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Recombinant Semliki Forest virus as a vector system for fast and selective in vivo gene delivery into balloon-injured rat aorta

Gene Therapy volume 9, pages 95–101 (2002)Cite this article

Abstract

Previously, we demonstrated that recombinant Semliki Forest virus (SFV) vector rapidly and selectively transfers genes into cultured vascular smooth muscle cells (VSMC), leaving endothelial cells (EC) unaffected. From this, we hypothesized that recombinant SFV in vivo only transfers genes into the media of balloon-injured but not intact vessel, that gene expression in VSMC is fast, and that the specificity of SFV for VSMC is caused by specific binding sites. To address these hypotheses, we studied the time course of in vivo SFV-LacZ and Ad-LacZ expression in balloon-injured rat aorta. In addition, the fusion characteristics of fluorescent pyrene-labeled SFV were explored in cultured VSMC and EC. In intact aorta, no LacZ expression was found in the intima or media at 24 h. In contrast, in denuded aorta, LacZ expression was detected in as early as 12 h after incubation. LacZ expression was predominantly present in the media. Ad-LacZ expression started after 12 h, but was predominantly present in the adventitia. Ad-LacZ expression in the media started after 72 h. In vitro transfection with SFV showed that fusion was higher and, moreover, saturable in VSMC as compared with EC, indicating the presence of specific SFV binding sites on VSMC, but not EC. From this we conclude that in vivo selectivity of SFV in balloon-injured vessels is based on the removal of the endothelium, which results in accessibility of VSMC in the media that carry specific binding sites for the SFV vector.

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Acknowledgements

We are very grateful to Hans Bartels, Alexandra Beuving, Martin Houwertjes, Gera Kamps, Bianca Meijeringh, Egbert Scholtens, Lucia van der Veen, and Jetti van Wijk, for their technical support. We thank Dr A Suurmeijer of the Department of Pathology, University Hospital Groningen for his useful suggestions. Further, we thank Medtronic-Bakken Research Maastricht, Inex Pharmaceuticals (grant to J Wilschut), and the Netherlands Heart Foundation (ICIN grant No. 295-0019 to Dr RA Tio) for their financial support.

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Authors and Affiliations

  1. Department of Clinical Pharmacology, Groningen University Institute for Drug Exploration (GUIDE), University of Groningen, The Netherlands

    A J M Roks, R H Henning, H Buikema, D de Zeeuw & W H van Gilst

  2. Department of Cardiology, University of Groningen, The Netherlands

    Y M Pinto & R A Tio

  3. Department of Medical Microbiology, Molecular Virology Section, University of Groningen, The Netherlands

    M J J Kraak & J Wilschut

  4. Department of Therapeutic Gene Modulation, University of Groningen, The Netherlands

    H J Haisma

Authors
  1. A J M Roks
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  2. R H Henning
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  3. H Buikema
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  4. Y M Pinto
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  5. M J J Kraak
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  6. R A Tio
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Roks, A., Henning, R., Buikema, H. et al. Recombinant Semliki Forest virus as a vector system for fast and selective in vivo gene delivery into balloon-injured rat aorta. Gene Ther 9, 95–101 (2002). https://doi.org/10.1038/sj.gt.3301632

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