title: Systemic correction of a fatty acid oxidation defect by intramuscular injection of a recombinant adeno-associated virus vector.
abstract: Mitochondrial beta-oxidation of fatty acids is required to meet physiologic energy requirements during illness and periods of fasting or physiologic stress, and is most active in liver and striated muscle. Acyl-CoA dehydrogenases of varying chain-length specificities represent the first step in the mitochondria for each round of beta-oxidation, each of which removes two-carbon units as acetyl-CoA for entry into the tricarboxylic acid cycle. We have used recombinant adeno-associated virus (rAAV) vectors expressing short-chain acyl-CoA dehydrogenase (SCAD) to correct the accumulation of fatty acyl-CoA intermediates in deficient cell lines. The rAAV-SCAD vector was then packaged into either rAAV serotype 1 or 2 capsids and injected intramuscularly into SCAD-deficient mice. A systemic effect was observed as judged by restoration of circulating butyryl- carnitine levels to normal. Total lipid content at the injection site was also decreased as demonstrated by noninvasive magnetic resonance spectroscopy (MRS). SCAD enzyme activity in the injected muscle was found at necropsy to be above the normal control mouse level. This study is the first to demonstrate the systemic correction of a fatty acid oxidation disorder with rAAV and the utility of MRS as a noninvasive method to monitor SCAD correction after in vivo gene therapy.
authors.affiliation: Department of Pediatrics, Powell Gene Therapy Center, University of Florida, Gainesville, FL 32610, and Department of Pediatrics, Children's Hospital of Pittsburgh, PA, USA.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Conlon
authors.forename: Glenn
authors.initials: G
authors.lastname: Walter
authors.forename: Renius
authors.initials: R
authors.lastname: Owen
authors.forename: Travis
authors.initials: T
authors.lastname: Cossette
authors.forename: Kirsten
authors.initials: K
authors.lastname: Erger
authors.forename: Greg
authors.initials: G
authors.lastname: Gutierrez
authors.forename: Eric
authors.initials: E
authors.lastname: Goetzman
authors.forename: Dietrich
authors.initials: D
authors.lastname: Matern
authors.forename: Jerry
authors.initials: J
authors.lastname: Vockley
authors.forename: Terence R
authors.initials: TR
authors.lastname: Flotte
mesh_terms: D000818:Animals; D044926:Butyryl-CoA Dehydrogenase / Q000172:deficiency / Q000235:genetics / Q000378:metabolism; D002331:Carnitine / Q000031:analogs & derivatives / Q000032:analysis / Q000097:blood; D002460:Cell Line; D004274:DNA, Recombinant; D000229:Dependovirus / Q000201:enzymology / Q000502:physiology*; D005227:Fatty Acids / Q000032:analysis / Q000378:metabolism*; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000502:physiology*; D006801:Humans; D007273:Injections, Intramuscular; D008052:Lipid Metabolism, Inborn Errors / Q000628:therapy*; D013058:Mass Spectrometry / Q000379:methods; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008928:Mitochondria / Q000378:metabolism; D009132:Muscles / Q000737:chemistry / Q000201:enzymology; D010084:Oxidation-Reduction; D015203:Reproducibility of Results; D014161:Transduction, Genetic
chemical_list: D004274:DNA, Recombinant; D005227:Fatty Acids; C427065:butyrylcarnitine; D044926:Butyryl-CoA Dehydrogenase; D002331:Carnitine