Abstract
Deafness or hearing loss is a major issue in human health. Inner ear hair cells are the main sensory receptors responsible for hearing. Defects in hair cells are one of the major causes of deafness. A combination of induced pluripotent stem cell (iPSC) technology with genome-editing technology may provide an attractive cell-based strategy to regenerate hair cells and treat hereditary deafness in humans. Here, we report the generation of iPSCs from members of a Chinese family carrying MYO15A c.4642G>A and c.8374G>A mutations and the induction of hair cell-like cells from those iPSCs. The compound heterozygous MYO15A mutations resulted in abnormal morphology and dysfunction of the derived hair cell-like cells. We used a CRISPR/Cas9 approach to genetically correct the MYO15A mutation in the iPSCs and rescued the morphology and function of the derived hair cell-like cells. Our data demonstrate the feasibility of generating inner ear hair cells from human iPSCs and the functional rescue of gene mutation-based deafness by using genetic correction.
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Abbreviations
- iPSC:
-
induced pluripotent stem cell
- ESC:
-
embryonic stem cell
- CRISPR:
-
clustered regularly interspaced short palindromic repeat
- Cas9:
-
CRISPR-associated protein 9
- HDR:
-
homology directed repair
- CUSC:
-
chicken utricle stromal cell
- AP:
-
alkaline phosphatase
- ssODN:
-
single-strand oligonucleotide
- GFP:
-
green fluorescent protein
- FACS:
-
fluorescence-activated cell sorting
- WT:
-
wild type
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Acknowledgements
We would like to thank Professor Chris Wood in Zhejiang University for critical reading of the manuscript. This study was supported by the grants from National Basic Research Program of China (2012CB967902 and 2014CB541705), National Development Program of Important Scientific Instruments (2013YQ030595), Strategically Guiding Scientific Special Projects from The Chinese Academy of Sciences (XDA04020202-23), Opening Foundation of the State Key Laboratory of Space Medicine Fundamentals and Applications (SMFA12K02), TZ-1 Application Program (KYTZ01-0901-FB-003), and The Chinese National Science Foundation (81570932 and 81330024). Also with grateful thanks for help of Miss Yu-Qin Luo Y.Q.L during karyotype analysis.
Author contributions
JRC, ZHT, JD, XDQ, CZ, JLC, CCW, LL and JZS performed the experiments and contributed to data analysis. JZ, JZC and TSH collected the clinical data. HSS and SKY performed the electrophysiological recordings. JFW, MXG, and PC drafted the conception of the study, designed the experiments, and monitored the project progression, data analysis, and interpretation. JRC prepared the initial draft of the manuscript. JFW prepared the final version of the manuscript.
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Chen, JR., Tang, ZH., Zheng, J. et al. Effects of genetic correction on the differentiation of hair cell-like cells from iPSCs with MYO15A mutation. Cell Death Differ 23, 1347–1357 (2016). https://doi.org/10.1038/cdd.2016.16
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DOI: https://doi.org/10.1038/cdd.2016.16
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