Gene transfer of IkappaBalpha limits infarct size in a mouse model of myocardial ischemia-reperfusion injury.

Abstract

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.

• Authors: Francesco Squadrito, Barbara Deodato, Giovanni Squadrito, Paolo Seminara, Maria Passaniti, Francesco S Venuti, Mauro Giacca, Letteria Minutoli, Elena B Adamo, Maria Bellomo, Rolando Marini, Mariarosaria Galeano, Herbert Marini, Domenica Altavilla
• Journal: Laboratory investigation; a journal of technical methods and pathology (Lab Invest) Vol. 83 Issue 8 Pg. 1097-104 (Aug 2003) ISSN: 0023-6837 [Print] United States
• PMID: 12920239 (Publication Type: Journal Article)

Chemical References

• DNA Primers
• I-kappa B Proteins
• NF-kappa B
• Nfkbia protein, mouse
• RNA, Messenger
• Intercellular Adhesion Molecule-1
• NF-KappaB Inhibitor alpha
• beta-Galactosidase
• Pancreatic Elastase

Topics

• Adenoviridae (genetics)
• Animals
• Blotting, Western
• DNA Primers (chemistry)
• Gene Transfer Techniques
• Genetic Therapy
• Heart Ventricles (metabolism, pathology)
• I-kappa B Proteins (genetics)
• Intercellular Adhesion Molecule-1 (genetics, metabolism)
• Male
• Mice
• Mice, Inbred C57BL
• Myocardial Infarction (genetics, pathology, prevention & control)
• Myocardial Reperfusion Injury
• NF-KappaB Inhibitor alpha
• NF-kappa B (metabolism)
• Pancreatic Elastase (metabolism)
• RNA, Messenger (metabolism)
• Transduction, Genetic (methods)
• beta-Galactosidase (metabolism)