Ex vivo gene transfer using adenovirus-mediated full-length dystrophin delivery to dystrophic muscles

Abstract

Duchenne muscular dystrophy (DMD) is an X-linked associated proteins. A greater amount of dystrophin recessive muscle disease characterized by a lack of dysreplacement occurred in mdx muscle following transplantrophin expression. Myoblast transplantation and gene tation of mdx myoblasts isolated from a transgenic mouse therapy have the potential of restoring dystrophin, thus overexpressing dystrophin suggesting that engineering decreasing the muscle weakness associated with this disautologous myoblasts to express high amounts of dystroease. In this study we present data on the myoblast phin might be beneficial. The ex vivo approach possesses mediated ex vivo gene transfer of full-length dystrophin to attributes that make it useful for gene transfer to skeletal mdx (dystrophin deficient) mouse muscle as a model for muscle including: (1) creating a reservoir of myoblasts capautologous myoblast transfer. Both isogenic primary mdx able of regenerating and restoring dystrophin to dystrophic myoblasts and an immortalized mdx cell line were transmuscle; and (2) achieving a higher level of gene transfer duced with an adenoviral vector that has all viral coding to dystrophic muscle compared with adenovirus-mediated sequences deleted and encodes ␤-galactosidase and fulldirect gene delivery. However, as observed in direct gene length dystrophin. Subsequently, these transduced myotransfer studies, the ex vivo approach also triggers a cellublasts were injected into dystrophic mdx muscle, where the lar immune response which limits the duration of transinjected cells restored dystrophin, as well as dystrophingene expression.

Muscle-derived cell-mediated ex vivo gene therapy for urological dysfunction

“…41 However, it is clear to us that the use of the adenoviral vector as well as the bacterial ␤-galactosidase to follow the fate Gene Therapy of the injected cells will trigger an immune response in long-term experiments. 42 In summary, we have (1) demonstrated the feasibility and survival of MDC injection into the bladder wall; (2) established improved detrusor contractility with MDC injection in a bladder injury model; (3) revealed the maturity of the ␤-galactosidase expressing myofibers in the injured bladder by demonstrating the presence of neuromuscular junctions based on the accumulation of AChRs in small segments of their membrane; and (4) demonstrated the possiblity of MDC differentiating into a smooth muscle lineage when injected into the bladder wall. We hypothesize that autologous MDC injections (muscle cells harvested from and cultured for a specific incontinence patient) can be used as a nonallergenic agent to improve bladder contractility.…”

Myoblast transplantations lead to the expression of the laminin α2 chain in normal and dystrophic (dy/dy) mouse muscles

“…To avoid this problem, ex vivo gene therapy approaches should be developed, and the in vitro engineering of dystrophic myoblasts for the expression of the therapeutic ␣2 chain gene is currently under investigation. 43,44 The understanding and control of premature cell death following injection should also be beneficial to overall transplantation results. Indeed, the success of myoblast transplantation has been recently improved by manipulating the immune response [40][41][42]44,45 or the biochemical adequacy between the donor cell and the recipient muscle 42 in other mouse models.…”

High efficiency transduction of human VEGF165 into human skeletal myoblasts: in vitro studies

“…The feasibility of ex-vivo cell mediated gene transfer is being widely investigated (Pagel et al, 1995;Floyd et al, 1998). More recently, genetically engineered myoblast mediated FGF2 delivery for revasularization in a model of acute skin flap ischemia has been documented (Rinch et al, 2001).…”