Gene-edited primary muscle stem cells rescue dysferlin-deficient muscular dystrophy

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Item Type:	Preprint
Title:	Gene-edited primary muscle stem cells rescue dysferlin-deficient muscular dystrophy
Creators Name:	Escobar Fernandez, H., Di Francescantonio, S., Marg, A., Zhogov, A., Krishna, S., Metzler, E., Petkova, M., Daumke, O., Kühn, R. and Spuler, S.
Abstract:	Dystrophy-associated fer-1-like protein (dysferlin) conducts plasma membrane repair. Mutations in the DYSF gene cause a panoply of genetic muscular dystrophies. We targeted a frequent loss-of-function, DYSF exon 44, founder frameshift mutation with mRNA-mediated delivery of SpCas9 in combination with a mutation-specific sgRNA to primary muscle stem cells from two homozygous patients. We observed a consistent >60% exon 44 re-framing, rescuing a full-length and functional dysferlin protein. A new mouse model harboring a humanized Dysf exon 44 with the founder mutation, hEx44mut, recapitulated the patients’ phenotype and an identical re-framing outcome in primary muscle stem cells. Finally, gene-edited murine primary muscle stem-cells were able to regenerate muscle and rescued dysferlin when transplanted back into hEx44mut hosts. These findings are the first to show that a CRISPR-mediated therapy can ameliorate dysferlin deficiency. We suggest that gene-edited primary muscle stem cells could exhibit utility, not only in treating dysferlin deficiency syndromes, but also perhaps other forms of muscular dystrophy.
Keywords:	Animals, Mice
Source:	bioRxiv
Publisher:	Cold Spring Harbor Laboratory Press
Article Number:	2024.02.12.579813
Date:	12 February 2024
Official Publication:	https://doi.org/10.1101/2024.02.12.579813

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