The fate of individual myoblasts after transplantation into muscles of DMD patients

Abstract

Muscle biopsies from six patients with Duchenne muscular dystrophy (DMD) participating in a myoblast transplantation clinical trial were reexamined using a fluorescence in situ hybridization (FISH)-based method. Donor nuclei were detected in all biopsies analyzed, including nine where no donor myoblasts were previously thought to be present. In three patients, more than 10% of the original number of donor cells were calculated as present 6 months after implantation. Half of the detected donor nuclei were fused into host myofibers, and of these, nearly 50% produced dystrophin. These findings demonstrate that although donor myoblasts have persisted after injection, their microenvironment influences whether they fuse and express dystrophin. Our methodology could be used for developing new approaches to improve myoblast transfer efficacy and for the analysis of future gene- and/or cell-based therapies of numerous genetic disorders.

Electrotransfer of naked DNA in the skeletal muscles of animal models of muscular dystrophies

“…Our studies confirm some previous findings obtained in other models, ie over 1500 m for ␤-gal 60,61 and about 300 m for dystrophin. 60,62 Along the same line, we show here that the domain of expression of the laminin-2 molecule is approximately 500 m. These results underline one present limitation of direct gene transfer into skeletal muscles, which is the restricted segmental diffusion of the transgenic proteins inside the transduced fibers. This may be considered an advantage in situations where the local delivery of a protein is required.…”

Skeletal myoblast transplantation for attenuation of hyperglycaemia, hyperinsulinaemia and glucose intolerance in a mouse model of type 2 diabetes mellitus

“…The basis of our cell therapy study for insulin resistance is similar to that used for the treatment of DMD [28][29][30][31]. However, DMD is a single gene defect with complete absence of production of a single protein, whereas type 2 diabetes is a polygeneic disease with potentially decreased levels of multiple proteins.…”

Adeno-associated virus vector-mediated gene transfer into dystrophin-deficient skeletal muscles evokes enhanced immune response against the transgene product

“…These include the adenoviral vector-mediated introduction of functional dystrophin, [2][3][4][5][6] naked DNA, 7 and primary myoblast 8 or stem cell 9 transplantation. Alternatively, correction of the endogenous dystrophin gene was attempted using chimeric RNA/DNA oligonucleotides 10,11 or antisense 2'-O-methyl oligoribonucleotides.…”