Somatic gene editing ameliorates skeletal and cardiac muscle failure in pig and human models of Duchenne muscular dystrophy
Top Cited Papers
- 26 January 2020
- journal article
- research article
- Published by Springer Science and Business Media LLC in Nature Medicine
- Vol. 26 (2), 207-214
- https://doi.org/10.1038/s41591-019-0738-2
Abstract
Frameshift mutations in the DMD gene, encoding dystrophin, cause Duchenne muscular dystrophy (DMD), leading to terminal muscle and heart failure in patients. Somatic gene editing by sequence-specific nucleases offers new options for restoring the DMD reading frame, resulting in expression of a shortened but largely functional dystrophin protein. Here, we validated this approach in a pig model of DMD lacking exon 52 of DMD (DMDΔ52), as well as in a corresponding patient-derived induced pluripotent stem cell model. In DMDΔ52 pigs1, intramuscular injection of adeno-associated viral vectors of serotype 9 carrying an intein-split Cas9 (ref. 2) and a pair of guide RNAs targeting sequences flanking exon 51 (AAV9-Cas9-gE51) induced expression of a shortened dystrophin (DMDΔ51–52) and improved skeletal muscle function. Moreover, systemic application of AAV9-Cas9-gE51 led to widespread dystrophin expression in muscle, including diaphragm and heart, prolonging survival and reducing arrhythmogenic vulnerability. Similarly, in induced pluripotent stem cell-derived myoblasts and cardiomyocytes of a patient lacking DMDΔ52, AAV6-Cas9-g51-mediated excision of exon 51 restored dystrophin expression and amelioreate skeletal myotube formation as well as abnormal cardiomyocyte Ca2+ handling and arrhythmogenic susceptibility. The ability of Cas9-mediated exon excision to improve DMD pathology in these translational models paves the way for new treatment approaches in patients with this devastating disease.Keywords
This publication has 55 references indexed in Scilit:
- Adenoviral Vectors Coated with PAMAM Dendrimer Conjugates Allow CAR Independent Virus Uptake and Targeting to the EGF ReceptorMolecular Pharmaceutics, 2013
- Potential of primary kidney cells for somatic cell nuclear transfer mediated transgenesis in pigBMC Biotechnology, 2012
- Cardiac AAV9-S100A1 Gene Therapy Rescues Post-Ischemic Heart Failure in a Preclinical Large Animal ModelScience Translational Medicine, 2011
- Systemic Administration of PRO051 in Duchenne's Muscular DystrophyNew England Journal of Medicine, 2011
- In Vivo Canine Muscle Function AssayJournal of Visualized Experiments, 2011
- Physiological Characterization of Muscle Strength With Variable Levels of Dystrophin Restoration in mdx Mice Following Local Antisense TherapyMolecular Therapy, 2011
- Theoretic applicability of antisense-mediated exon skipping for Duchenne muscular dystrophy mutationsHuman Mutation, 2009
- Analysis of AAV Serotypes 1–9 Mediated Gene Expression and Tropism in Mice After Systemic InjectionMolecular Therapy, 2008
- Entries in the Leiden Duchenne muscular dystrophy mutation database: An overview of mutation types and paradoxical cases that confirm the reading‐frame ruleMuscle & Nerve, 2006
- Comprehensive Detection of Genomic Duplications and Deletions in the DMD Gene, by Use of Multiplex Amplifiable Probe HybridizationAmerican Journal of Human Genetics, 2002