title: 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.
abstract: 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.
authors.affiliation: Department of Radiology, Miyazaki Medical College, University of Miyazaki, Kiyotake, Miyazaki, Japan.
authors.forename: Eiji
authors.initials: E
authors.lastname: Furukoji
authors.forename: Masanori
authors.initials: M
authors.lastname: Matsumoto
authors.forename: Atsushi
authors.initials: A
authors.lastname: Yamashita
authors.forename: Hideo
authors.initials: H
authors.lastname: Yagi
authors.forename: Yoshihiko
authors.initials: Y
authors.lastname: Sakurai
authors.forename: Kousuke
authors.initials: K
authors.lastname: Marutsuka
authors.forename: Kinta
authors.initials: K
authors.lastname: Hatakeyama
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Morishita
authors.forename: Yoshihiro
authors.initials: Y
authors.lastname: Fujimura
authors.forename: Shozo
authors.initials: S
authors.lastname: Tamura
authors.forename: Yujiro
authors.initials: Y
authors.lastname: Asada
mesh_terms: 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
chemical_list: D015703:Antigens, CD; D000251:Adenosine Triphosphatases; C027201:ectoATPase; D001081:Apyrase; C067869:CD39 antigen