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Towards hematopoietic stem cell-mediated protection against infection with human immunodeficiency virus

Gene Therapy volume 13, pages 1037–1047 (2006)Cite this article

Abstract

The failure of pharmacological approaches to cure infection with the human immunodeficiency virus (HIV) has renewed the interest in gene-based therapies. Among the various strategies that are currently explored, the blockade of HIV entry into susceptible T cells and macrophages promises to be the most powerful intervention. For long-term protection of both of these lineages, genetic modification of hematopoietic stem cells (HSCs) would be required. Here, we tested whether HSCs and their progeny can be modified to express therapeutic levels of M87o, a gammaretroviral vector encoding an artificial transmembrane molecule that blocks fusion-mediated uptake of HIV. In serial murine bone marrow transplantations, efficient and multilineage expression of M87o was observed for more than 1 year (range 37–75% of mononuclear cells), without signs of toxicity related to the transmembrane molecule. To allow enrichment of M87o-modified HSCs after transplant, we constructed vectors coexpressing the P140K mutant of O6-methylguanine-DNA-methyltransferase (MGMT-P140K). This clinically relevant selection marker mediates a survival advantage in HSCs if exposed to combinations of methylguanine-methyltransferase (MGMT) inhibitors and alkylating agents. A bicistronic vector mediated sufficient expression of both M87o and MGMT to confer a selective survival advantage in the presence of HIV and alkylating agents, respectively. These data encourage further investigations in large animal models and clinical trials.

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Acknowledgements

This work was supported by grants of the European Union (MCFI-2001-51101) and Vision7 GmbH. Work at the Paterson Institute was supported by Cancer Research-UK. We are grateful to Cornelia Rudolph for assistance with fluorescence microscopy, and to Norbert Dinauer for summarizing data of the M87o bone marrow transplantation study. We thank Maimona Id and Sabine Knöß for technical assistance. The monoclonal antibody 2F5 was kindly provided by H Katinger, Vienna. Felix Hermann was supported by the European Commission project TRIoH LSHG-CT-2003-503480.

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Author notes

  1. A Schambach and B Schiedlmeier: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Hematology, Hemostaseology and Oncology, Hannover Medical School, Hannover, Germany

    A Schambach, B Schiedlmeier, M Verstegen, Z Li, J Bohne & C Baum

  2. Department of Pediatric Hematology and Oncology, Hannover Medical School, Hannover, Germany

    A Schambach

  3. EUFETS AG, Idar-Oberstein, Hannover, Germany

    K Kühlcke

  4. Department of Hematology, Erasmus, MC, The Netherlands

    M Verstegen

  5. Cancer Research-UK Carcinogenesis Group, Paterson Institute for Cancer Research, Manchester, UK

    G P Margison

  6. Institute of Molecular Pathology, Hannover Medical School, Hannover, Germany

    K Kamino

  7. Fresenius Biotech, Bad Homburg, Germany

    A Alexandrov

  8. Georg-Speyer-Haus, Frankfurt, Germany

    F G Hermann & D von Laer

  9. Division of Experimental Hematology, Cincinnati Children's Hospital Medical Center, Cincinnati, OH, USA

    C Baum

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  1. A Schambach
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Correspondence to D von Laer or C Baum.

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Supplementary Information accompanies the paper on the Gene Therapy website (http://www.nature.com/gt).

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Schambach, A., Schiedlmeier, B., Kühlcke, K. et al. Towards hematopoietic stem cell-mediated protection against infection with human immunodeficiency virus. Gene Ther 13, 1037–1047 (2006). https://doi.org/10.1038/sj.gt.3302755

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