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Retroviral hybrid LTR vector strategy: functional analysis of LTR elements and generation of endothelial cell specificity

Gene Therapy volume 11, pages 775–783 (2004)Cite this article

Abstract

Transcriptional targeting is an important aspect of developing gene therapy vectors in order to restrict transgene expression to selected target cells. One approach, when using retroviral vectors, is to replace viral transcriptional control elements within the long terminal repeat (LTR) with sequences imparting the desired specificity. We have developed such hybrid LTR retroviruses, incorporating sequences from each of the human promoters for flt-1, ICAM-2 and KDR, as part of our antivascular cancer gene therapy strategy targeting tumour endothelial cells. The chosen fragments were used to replace the enhancer or combined enhancer and proximal promoter regions of the viral LTR. All showed activity in primary human breast microvascular endothelial cells, with viruses incorporating ICAM-2 sequences exhibiting the greatest specificity versus nonendothelial cells in vitro and a marked alteration of specificity towards endothelial cells in a subcutaneous xenograft model in vivo. Moreover, our study documents the effect of enhancer and/or proximal promoter deletion on LTR activity and reports that differential dependence in different cell lines can give the false impression of specificity if experiments are not adequately controlled. This finding also has implications for other retroviral vector designs seeking to provide transcriptional specificity and for their safety with respect to prevention of gene activation at sites of proviral integration.

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Acknowledgements

This work was supported by the Medical Research Council. We are grateful to the Institute of Cancer Research Biological Services Unit for animal husbandry and to S Clinton for preparing the sections. In addition, we are grateful to F-L Cosset for providing the TEFLY A8 packaging cell line; RC Gallo for the KS Y-1 cell line; the Centers for Disease Control and Prevention, Atlanta for the HMEC-1 cell line; and C Clarke and M O'Hare for the primary breast microvascular endothelial cells.

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  1. Institute of Cancer Research, Fulham Road, London, UK

    T B Richardson, J Kaspers & C D Porter

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Richardson, T., Kaspers, J. & Porter, C. Retroviral hybrid LTR vector strategy: functional analysis of LTR elements and generation of endothelial cell specificity. Gene Ther 11, 775–783 (2004). https://doi.org/10.1038/sj.gt.3302220

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