| Project/Area Number |
14370405
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Thoracic surgery
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| Research Institution | Tokyo Medical and Dental University |
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Principal Investigator |
TABUCHI Noriyuki Tokyo Medical & Dental University, Medical Hospital, Lecturer, 医学部附属病院, 講師 (90282748)
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| Co-Investigator(Kenkyū-buntansha) |
KOYAMA Takatoshi Tokyo Medical and Dental University, Graduate School of Health Sciences, Associate Professor, 大学院・保健衛生学研究科, 助教授 (20234916)
SHICHIRI Masayoshi Tokyo Medical and Dental University, Medical Hospital, Associate Professor, 医学部附属病院, 助教授 (10206097)
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| Project Period (FY) |
2002 – 2003
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| Project Status |
Completed (Fiscal Year 2003)
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| Budget Amount *help |
¥11,200,000 (Direct Cost: ¥11,200,000)
Fiscal Year 2003: ¥5,000,000 (Direct Cost: ¥5,000,000)
Fiscal Year 2002: ¥6,200,000 (Direct Cost: ¥6,200,000)
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| Keywords | gene therapy / vein graft / thrombomodulin / cornary bypass graftiing surgery / ACバイパス手術 / トロンボジュリン |
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| Research Abstract |
Immediate loss of thrombomodulin activity in the endothelium of vein grafts has been demonstrated during 90 minutes exposure to arterial cfrculation this loss of activity is ascribed as an important cause of early thrombosis. Conventional ex-vivo gene transfection after vein harvest cannot cover this acute period immediately after implantation. We have established a highly efficient noirviral gene therapy protocol utilizing modified transferrin receptor-facilitated gene transfer. Abdomen of SD rat was opened and cationic liposome : transfeitin:thrombomodulin gene complexes or the vector without DNAs were applied to the inferior vena cava of rats while blood flow was reduced by proximal and distal clamping. After 2 days, the transfected veins were harvested and thrombomodulin expression and thromboresistance properties determined before and after exposure to an artificial circuit. By transfection of the thrombomodulin gene in -IVCs, the generation capacity of activated protein C in venous endothelium increased three-fold compared with veins treated with vector alone (p<0.01). Under simulated arterial circulation, perfusion of veins treated with vector alone decreased thrombomodulin activity to 36% of preperfused levels (p<0.01), whereas transfected grafts preserved the activity at normal vein endothelium levels even after perfusion. Consequently, the increase in endothelial thrombin activity induced by simulated arterial circulation was markedly attenuated in transfected veins (p<O.01), while immunohistochemistry confirmed the preservation of endothelial lining. The present results has shown the high efficiency of our protocol of in-vivo gene therapy, and indicated the feasibility of this pre-operative gene therapy to prevent the loss of thromboresistance shortly after the implantation.
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