A universal gene correction approach for FKRP-associated dystroglycanopathies to enable autologous cell therapy

Mutations in the fukutin-related protein (FKRP) gene result in a broad spectrum of muscular dystrophy (MD) phenotypes, including the severe Walker-Warburg syndrome (WWS). Here, we develop a gene-editing approach that replaces the entire mutant open reading frame with the wild-type sequence to universally correct all FKRP mutations. We apply this approach to correct FKRP mutations in induced pluripotent stem (iPS) cells derived from patients displaying broad clinical severity. Our findings show rescue of functional α-dystroglycan (α-DG) glycosylation in gene-edited WWS iPS cell-derived myotubes. Transplantation of gene-corrected myogenic progenitors in the FKRPP448L-NSG mouse model gives rise to myofiber and satellite cell engraftment and, importantly, restoration of α-DG functional glycosylation in vivo. These findings suggest the potential feasibility of using CRISPR-Cas9 technology in combination with patient-specific iPS cells for the future development of autologous cell transplantation for FKRP-associated MDs. [Display omitted] •Defective α-DG functional glycosylation in patient-specific FKRP mutant myotubes•Universal CRISPR-Cas9-mediated gene-editing approach to correct FKRP mutations•Rescue of α-DG functional glycosylation in gene-corrected FKRP myotubes•Donor-derived myofibers show rescue of α-DG functional glycosylation in FKRP mice Dhoke et al. develop a universal CRISPR-Cas9 gene-editing strategy for the correction of FKRP mutations. Myogenic progenitors from gene-edited WWS patient-specific iPS cells show rescue of phenotype in vitro and upon transplantation into muscles of FKRP mutant mice.