1. Autologous Cell Therapy Approach for Duchenne Muscular Dystrophy using PiggyBac Transposons and Mesoangioblasts.
- Author
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Iyer PS, Mavoungou LO, Ronzoni F, Zemla J, Schmid-Siegert E, Antonini S, Neff LA, Dorchies OM, Jaconi M, Lekka M, Messina G, and Mermod N
- Subjects
- Animals, Cell Line, Disease Models, Animal, Dystrophin genetics, Fluorescent Antibody Technique, Gene Dosage, Gene Expression, Gene Order, Genes, Reporter, Male, Mice, Mice, Inbred mdx, Mice, SCID, Muscular Dystrophy, Duchenne pathology, Muscular Dystrophy, Duchenne physiopathology, Muscular Dystrophy, Duchenne therapy, Phenotype, Transgenes, Transplantation, Autologous, Cell- and Tissue-Based Therapy methods, DNA Transposable Elements, Gene Transfer Techniques, Genetic Vectors genetics, Muscular Dystrophy, Duchenne genetics, Myoblasts metabolism, Myoblasts transplantation
- Abstract
Duchenne muscular dystrophy (DMD) is a lethal muscle-wasting disease currently without cure. We investigated the use of the PiggyBac transposon for full-length dystrophin expression in murine mesoangioblast (MABs) progenitor cells. DMD murine MABs were transfected with transposable expression vectors for full-length dystrophin and transplanted intramuscularly or intra-arterially into mdx/SCID mice. Intra-arterial delivery indicated that the MABs could migrate to regenerating muscles to mediate dystrophin expression. Intramuscular transplantation yielded dystrophin expression in 11%-44% of myofibers in murine muscles, which remained stable for the assessed period of 5 months. The satellite cells isolated from transplanted muscles comprised a fraction of MAB-derived cells, indicating that the transfected MABs may colonize the satellite stem cell niche. Transposon integration site mapping by whole-genome sequencing indicated that 70% of the integrations were intergenic, while none was observed in an exon. Muscle resistance assessment by atomic force microscopy indicated that 80% of fibers showed elasticity properties restored to those of wild-type muscles. As measured in vivo, transplanted muscles became more resistant to fatigue. This study thus provides a proof-of-principle that PiggyBac transposon vectors may mediate full-length dystrophin expression as well as functional amelioration of the dystrophic muscles within a potential autologous cell-based therapeutic approach of DMD., (Copyright © 2018 The American Society of Gene and Cell Therapy. Published by Elsevier Inc. All rights reserved.)
- Published
- 2018
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