclinical antitumor activity. Int J Cancer. 2010; 129(1):245-55. DOI: 10.1002/ijc.25864. View

11.

Cross M, Dixelius J, Matsumoto T, Claesson-Welsh L . VEGF-receptor signal transduction. Trends Biochem Sci. 2003; 28(9):488-94. DOI: 10.1016/S0968-0004(03)00193-2. View

12.

Jiang L, Xu J, Jin R, Bai H, Zhang M, Yang S . Transcriptomic analysis of chicken cochleae after gentamicin damage and the involvement of four signaling pathways (Notch, FGF, Wnt and BMP) in hair cell regeneration. Hear Res. 2018; 361:66-79. DOI: 10.1016/j.heares.2018.01.004. View

13.

Coffin A, Ou H, Owens K, Santos F, Simon J, Rubel E . Chemical screening for hair cell loss and protection in the zebrafish lateral line. Zebrafish. 2010; 7(1):3-11. PMC: 2935285. DOI: 10.1089/zeb.2009.0639. View

14.

Roccio M, Edge A . Inner ear organoids: new tools to understand neurosensory cell development, degeneration and regeneration. Development. 2019; 146(17). PMC: 6765123. DOI: 10.1242/dev.177188. View

15.

Schmitz F, Konigstorfer A, Sudhof T . RIBEYE, a component of synaptic ribbons: a protein's journey through evolution provides insight into synaptic ribbon function. Neuron. 2001; 28(3):857-72. DOI: 10.1016/s0896-6273(00)00159-8. View

16.

Schaefer S, Higashi A, Loomis B, Schrepfer T, Wan G, Corfas G . From Otic Induction to Hair Cell Production: Pax2 Cell Line Illuminates Key Stages of Development in Mouse Inner Ear Organoid Model. Stem Cells Dev. 2017; 27(4):237-251. PMC: 5813733. DOI: 10.1089/scd.2017.0142. View

17.

Samarajeewa A, Jacques B, Dabdoub A . Therapeutic Potential of Wnt and Notch Signaling and Epigenetic Regulation in Mammalian Sensory Hair Cell Regeneration. Mol Ther. 2019; 27(5):904-911. PMC: 6520458. DOI: 10.1016/j.ymthe.2019.03.017. View

18.

Bas E, Anwar M, van de Water T . TGF β-1 and WNT Signaling Pathways Collaboration Associated with Cochlear Implantation Trauma-Induced Fibrosis. Anat Rec (Hoboken). 2019; 303(3):608-618. DOI: 10.1002/ar.24064. View

19.

Michel O, Hess A, Bloch W, Schmidt A, Stennert E, Addicks K . Immunohistochemical detection of vascular endothelial growth factor (VEGF) and VEGF receptors Flt-1 and KDR/Flk-1 in the cochlea of guinea pigs. Hear Res. 2001; 155(1-2):175-80. DOI: 10.1016/s0378-5955(01)00262-3. View

20.

Wagner E, Shin J . Mechanisms of Hair Cell Damage and Repair. Trends Neurosci. 2019; 42(6):414-424. PMC: 6556399. DOI: 10.1016/j.tins.2019.03.006. View