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. This study reports full-length dystrophin expression from PiggyBac transposon vectors transfected in murine mesoangioblast progenitor cells (MAB) as a potential therapeutic autologous cell transplantation approach. Intramuscular MAB transplantation into dmd mice yielded stable dystrophin expression, restored a normal resistance for 80% of the myofibers, and increased muscle resistance to fatigue.

Autologous cell therapy approach for Duchenne muscular dystrophy using PiggyBac transposons and mesoangioblasts / P.S. Iyer, L.O. Mavoungou, F. Ronzoni, J. Zemla, E. Schmid-Siegert, S. Antonini, L.A. Neff, O.M. Dorchies, M. Jaconi, M. Lekka, G. Messina, N. Mermod. - In: MOLECULAR THERAPY. - ISSN 1525-0016. - 26:4(2018), pp. 1093-1108.

Autologous cell therapy approach for Duchenne muscular dystrophy using PiggyBac transposons and mesoangioblasts

S. Antonini;G. Messina;
2018

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. This study reports full-length dystrophin expression from PiggyBac transposon vectors transfected in murine mesoangioblast progenitor cells (MAB) as a potential therapeutic autologous cell transplantation approach. Intramuscular MAB transplantation into dmd mice yielded stable dystrophin expression, restored a normal resistance for 80% of the myofibers, and increased muscle resistance to fatigue.
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/2434/549907
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