Adenovirus-mediated transfer of human placental ectonucleoside triphosphate diphosphohydrolase to vascular smooth muscle cells suppresses platelet aggregation in vitro and arterial thrombus formation in vivo.
BACKGROUND
Platelet-rich thrombus formation is a critical event in the onset of cardiovascular disease. Because ADP plays a significant role in platelet aggregation, its metabolism is important in the regulation of platelet activation and recruitment. Ectonucleoside triphosphate diphosphohydrolase (E-NTPDase) is a key enzyme involved in vascular ADP metabolism. We recently isolated 2 isoforms of E-NTPDase from the human placenta. The present study examined whether these isoforms suppress platelet aggregation and thrombus formation after adenovirus-mediated gene transfer to vascular smooth muscle cells (SMCs).


RESULTS
We constructed adenovirus vectors expressing human placental E-NTPDase isoforms I (AdPlac I) and II (AdPlac II) or bacterial beta-galactosidase (AdLacZ). Vascular SMCs infected with AdPlac I expressed significant NTPDase activity and inhibited the platelet aggregation induced by ADP and collagen in vitro. In contrast, SMCs infected with AdPlac II and AdLacZ did not exert antiplatelet effects. To investigate the antithrombotic and antiproliferative effects of placental E-NTPDase isoform I in vivo, we generated thrombosis in rat carotid arteries by systemically administered rose Bengal and transluminal green light 5 days after gene transfer and examined neointimal growth 3 weeks after thrombus formation. Blood flow in AdLacZ-infected arteries rapidly deteriorated and vanished within 96+/-18 seconds of occlusive thrombus formation. In contrast, blood flow in AdPlac I-infected arteries was preserved for at least 10 minutes during irradiation. In addition, thrombus formation and subsequent neointimal growth were obviously suppressed.


CONCLUSIONS
The local expression of placental E-NTPDase in injured arteries might prevent arterial thrombosis and subsequent neointimal growth.
Furukoji
Eiji
E
Department of Radiology, Miyazaki Medical College, University of Miyazaki, Kiyotake, Miyazaki, Japan.
Matsumoto
Masanori
M
Yamashita
Atsushi
A
Yagi
Hideo
H
Sakurai
Yoshihiko
Y
Marutsuka
Kousuke
K
Hatakeyama
Kinta
K
Morishita
Kazuhiro
K
Fujimura
Yoshihiro
Y
Tamura
Shozo
S
Asada
Yujiro
Y
D000251:Adenosine Triphosphatases / Q000096:biosynthesis / Q000235:genetics*; D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis / Q000235:genetics*; D001013:Aorta, Thoracic; D001081:Apyrase / Q000096:biosynthesis / Q000235:genetics*; D001777:Blood Coagulation; D002339:Carotid Arteries / Q000601:surgery; D002478:Cells, Cultured; D006801:Humans; D008026:Ligation; D008297:Male; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000201:enzymology* / Q000821:virology; D032389:Myocytes, Smooth Muscle / Q000201:enzymology / Q000821:virology; D009389:Neovascularization, Pathologic; D010920:Placenta / Q000201:enzymology*; D010974:Platelet Aggregation / Q000502:physiology / Q000528:radiation effects; D010979:Platelet Function Tests; D051381:Rats; D017207:Rats, Sprague-Dawley; D013927:Thrombosis / Q000517:prevention & control; D014161:Transduction, Genetic / Q000379:methods*; D017539:Tunica Intima / Q000378:metabolism
D015703:Antigens, CD; D000251:Adenosine Triphosphatases; C027201:ectoATPase; D001081:Apyrase; C067869:CD39 antigen