title: Multigene adenoviral therapy for the attenuation of ischemia-reperfusion injury after preservation for cardiac transplantation.
abstract: OBJECTIVE
  The protective effect of adenovirus-mediated ex vivo multigene transfer with superoxide dismutase, a free radical scavenger, and nitric oxide, a vasodilator with anti-inflammatory properties, was examined in the rat heart during experimental ischemia-reperfusion mimicking preservation for cardiac transplantation.
  
  
  METHODS
  Donor rat hearts (n = 6 per group) were perfused with solution containing adenoviral vector carrying genes for beta-galactosidase (group A), endothelial nitric oxide synthase (group B), manganese superoxide dismutase (group C), or both endothelial nitric oxide synthase and manganese superoxide dismutase (group D). Hearts were then implanted heterotopically into the abdomens of recipient rats. Four days later, transplanted hearts were collected, connected to a Langendorff perfusion apparatus, and subjected to 6 hours of ischemia followed by 1 hour of reperfusion. Cardiac function was evaluated with an intraventricular balloon at the beginning of Langendorff perfusion and after ischemia-reperfusion.
  
  
  RESULTS
  Effective gene transfection was confirmed with X-gal staining in group A hearts. Positive immunoreactivity for endothelial nitric oxide synthase, manganese superoxide dismutase, or both was present predominantly in cardiomyocytes in group B, C, and D hearts. Percentage recovery of preischemic left ventricular developed pressure was 62.1% +/- 7.36% in group A; recoveries were increased to 79.6% +/- 6.4%, 86.8% +/- 9.1%, and 79.4% +/- 6.2% in groups B, C, and D, respectively.
  
  
  CONCLUSIONS
  These results indicate that adenoviral gene transfer of manganese superoxide dismutase and endothelial nitric oxide synthase can attenuate myocardial ischemia-reperfusion injury, with the former providing the most significant protection. Combined overexpression of manganese superoxide dismutase and endothelial nitric oxide synthase did not enhance myocardial recovery any further.
authors.affiliation: Heart Science Centre, Imperial College at Harefield Hospital, Harefield, United Kingdom.
authors.forename: Haitham J
authors.initials: HJ
authors.lastname: Abunasra
authors.forename: Ryszard T
authors.initials: RT
authors.lastname: Smolenski
authors.forename: John
authors.initials: J
authors.lastname: Yap
authors.forename: Mary
authors.initials: M
authors.lastname: Sheppard
authors.forename: Timothy
authors.initials: T
authors.lastname: O'Brien
authors.forename: Magdi H
authors.initials: MH
authors.lastname: Yacoub
mesh_terms: D000256:Adenoviridae; D000818:Animals; D004195:Disease Models, Animal; D005697:Galactosides / Q000235:genetics; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D016027:Heart Transplantation; D007150:Immunohistochemistry; D007211:Indoles; D008297:Male; D015428:Myocardial Reperfusion Injury / Q000517:prevention & control*; D019001:Nitric Oxide Synthase / Q000008:administration & dosage / Q000235:genetics* / Q000378:metabolism; D009926:Organ Preservation; D051381:Rats; D017207:Rats, Sprague-Dawley; D013482:Superoxide Dismutase / Q000008:administration & dosage / Q000235:genetics* / Q000378:metabolism; D014162:Transfection
chemical_list: D005697:Galactosides; D007211:Indoles; D019001:Nitric Oxide Synthase; D013482:Superoxide Dismutase; C044888:5-bromo-4-chloro-3-indolyl beta-galactoside