Gene Therapy for Pyruvate Dehydrogenase E1[alpha] Deficiency Using Recombinant Adeno-Associated Virus 2 (rAAV2) Vectors

• Published in: Molecular therapy Vol. 6; no. 3; p. 394
• Main Authors: Owen, Renius, Mandel, Ronald J, Ammini, Chandramohan V, Conlon, Thomas J, Kerr, Douglas S, Stacpoole, Peter W, Flotte, Terence R
• Format: Journal Article
• Language: English
• Published: Milwaukee Elsevier Limited 01.09.2002
• Subjects: Acidosis; Biochemistry; Brain; Cytomegalovirus; Dehydrogenases; Efficiency; Fibroblasts; Flow cytometry; Gene therapy; Genetics; Infections; Localization; Metabolism; Microscopy; Mitochondria; Mitochondrial DNA; Patients; Pediatrics; Phosphorylation; Proteins; Vectors (Biology)
• ISSN: 1525-0016; 1525-0024
• DOI: 10.1006/mthe.2002.0683

To determine the feasibility of gene transfer to correct defects in the E1α subunit of the pyruvate dehydrogenase (PDH) complex (PDC), we constructed rAAV vectors that expressed PDH E1α, either alone or with a green fluorescent protein tag, from a hybrid cytomegalovirus (CMV) enhancer/chicken β-actin (CB) promoter. These vectors were functional in vitro, as judged by increased expression of mRNA in vector-transduced deficient cell lines and correction of the biochemical defect in PDH activity in these cells. Approximately 30% of wild-type levels of PDH activity were restored under conditions with which only about 15% of cells were transduced. These same vectors were then used in vivo to transduce neurons within the rat striatum. Gene transfer, expression, and translocation into mitochondria were observed, without any obvious untoward effects. In vivo vector-mediated PDH expression persisted for at least 1 year after injection, indicating the stability of gene transfer. These studies provide the basis for future efforts to develop a recombinant AAV (rAAV)-based gene therapy approach for the correction of PDC deficiency.