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Lentiviral vector conferring resistance to mycophenolate mofetil and sensitivity to ganciclovir for in vivo T-cell selection

Gene Therapy volume 14, pages 1549–1554 (2007)Cite this article

Abstract

Several clinical studies of gene-modified T cells have shown limited in vivo function of the cells, immunogenicity of the transgene, and lack of a selective advantage for gene-modified T cells. To address these problems, we developed a lentiviral vector (LV) that provides a selectable, proliferative advantage and potentially decreases immunogenicity for transduced T cells. The bicistronic vector expressed two genes linked with an internal ribosomal entry site. One gene is a variant of the inosine monophosphate dehydrogenase 2, inosine monophosphate dehydrogenase (IMPDHIY), conferring resistance to the immunosuppressive drug mycophenolate mofetil (MMF). The other is a suicide gene, herpes simplex virus thymidine kinase (HSV-TK), rendering proliferating cells sensitive to ablation with ganciclovir, fused to the selectable transmembrane marker ΔCD34 (ΔCD34/TK). Cells transduced with LV-ΔCD34/TK.IMPDHIY were efficiently enriched by immunomagnetic selection for CD34, proliferated in 0.5–5 μM MMF, and were killed by 0.5–25 μg ml−1 ganciclovir. We demonstrate efficient selection and killing of gene-modified cells and suggest LV-ΔCD34/TK.IMPDHIY-transduced T cells could be used to facilitate allogeneic hematopoietic cell engraftment. The expression of IMPDHIY would allow in vivo selection with MMF, and ΔCD34/TK expression would allow rapid and safe elimination of transduced T cells if graft-versus-host disease developed.

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Acknowledgements

The research was supported by NIH NCI Grants R01 CA109381, P01 CA78902, R01 DK42716 and ‘Progetto Vita Vitae’, Fondazione CRT. We are grateful to Dr John F DiPersio, Washington University, for providing the ΔCD34/HSV-TK75 fusion gene. We thank Helen Crawford and Bonnie Larson for assistance in preparation of the paper.

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Authors and Affiliations

  1. Clinical Research Division, Transplantation Biology, Fred Hutchinson Cancer Research Center, Seattle, WA, USA

    D Sangiolo, M Lesnikova, R A Nash, A Nikitine, H-P Kiem & G E Georges

  2. Department of Oncological Sciences, University of Torino Medical School, Torino, Italy

    D Sangiolo

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

    R A Nash, H-P Kiem & G E Georges

  4. Division of Cancer Immunotherapeutics and Tumor Immunology, City of Hope National Medical Center, Duarte, CA, USA

    M C Jensen

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  1. D Sangiolo
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  2. M Lesnikova
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  4. M C Jensen
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  5. A Nikitine
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  6. H-P Kiem
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  7. G E Georges
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Correspondence to G E Georges.

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Sangiolo, D., Lesnikova, M., Nash, R. et al. Lentiviral vector conferring resistance to mycophenolate mofetil and sensitivity to ganciclovir for in vivo T-cell selection. Gene Ther 14, 1549–1554 (2007). https://doi.org/10.1038/sj.gt.3303018

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