title: Correction of the enzymatic and functional deficits in a model of Pompe disease using adeno-associated virus vectors.
abstract: Pompe disease is a lysosomal storage disease caused by the absence of acid alpha-1,4 glucosidase (GAA). The pathophysiology of Pompe disease includes generalized myopathy of both cardiac and skeletal muscle. We sought to use recombinant adeno-associated virus (rAAV) vectors to deliver functional GAA genes in vitro and in vivo. Myotubes and fibroblasts from Pompe patients were transduced in vitro with rAAV2-GAA. At 14 days postinfection, GAA activities were at least fourfold higher than in their respective untransduced controls, with a 10-fold increase observed in GAA-deficient myotubes. BALB/c and Gaa(-/-) mice were also treated with rAAV vectors. Persistent expression of vector-derived human GAA was observed in BALB/c mice up to 6 months after treatment. In Gaa(-/-) mice, intramuscular and intramyocardial delivery of rAAV2-Gaa (carrying the mouse Gaa cDNA) resulted in near-normal enzyme activities. Skeletal muscle contractility was partially restored in the soleus muscles of treated Gaa(-/-) mice, indicating the potential for vector-mediated restoration of both enzymatic activity and muscle function. Furthermore, intramuscular treatment with a recombinant AAV serotype 1 vector (rAAV1-Gaa) led to nearly eight times normal enzymatic activity in Gaa(-/-) mice, with concomitant glycogen clearance as assessed in vitro and by proton magnetic resonance spectroscopy.
authors: Powell Gene Therapy Center, University of Florida College of Medicine, Gainesville, Florida 32610, USA.
authors: Thomas J
authors: TJ
authors: Fraites
authors: Mary R
authors: MR
authors: Schleissing
authors: R Andrew
authors: RA
authors: Shanely
authors: Glenn A
authors: GA
authors: Walter
authors: Denise A
authors: DA
authors: Cloutier
authors: Irene
authors: I
authors: Zolotukhin
authors: Daniel F
authors: DF
authors: Pauly
authors: Nina
authors: N
authors: Raben
authors: Paul H
authors: PH
authors: Plotz
authors: Scott K
authors: SK
authors: Powers
authors: Paul D
authors: PD
authors: Kessler
authors: Barry J
authors: BJ
authors: Byrne
mesh_terms: D000818:Animals; D002318:Cardiovascular Diseases / Q000378:metabolism / Q000628:therapy; D000229:Dependovirus / Q000235:genetics*; D004195:Disease Models, Animal; D005347:Fibroblasts / Q000378:metabolism; D005786:Gene Expression Regulation; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006003:Glycogen / Q000378:metabolism; D006009:Glycogen Storage Disease Type II / Q000201:enzymology / Q000235:genetics / Q000628:therapy*; D006720:Homozygote; D006801:Humans; D007124:Immunoenzyme Techniques; D007223:Infant; D016464:Lysosomal Storage Diseases / Q000378:metabolism / Q000628:therapy; D051379:Mice; D008807:Mice, Inbred BALB C; D018345:Mice, Knockout; D018482:Muscle, Skeletal / Q000503:physiopathology*; D009206:Myocardium / Q000378:metabolism; D014161:Transduction, Genetic; D000520:alpha-Glucosidases / Q000235:genetics* / Q000378:metabolism
chemical_list: D006003:Glycogen; D000520:alpha-Glucosidases