| Project/Area Number |
12670687
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Circulatory organs internal medicine
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| Research Institution | Jichi Medical School |
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Principal Investigator |
MIMURO Jun Jichi Medical School, Cell and molecular medicine, Instructor, 医学部, 講師 (10221607)
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| Co-Investigator(Kenkyū-buntansha) |
MURAMATSU Shin-ichi Jichi Medical School, Cell and molecular medicine, Instructor, 医学部, 助手 (10239543)
SAKATA Yoichi Jichi Medical School, Cell and molecular medicine, Professor, 医学部, 教授 (40129028)
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| Project Period (FY) |
2000 – 2001
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| Project Status |
Completed (Fiscal Year 2001)
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| Budget Amount *help |
¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2001: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2000: ¥2,500,000 (Direct Cost: ¥2,500,000)
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| Keywords | endothelial cell / thrombosis / thrombomodulin / adeno-associated virus vector / gene therapy / plasminogen activator inhibitor 1 / AAV vector / Thrombosis |
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| Research Abstract |
We could facilitate plasminogen activator inhibitor 1 (PAI-1) promoter activity approximately by 20-fold using an enhancer element. This enhanced PAI- 1 promoter has a strong basal activity, comparable to CAG promoter activity, and has a response similar to the PAI-1 promoter with respect to TGF β1 and TNF α stimulation. The characteristic of the enhanced PAI-1 promoter is thought to be suited to timely and tissue specific expression of anticoagulant molecules in the vascular cells. Thus, we developed recombinant adeno-associated virus (rAAV) vectors using the enhanced PAI-1 promoter and were successful in transducing vascular endothelial cells to express the thrombomodulin transgene under the regulation of the enhanced PAI-1 promoter in vitro. Thromobomodulin transgene expression driven by the enhanced PAI- 1 promoter in rAAV vector-transduced cultured endothelial cells was 600〜1000-fold higher than constitutive thrombomodulin gene expression in cultured human umbilical vein endothelial cells and was up-regulated by TGFβ1 and TNFα stimulation which may down-regulate endogenous thrombomodulin gene expression in endothelial cells. The brain vascular endothelial cells of Mongolian gerbils also could be transduced by the same rAAV vector in vivo. Transition of endothelial cells by rAAV vectors to express enhanced PAI- 1 promoter-driven transgenes may be a useful gene therapy approach for vascular diseases.
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