Multigene adenoviral therapy for the attenuation of ischemia-reperfusion injury after preservation for cardiac transplantation.
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.
Abunasra
Haitham J
HJ
Heart Science Centre, Imperial College at Harefield Hospital, Harefield, United Kingdom.
Smolenski
Ryszard T
RT
Yap
John
J
Sheppard
Mary
M
O'Brien
Timothy
T
Yacoub
Magdi H
MH
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
D005697:Galactosides; D007211:Indoles; D019001:Nitric Oxide Synthase; D013482:Superoxide Dismutase; C044888:5-bromo-4-chloro-3-indolyl beta-galactoside