A universal gene correction approach for FKRP-associated dystroglycanopathies to enable autologous cell therapy
Neha R. Dhoke,
Hyunkee Kim,
Sridhar Selvaraj,
Karim Azzag,
Haowen Zhou,
Nelio A.J. Oliveira,
Sudheer Tungtur,
Carolina Ortiz-Cordero,
James Kiley,
Qi Long Lu,
Anne G. Bang,
Rita C.R. Perlingeiro
Affiliations
Neha R. Dhoke
Lillehei Heart Institute, Department of Medicine, University of Minnesota, Minneapolis, MN, USA
Hyunkee Kim
Lillehei Heart Institute, Department of Medicine, University of Minnesota, Minneapolis, MN, USA
Sridhar Selvaraj
Lillehei Heart Institute, Department of Medicine, University of Minnesota, Minneapolis, MN, USA
Karim Azzag
Lillehei Heart Institute, Department of Medicine, University of Minnesota, Minneapolis, MN, USA
Haowen Zhou
Conrad Prebys Center for Chemical Genomics, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA, USA
Nelio A.J. Oliveira
Lillehei Heart Institute, Department of Medicine, University of Minnesota, Minneapolis, MN, USA
Sudheer Tungtur
Lillehei Heart Institute, Department of Medicine, University of Minnesota, Minneapolis, MN, USA
Carolina Ortiz-Cordero
Lillehei Heart Institute, Department of Medicine, University of Minnesota, Minneapolis, MN, USA
James Kiley
Lillehei Heart Institute, Department of Medicine, University of Minnesota, Minneapolis, MN, USA
Qi Long Lu
McColl-Lockwood Laboratory for Muscular Dystrophy Research, Cannon Research Center, Carolinas Medical Center, Atrium Health, Charlotte, NC, USA
Anne G. Bang
Conrad Prebys Center for Chemical Genomics, Sanford Burnham Prebys Medical Discovery Institute, La Jolla, CA, USA
Rita C.R. Perlingeiro
Lillehei Heart Institute, Department of Medicine, University of Minnesota, Minneapolis, MN, USA; Stem Cell Institute, University of Minnesota, Minneapolis, MN, USA; Corresponding author
Summary: 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.
