Effects of genetic correction on the differentiation of hair cell-like cells from iPSCs with MYO15A mutation

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.

Pluripotent stem cell-derived cochlear cells: a challenge in constant progress

“…However, these differentiated cells failed to exhibit stereociliary hair bundles at their apical surface [51,52], indicating that these factors were not sufficient to drive the full program of hair cell maturation. However, electrophysiologically functional hair cell-like cells were derived from OEPs when co-cultured with chicken utricle stromal cells complemented or not with RA and EGF treatment [52][53][54], demonstrating that, as for mES cells, chicken utricle stromal cells provide factors required for human hair cell development. Adding a step of PPR induction prior to the otic induction step also gave rise to hair cell-like cells [55].…”

“…3 Comparision of published protocols of human ES and iPS cell differentiation into hair cell-like cells. BMP bone morphogenetic protein, EB embryoid bodies, EGF epidermal growth factor, FGF fibroblast growth factor, hESCs human embryonic stem cells, hPSCs human induced pluripotent sten cells, IGF-1 insulin-like growth factor 2, LIF leukemia inhibitory factor, NNE non neural ectoderm, OEPD otic epibranchial placode domain, PPR pre placodal region, RA retinoic acid, TCF-β transforming growth factor-β promising step for treating human hereditary deafness [53,54]. Indeed, CRISPR/CAS9 system was successfully used to correct deafness-associated mutations in hiPSCs.…”

“…The stem cell-derived hair cell-like cells that present mutation in MYO7 or MYO15 gene harboured defects in stereocilia bundles. The correction of gene mutation using CRISPR/ CAS9 technology restored the morphology of hair bundles and the functionality of the derived hair cells [53,54].…”

Induced Pluripotent Stem Cells Meet Genome Editing

“…For example, the genetic correction of mutations in Niemann-Pick Type C patient-specific iPSCs to rescue metabolic defects in cholesterol metabolism and autophagy, which are responsible for the pathology, represents just one demonstration of how this approach has been successfully implemented (Maetzel et al, 2014). Furthermore, genome editing in hPSCs has been used to establish models for Rett-syndrome disrupting MECP2 function in hPSCs (Li et al, 2013), to generate HIV-resistant variants alleles of the CCR5 gene into iPSCs (Ye et al, 2014), to repair MYO15A in iPSCs derived from patients affected by deafness (Chen et al, 2016) and to derive isogeneic cell pairs of COL7A1-corrected iPSCs derived from patients with dystrophic epidermolysis bullosa (Sebastiano et al, 2014). …”

Recent Advancements in Gene and Stem Cell‐Based Treatment Modalities: Potential Implications in Noise‐Induced Hearing Loss

“…iPSCs generated from deaf patients with a congenital MYO7A and MYO15A mutation were genetically corrected using CRISPR/Cas9. These modified iPSCs, CP‐iPSCs, demonstrated pluripotency and differentiated into hair cell‐like cells that exhibited restored stereociliary organization as well as electrophysiological function (Chen et al, ; Tang et al, ).…”