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Foamy combinatorial anti-HIV vectors with MGMTP140K potently inhibit HIV-1 and SHIV replication and mediate selection in vivo

Gene Therapy volume 17, pages 37–49 (2010)Cite this article

Abstract

Highly active antiretroviral therapy has greatly reduced the morbidity and mortality from human immunodeficiency virus (HIV) infection, but AIDS continues to be a serious health problem worldwide. Despite enormous efforts to develop a vaccine, there is still no cure, and alternative approaches including gene therapy should be explored. In this study we developed and compared combinatorial foamy virus (FV) anti-HIV vectors that also express a mutant methylguanine methyltransferase (MGMTP140K) transgene to increase the percentage of gene-modified cells after transplantation. These FV vectors inhibit replication of HIV-1 and also the simian immunodeficiency virus/HIV-1 (SHIV) chimera that can be used in monkey AIDS gene therapy studies. We identified a combinatorial FV vector that expresses 3 anti-HIV transgenes and inhibits viral replication by over 4 logs in a viral challenge assay. This FV anti-HIV vector expresses an HIV fusion inhibitor and two short hairpin RNAs (shRNAs) targeted to HIV-1 tat and rev, and can be produced at high titer (3.8 × 107 transducing units ml−1) using improved helper plasmids suitable for clinical use. Using a competitive repopulation assay, we show that human CD34+ cells transduced with this combinatorial FV vector efficiently engraft in a mouse xenotransplantation model, and that the percentage of transduced repopulating cells can be increased after transplantation.

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Acknowledgements

This work was supported in part by grants AI063959, AI061839, DK077806, DK56465 and AI42552 from the National Institutes of Health, Bethesda, MD, USA. HPK is a Markey Molecular Medicine Investigator. We thank Tulin Okbinoglu, David Dickerson, Christina Ironside and John Ngo for technical assistance and Martin Wohlfahrt for advice and assistance with the detection of siRNA expression. We also acknowledge the assistance of Bonnie Larson and Helen Crawford in preparing this paper.

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Authors and Affiliations

  1. Clinical Research Division, Fred Hutchinson Cancer Research Center, University of Washington, Seattle, WA, USA

    H-P Kiem, R A Wu & G D Trobridge

  2. Department of Medicine, University of Washington, Seattle, WA, USA

    H-P Kiem & G D Trobridge

  3. Department of Pathology, University of Washington, Seattle, WA, USA

    H-P Kiem

  4. Department of Molecular Biology, Beckman Research Institute of City of Hope, Duarte, CA, USA

    G Sun & J J Rossi

  5. Georg-Speyer-Haus, Institute for Biomedical Research, Frankfurt, Germany

    D von Laer

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  1. H-P Kiem
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  2. R A Wu
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Correspondence to H-P Kiem or G D Trobridge.

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Kiem, HP., Wu, R., Sun, G. et al. Foamy combinatorial anti-HIV vectors with MGMTP140K potently inhibit HIV-1 and SHIV replication and mediate selection in vivo. Gene Ther 17, 37–49 (2010). https://doi.org/10.1038/gt.2009.118

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