title: Genetic Correction of Induced Pluripotent Stem Cells From a Deaf Patient With MYO7A Mutation Results in Morphologic and Functional Recovery of the Derived Hair Cell-Like Cells.
abstract: The genetic correction of induced pluripotent stem cells (iPSCs) induced from somatic cells of patients with sensorineural hearing loss (caused by hereditary factors) is a promising method for its treatment. The correction of gene mutations in iPSCs could restore the normal function of cells and provide a rich source of cells for transplantation. In the present study, iPSCs were generated from a deaf patient with compound heterozygous MYO7A mutations (c.1184G>A and c.4118C>T; P-iPSCs), the asymptomatic father of the patient (MYO7A c.1184G>A mutation; CF-iPSCs), and a normal donor (MYO7A(WT/WT); C-iPSCs). One of MYO7A mutation sites (c.4118C>T) in the P-iPSCs was corrected using CRISPR/Cas9. The corrected iPSCs (CP-iPSCs) retained cell pluripotency and normal karyotypes. Hair cell-like cells induced from CP-iPSCs showed restored organization of stereocilia-like protrusions; moreover, the electrophysiological function of these cells was similar to that of cells induced from C-iPSCs and CF-iPSCs. These results might facilitate the development of iPSC-based gene therapy for genetic disorders.
  
  
  CONCLUSIONS
  Induced pluripotent stem cells (iPSCs) were generated from a deaf patient with compound heterozygous MYO7A mutations (c.1184G>A and c.4118C>T). One of the MYO7A mutation sites (c.4118C>T) in the iPSCs was corrected using CRISPR/Cas9. The genetic correction of MYO7A mutation resulted in morphologic and functional recovery of hair cell-like cells derived from iPSCs. These findings confirm the hypothesis that MYO7A plays an important role in the assembly of stereocilia into stereociliary bundles. Thus, the present study might provide further insight into the pathogenesis of sensorineural hearing loss and facilitate the development of therapeutic strategies against monogenic disease through the genetic repair of patient-specific iPSCs.
authors.affiliation: Institute of Cell and Development, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, People's Republic of China.
authors.forename: Zi-Hua
authors.initials: ZH
authors.lastname: Tang
authors.affiliation: Institute of Cell and Development, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, People's Republic of China.
authors.forename: Jia-Rong
authors.initials: JR
authors.lastname: Chen
authors.affiliation: Institute of Genetics, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, People's Republic of China.
authors.forename: Jing
authors.initials: J
authors.lastname: Zheng
authors.affiliation: Department of Otorhinolaryngology, Shanghai Jiao Tong University Affiliated Sixth People's Hospital Shanghai, People's Republic of China.
authors.forename: Hao-Song
authors.initials: HS
authors.lastname: Shi
authors.affiliation: Institute of Cell and Development, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, People's Republic of China.
authors.forename: Jie
authors.initials: J
authors.lastname: Ding
authors.affiliation: The Affiliated Women's Hospital, Zhejiang University, Hangzhou, Zhejiang, People's Republic of China.
authors.forename: Xiao-Dan
authors.initials: XD
authors.lastname: Qian
authors.affiliation: Institute of Cell and Development, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, People's Republic of China.
authors.forename: Cui
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Institute of Cell and Development, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, People's Republic of China.
authors.forename: Jian-Ling
authors.initials: JL
authors.lastname: Chen
authors.affiliation: Institute of Cell and Development, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, People's Republic of China.
authors.forename: Cui-Cui
authors.initials: CC
authors.lastname: Wang
authors.affiliation: Institute of Cell and Development, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, People's Republic of China.
authors.forename: Liang
authors.initials: L
authors.lastname: Li
authors.affiliation: Department of Otolaryngology, The Affiliated Wenling People's Hospital, Wenzhou Medical University, Wenling, Zhejiang, People's Republic of China.
authors.forename: Jun-Zhen
authors.initials: JZ
authors.lastname: Chen
authors.affiliation: Department of Otorhinolaryngology, Shanghai Jiao Tong University Affiliated Sixth People's Hospital Shanghai, People's Republic of China.
authors.forename: Shan-Kai
authors.initials: SK
authors.lastname: Yin
authors.affiliation: Division of Human Genetics, Cincinnati Children's Hospital Medical Center, Cincinnati, Ohio, USA.
authors.forename: Tao-Sheng
authors.initials: TS
authors.lastname: Huang
authors.affiliation: Departments of Cell Biology and Otolaryngology, Emory University School of Medicine, Atlanta, Georgia, USA.
authors.forename: Ping
authors.initials: P
authors.lastname: Chen
authors.affiliation: Institute of Genetics, School of Medicine, Zhejiang University, Hangzhou, Zhejiang, People's Republic of China.
authors.forename: Min-Xin
authors.initials: MX
authors.lastname: Guan
authors.affiliation: Institute of Cell and Development, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, People's Republic of China wjfu@zju.edu.cn.
authors.forename: Jin-Fu
authors.initials: JF
authors.lastname: Wang
mesh_terms: D064113:CRISPR-Cas Systems; D002454:Cell Differentiation; D002460:Cell Line; D048430:Cell Shape; D004252:DNA Mutational Analysis; D005260:Female; D005786:Gene Expression Regulation; D020022:Genetic Predisposition to Disease; D006198:Hair Cells, Auditory / Q000378:metabolism / Q000637:transplantation / Q000648:ultrastructure; D006319:Hearing Loss, Sensorineural / Q000175:diagnosis / Q000235:genetics* / Q000473:pathology / Q000601:surgery; D040941:Heredity; D006579:Heterozygote; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000378:metabolism / Q000637:transplantation / Q000648:ultrastructure; D008297:Male; D008564:Membrane Potentials; D009154:Mutation; D000081412:Myosin VIIa; D009218:Myosins / Q000235:genetics*; D010375:Pedigree; D010641:Phenotype; D020127:Recovery of Function; D052416:Targeted Gene Repair / Q000379:methods*; D014162:Transfection
chemical_list: C000631455:MYO7A protein, human; D000081412:Myosin VIIa; D009218:Myosins
keywords: Deafness; Genetic correction; Human induced pluripotent stem cells; Inner ear hair cells; MYO7A; Rescue