Gene transfer of IkappaBalpha limits infarct size in a mouse model of myocardial ischemia-reperfusion injury.
Nuclear factor-kappaB (NF-kappaB) plays a central role in myocardial ischemia-reperfusion (MI/R) injury. The inhibitory protein IkappaBalpha prevents its activation. We investigated the effects of adeno-associated viral vector-mediated IkappaBalpha gene transfer in MI/R injury. Male C57BL/6 mice were randomized to receive a recombinant adeno-associated virus (rAAV) encoding the gene for the NF-kappaB inhibitory protein IkappaBalpha (rAAV- IkappaBalpha) or the beta-galactosidase gene (a control and inert gene; rAAV-LacZ), both at a dose of 10(11) copies. Four weeks later anesthetized animals were subjected to total occlusion (45 minutes) of the left main coronary artery followed by 5 hours of reperfusion. MI/R produced a wide infarct size (IF/area-at-risk = 56 +/- 8%; IF/left ventricle = 44 +/- 5%) and tissue neutrophil infiltration, studied by means of elastase activity (area-at-risk = 2.5 +/- 0.4 micro g/gm tissue; infarct area = 2.9 +/- 0.6 micro g/gm tissue). Furthermore MI/R caused peak message for intercellular adhesion molecule-1 (ICAM-1) in the area-at-risk at 3 hours of reperfusion (1.2 +/- 0.4 relative amount of cardiac ICAM-1 mRNA). NF-kappaB activation was evident at 0.5 hours of reperfusion and reached its maximum increase at 2 hours of reperfusion. rAAV-IkappaBalpha injection reduced infarct size (IF/area-at-risk = 19 +/- 3%; IF/left ventricle = 10 +/- 2%; p < 0.001), blocked NF-kappaB activation, diminished cardiac ICAM-1 expression (0.4 +/- 0.02 relative amount of cardiac ICAM-1 mRNA; p < 0.001), and blunted leukocyte accumulation (area-at-risk = 0.6 +/- 0.05 micro g/gm tissue; infarct area = 0.4 +/- 0.02 micro g/gm tissue; p < 0.001). Our data indicate that rAAV-IkappaBalpha may be useful for MI/R gene therapy.
Squadrito
Francesco
F
Department of Clinical and Experimental Medicine and Pharmacology, University of Messina, Messina, Italy. Francesco.Squadrito@unime.it
Deodato
Barbara
B
Squadrito
Giovanni
G
Seminara
Paolo
P
Passaniti
Maria
M
Venuti
Francesco S
FS
Giacca
Mauro
M
Minutoli
Letteria
L
Adamo
Elena B
EB
Bellomo
Maria
M
Marini
Rolando
R
Galeano
Mariarosaria
M
Marini
Herbert
H
Altavilla
Domenica
D
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D015153:Blotting, Western; D017931:DNA Primers / Q000737:chemistry; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D006352:Heart Ventricles / Q000378:metabolism / Q000473:pathology; D020672:I-kappa B Proteins / Q000235:genetics*; D018799:Intercellular Adhesion Molecule-1 / Q000235:genetics / Q000378:metabolism; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D009203:Myocardial Infarction / Q000235:genetics* / Q000473:pathology / Q000517:prevention & control*; D015428:Myocardial Reperfusion Injury; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000378:metabolism; D010196:Pancreatic Elastase / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D014161:Transduction, Genetic / Q000379:methods*; D001616:beta-Galactosidase / Q000378:metabolism
D017931:DNA Primers; D020672:I-kappa B Proteins; D016328:NF-kappa B; C000606205:Nfkbia protein, mouse; D012333:RNA, Messenger; D018799:Intercellular Adhesion Molecule-1; D000072000:NF-KappaB Inhibitor alpha; D001616:beta-Galactosidase; D010196:Pancreatic Elastase