title: Successful transfection of genes using AAV-2/9 vector in swine coronary and peripheral arteries.
abstract: BACKGROUND
  Gene therapy has attracted attention for its potential to treat several cardiovascular diseases. The use of adeno-associated viral (AAV) vectors to facilitate therapeutic gene transfer to suppress intimal hyperplasia is a promising concept. The objective of this study was to analyze the in vivo transduction of a novel recombinant AAV-2/9 vector with SM22α promoter, containing β-galactosidase gene (LacZ) or green fluorescent protein (GFP) as reporter genes, to the medial layer smooth muscle cells (SMCs) of swine coronary and peripheral arteries.
  
  
  METHODS
  The AAV-2/9 vector containing SM22α (1 × 10(13) pfu) were administered into carotid/femoral/coronary arteries of domestic swine using irrigating balloon catheter-based gene delivery. Following gene transfer, cryosections of arteries were processed for X-Gal and GFP analysis. Fluorescence microscopy and Western blotting were done to analyze the GFP expression in the SMCs.
  
  
  RESULTS
  LacZ mRNA expression was visualized in the medial layer 7 d after vector administration. The GFP expression was detected at day 7 and lasted for at least 2 mo showing the longer-lasting expression of the AAV-2/9 vector. Control arteries did not show any expression of GFP or LacZ. There was no significant effect of AAV-2/9 viral transduction on serum amylase, fibrinogen, and serum CRP levels.
  
  
  CONCLUSIONS
  These finding support the use of AAV-2/9 as a vector to effectively transduce a gene in SMCs of coronary and peripheral arteries without causing inflammation.
authors.affiliation: Center for Clinical and Translational Science, Creighton University School of Medicine, Omaha, Nebraska 68178, USA.
authors.forename: Divya
authors.initials: D
authors.lastname: Pankajakshan
authors.forename: Toluwalope O
authors.initials: TO
authors.lastname: Makinde
authors.forename: Rohit
authors.initials: R
authors.lastname: Gaurav
authors.forename: Michael
authors.initials: M
authors.lastname: Del Core
authors.forename: George
authors.initials: G
authors.lastname: Hatzoudis
authors.forename: Iraklis
authors.initials: I
authors.lastname: Pipinos
authors.forename: Devendra K
authors.initials: DK
authors.lastname: Agrawal
mesh_terms: D000818:Animals; D001158:Arteries / Q000378:metabolism; D002339:Carotid Arteries; D003331:Coronary Vessels / Q000378:metabolism; D000229:Dependovirus; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006965:Hyperplasia; D007763:Lac Operon; D008840:Microfilament Proteins / Q000378:metabolism; D009124:Muscle Proteins / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000276:immunology; D032389:Myocytes, Smooth Muscle / Q000276:immunology; D010957:Plasmids; D013552:Swine; D014162:Transfection; D017539:Tunica Intima / Q000473:pathology*
chemical_list: D008840:Microfilament Proteins; D009124:Muscle Proteins; C051615:transgelin; D049452:Green Fluorescent Proteins