Adeno-associated virus vector transduction of vascular smooth muscle cells in vivo.
Adeno-associated virus (AAV) vectors might offer solutions for restenosis and angiogenesis by transducing nondividing cells and providing long-term gene expression. We investigated the feasibility of vascular cell transduction by AAV vectors in an in vivo rabbit carotid artery model. Time course of gene expression, inflammatory reaction to the vector, and effects of varying viral titer, exposure time, and intraluminal pressures on gene expression were examined. Recombinant AAV vectors with an Rous sarcoma virus promoter and alkaline phosphatase reporter gene were injected intraluminally into transiently isolated carotid segments. Following transduction, gene expression increased significantly over 14 days and then remained stable to 28 days, the last time point examined. Medial vascular smooth muscle cells were the main cell type transduced even with an intact endothelial layer. Increasing the viral titer and intraluminal pressure both enhanced transduction efficiency to achieve a mean of 34 +/- 7% of the subintimal layer of smooth muscle cells expressing gene product. A mild inflammatory reaction, composed of T cells with only rare macrophages, with minimal intimal thickening was demonstrated in 40% of transduced vessels; inflammatory cells were not detected in sham-operated control arteries. These findings demonstrate that AAV is a promising vector for intravascular applications in coronary and peripheral vascular diseases.
Richter
M
M
Division of Cardiothoracic Surgery, Department of Surgery, University of Washington, Fred Hutchinson Cancer Research Center, Seattle, Washington, USA.
Iwata
A
A
Nyhuis
J
J
Nitta
Y
Y
Miller
A D
AD
Halbert
C L
CL
Allen
M D
MD
D000469:Alkaline Phosphatase / Q000096:biosynthesis / Q000235:genetics; D000818:Animals; D002339:Carotid Arteries / Q000378:metabolism* / Q000601:surgery / Q000821:virology; D000229:Dependovirus / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D015870:Gene Expression; D017930:Genes, Reporter; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors / Q000008:administration & dosage / Q000378:metabolism; D007269:Injections, Intra-Arterial; D008954:Models, Biological; D009131:Muscle, Smooth, Vascular / Q000378:metabolism* / Q000821:virology; D011817:Rabbits; D013525:Surgical Instruments; D014162:Transfection; D014652:Vascular Diseases / Q000628:therapy*
D000469:Alkaline Phosphatase