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Recombinant adeno-associated virus vector-transduced vascular endothelial cells express the thrombomodulin transgene under the regulation of enhanced plasminogen activator inhibitor-1 promoter

Gene Therapy volume 8, pages 1690–1697 (2001)Cite this article

Abstract

We were able to facilitate plasminogen activator inhibitor 1 (PAI-1) promoter activity approximately by 14-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 TGFβ1 and TNFα stimulation. The characteristics of the enhanced PAI-1 promoter are 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 between 600- and 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 could also be transduced by the same rAAV vector in vivo. Transduction 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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Acknowledgements

This work is supported by Grants-in-aid for Scientific Research 12670687 to JM from the Ministry of Education.

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Authors and Affiliations

  1. Division of Cell and Molecular Medicine, Center for Molecular Medicine, Jichi Medical School, Tochigi-ken, Japan

    J Mimuro, S Muramatsu, K Mori, J Kikuchi, S Madoiwa & Y Sakata

  2. Division of Genetic Therapeutics, Center for Molecular Medicine, Jichi Medical School, Tochigi-ken, Japan

    S Muramatsu, M Urabe & K Ozawa

  3. Division of Developmental Biotechnology, Center for Molecular Medicine, Jichi Medical School, Tochigi-ken, Japan

    Y Hakamada

  4. Department of Hematology, Jichi Medical School, Tochigi-ken, Japan

    J Mimuro, S Madoiwa, K Ozawa & Y Sakata

  5. Department of Neurology, Jichi Medical School, Tochigi-ken, Japan

    S Muramatsu

  6. Tsukuba Research Institute, Nippon Shinyaku Co Ltd, Ibraki-ken, Japan

    K Mori

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  1. J Mimuro
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Mimuro, J., Muramatsu, S., Hakamada, Y. et al. Recombinant adeno-associated virus vector-transduced vascular endothelial cells express the thrombomodulin transgene under the regulation of enhanced plasminogen activator inhibitor-1 promoter. Gene Ther 8, 1690–1697 (2001). https://doi.org/10.1038/sj.gt.3301579

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  • DOI: https://doi.org/10.1038/sj.gt.3301579

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