Adenovirus-mediated transfer of a dominant-negative H-ras suppresses neointimal formation in balloon-injured arteries in vivo.
Abnormal migration and proliferation of arterial smooth muscle cells may be a central event in inflammatory proliferative arterial diseases such as atherosclerosis and restenosis after angioplasty. The proto-oncogene c-H-ras is considered to be a key transducer in various growth-signaling events. We constructed an adenoviral vector (AdexCAHRasY57) expressing a potent dominant-negative mutated form of c-H-ras in which tyrosine replaces aspartic acid at residue 57. Infection of smooth muscle cells with AdexCAHRasY57 produced a large quantity of H-ras-p21, completely inhibited serum-stimulated activation of mitogen-activated protein kinase, and abolished the DNA synthesis in response to serum mitogens. However, a surge of intracellular Ca2+ concentration in response to platelet-derived growth factor was not affected, suggesting that some cellular functions were preserved. When we applied AdexCAHRasY57 into balloon-injured rat carotid arteries from inside the lumen, neointimal formation was significantly reduced (neointima/media ratio: 0.28) compared with that (1.50) in arteries treated with either injury alone or injury and infection with a control adenovirus, AdexCALacZ, expressing bacterial beta-galactosidase. Our results suggest that adenovirus-mediated arterial transfer of dominant-negative H-ras may be a practical form of effective molecular intervention for proliferative arterial diseases.
Ueno
H
H
Department of Cardiology, Kyushu University School of Medicine, Fukuoka, Japan. ueno@cardiol.med.kyushu-u.ac.jp
Yamamoto
H
H
Ito
S
S
Li
J J
JJ
Takeshita
A
A
D000256:Adenoviridae / Q000235:genetics*; D000800:Angioplasty, Balloon / Q000009:adverse effects*; D000818:Animals; D001158:Arteries / Q000293:injuries* / Q000473:pathology; D001224:Aspartic Acid; D002118:Calcium / Q000378:metabolism; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D002339:Carotid Arteries / Q000473:pathology; D020212:Carotid Artery Injuries; D002470:Cell Survival; D002478:Cells, Cultured; D004247:DNA / Q000096:biosynthesis; D004789:Enzyme Activation; D018014:Gene Transfer Techniques; D011905:Genes, ras; D005822:Genetic Vectors; D009154:Mutation; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D016133:Polymerase Chain Reaction; D051381:Rats; D014443:Tyrosine
D010982:Platelet-Derived Growth Factor; D001224:Aspartic Acid; D014443:Tyrosine; D004247:DNA; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D002118:Calcium