Perivascular gene transfer of NADPH oxidase inhibitor suppresses angioplasty-induced neointimal proliferation of rat carotid artery.
Vascular stretch induces NADPH oxidase-derived superoxide anion (O2-), which has been implicated in hypertrophy and cell proliferation. We hypothesized that targeted delivery of an NADPH oxidase inhibitor to the adventitia would reduce stretch-induced vascular O2- and attenuate neointima formation. We designed a novel replication-deficient adenovirus containing a fibroblast-active promoter driving expression of NADPH oxidase inhibitory sequence gp91ds (Ad-PDGFbetaR-gp91ds/eGFP). 1) We characterized the specificity of this promoter using pPDGFbetaR-luciferase by showing induction of luciferase in cultured rat aortic fibroblasts but not in vascular smooth muscle cells. 2) Using RT-PCR, we observed expression of gp91ds and the reporter gene in fibroblasts after infection with Ad-PDGFbetaR-gp91ds/eGFP. 3) Using Ad-CMV-eGFP as a control, we delivered Ad-PDGFbetaR-gp91ds/eGFP to the adventitia of the rat common carotid artery (CCA). Immunohistochemistry confirmed localized delivery of the inhibitor to the adventitia. After CCAs were injured with an embolectomy catheter, we observed a significant increase in neointima-to-media area ratio in control CCAs, which was significantly attenuated in CCAs treated with the gp91ds-expressing virus. In a second group of rats, we detected a 10-fold increase in distension-stimulated O2-, which was significantly reduced in CCAs infected with gp91ds-expressing virus. These data demonstrate that localized adventitial delivery of an NADPH oxidase inhibitor is effective in reducing overall vascular O2- and neointima formation, suggesting that adventitial NADPH oxidase plays a functional role in development of neointimal hyperplasia.
Dourron
Hector M
HM
Division of Vascular Surgery, Henry Ford Health System, Detroit, Michigan, USA.
Jacobson
Gary M
GM
Park
James L
JL
Liu
Jianhua
J
Reddy
Daniel J
DJ
Scheel
Maria L
ML
Pagano
Patrick J
PJ
D000800:Angioplasty, Balloon / Q000009:adverse effects*; D000818:Animals; D002339:Carotid Arteries / Q000473:pathology* / Q000502:physiology; D016893:Carotid Stenosis / Q000473:pathology / Q000628:therapy*; D002478:Cells, Cultured; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D049452:Green Fluorescent Proteins / Q000235:genetics; D008156:Luciferases / Q000235:genetics; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000502:physiology; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors* / Q000235:genetics*; D011401:Promoter Regions, Genetic; D051381:Rats; D020797:Receptor, Platelet-Derived Growth Factor beta / Q000235:genetics; D013481:Superoxides / Q000378:metabolism; D017540:Tunica Media / Q000473:pathology
D013481:Superoxides; D049452:Green Fluorescent Proteins; D008156:Luciferases; D019255:NADPH Oxidases; D020797:Receptor, Platelet-Derived Growth Factor beta