Adenovirus-Mediated Transfer of the Acid α-Glucosidase Gene into Fibroblasts, Myoblasts and Myotubes from Patients with Glycogen Storage Disease Type II Leads to High Level Expression of Enzyme and Corrects Glycogen Accumulation

摘要: Glycogen storage disease type II (GSD II) is an autosomal recessive disorder caused by defects in the lysosomal acid alpha-glucosidase (GAA) gene. We investigated feasibility of using a recombinant adenovirus containing human GAA gene under control cytomegalovirus promoter (AdCMV-GAA) to correct enzyme deficiency different cultured cells from patients with infantile form GSD II. In GAA-deficient fibroblasts infected AdCMV-GAA, transduction and transcription transgene resulted de novo synthesis protein. The activity was corrected deficient level 12 times normal cells. transduced overexpressed 110 kDa precursor GAA, which secreted into culture medium taken up recipient protein correctly processed active on both artificial substrate 4-methylumbelliferyl-alpha-D-glucopyranoside (4MUG) glycogen. muscle cells, significant increase cellular level, approximately 20-fold higher than also observed after viral treatment. were able efficiently secrete GAA. Moreover, transfer normalization glycogen content clearance lysosomes, as assessed electron microscopy, differentiated myotubes. These results demonstrate phenotypic correction skeletal patient infantile-onset adenovirus. conclude that adenovirus-mediated might be suitable model system for further vivo studies delivering muscle, not only direct cell targeting but combination secretion uptake mechanisms.