BACKGROUND: Extracellular ATP and ADP may be important mediators of vascular inflammation and thrombosis. Nucleoside triphosphate diphosphohydrolase (NTPDase or CD39) is a vascular ectoenzyme that hydrolyses ATP and ADP; however, this activity is lost during reperfusion injury. We show that the supplementation of NTPDase activity within xenograft vasculature using CD39 recombinant adenoviruses (AdCD39) has protective effects in vivo. METHODS: Recombinant adenoviruses containing human CD39 or beta-galactosidase (Adbeta-gal) encoding genes were constructed. Hartley guinea pig coronary arteries were perfused ex vivo with University of Wisconsin solution containing 10(9) plaque-forming units of the recombinant adenovirus. Infected grafts were then implanted in the abdomen of complement depleted Lewis rats. RESULTS: NTPDase activities decreased in all grafts within the first 24 hr and subsequently recovered only in those hearts infected with AdCD39. Immunohistological examination of AdCD39-infected grafts confirmed successful CD39 gene transfer into the endocardium and macrovasculature. Expression of CD39 modestly prolonged graft survival (90.2+/-5.4 hr, mean+/-SD, n=5) when compared with Adbeta-gal-infected grafts (67.4+/-5.4 hr, P<0.005) and perfusion controls (66.4+/-5.2 hr; P<0.005). CONCLUSIONS: Recombinant adenoviral infection can induce expression of CD39 within cardiac xenografts and provide survival benefits in vivo. Our data show that ex vivo infection by recombinant adenovirus vectors can result in vascular expression of a potential therapeutic agent.
BACKGROUND: Extracellular ATP and ADP may be important mediators of vascular inflammation and thrombosis. Nucleoside triphosphate diphosphohydrolase (NTPDase or CD39) is a vascular ectoenzyme that hydrolyses ATP and ADP; however, this activity is lost during reperfusion injury. We show that the supplementation of NTPDase activity within xenograft vasculature using CD39 recombinant adenoviruses (AdCD39) has protective effects in vivo. METHODS: Recombinant adenoviruses containing humanCD39 or beta-galactosidase (Adbeta-gal) encoding genes were constructed. Hartley guinea pig coronary arteries were perfused ex vivo with University of Wisconsin solution containing 10(9) plaque-forming units of the recombinant adenovirus. Infected grafts were then implanted in the abdomen of complement depleted Lewis rats. RESULTS: NTPDase activities decreased in all grafts within the first 24 hr and subsequently recovered only in those hearts infected with AdCD39. Immunohistological examination of AdCD39-infected grafts confirmed successful CD39 gene transfer into the endocardium and macrovasculature. Expression of CD39 modestly prolonged graft survival (90.2+/-5.4 hr, mean+/-SD, n=5) when compared with Adbeta-gal-infected grafts (67.4+/-5.4 hr, P<0.005) and perfusion controls (66.4+/-5.2 hr; P<0.005). CONCLUSIONS: Recombinant adenoviral infection can induce expression of CD39 within cardiac xenografts and provide survival benefits in vivo. Our data show that ex vivo infection by recombinant adenovirus vectors can result in vascular expression of a potential therapeutic agent.
Authors: Maithili Sashindranath; Karen M Dwyer; Shala Dezfouli; Carly Selan; Sandra Crikis; Bo Lu; Yuping Yuan; Michael J Hickey; Karlheinz Peter; Simon C Robson; Peter J Cowan; Harshal H Nandurkar Journal: Purinergic Signal Date: 2017-03-25 Impact factor: 3.765
Authors: Martin Wijkstrom; Rita Bottino; Hayoto Iwase; Hidetaka Hara; Burcin Ekser; Dirk van der Windt; Cassandra Long; Frederico G S Toledo; Carol J Phelps; Massimo Trucco; David K C Cooper; David Ayares Journal: Xenotransplantation Date: 2014-11-10 Impact factor: 3.907
Authors: Holger K Eltzschig; David Köhler; Tobias Eckle; Tianqing Kong; Simon C Robson; Sean P Colgan Journal: Blood Date: 2008-09-23 Impact factor: 22.113
Authors: Karen M Dwyer; Simon C Robson; Harshal H Nandurkar; Duncan J Campbell; Hilton Gock; Lisa J Murray-Segal; Nella Fisicaro; Tharun B Mysore; Elzbieta Kaczmarek; Peter J Cowan; Anthony J F d'Apice Journal: J Clin Invest Date: 2004-05 Impact factor: 14.808
Authors: K Koziak; M Bojakowska; S C Robson; K Bojakowski; J Soin; E Csizmadia; P Religa; Z Gaciong; E Kaczmarek Journal: J Thromb Haemost Date: 2008-07-01 Impact factor: 5.824