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Lentiviral vectors encoding human MUC1-specific, MHC-unrestricted single-chain TCR and a fusion suicide gene: potential for universal and safe cancer immunotherapy

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Abstract

MUC1 tumor antigen is a target for immunotherapy of most human adenocarcinomas and some hematological malignancies. Expression of a MUC1-specific, MHC-unrestricted single-chain T cell receptor (scTCR) on cells of both innate and adaptive immune system through reconstitution of lethally irradiated mice by retroviral vector-transduced bone marrow cells, had been shown to effectively control the growth of MUC1+ tumors independent of their MHC type, suggesting that this receptor is a good candidate for broadly applicable gene therapy/immunotherapy. However, the translational application of this immuno-gene therapy modality was discouraged by the progressive transgene silencing in reconstituted T and B cells, as well as the potential of tumorogenesis intrinsic to oncoretroviral vectors. To overcome these problems and facilitate the future clinical use of this receptor, we have constructed a panel of novel self-inactivating lentiviral vectors (LVs) which harbor two independent internal promoters, one driving expression of the scTCR gene and the other of a fusion suicide gene, the HSV-TK–EGFP fusion gene, allowing the transduced cells to be destroyable by the pro-drug ganciclovir. Despite the large size of insert, these vectors were efficiently packaged into high titer virus that transferred the expression of transgene in both T cell lines and primary T cells. Sustained expression was maintained in a T cell line for over 4 months in vitro, suggesting its efficient resistance to transgene silencing. Both scTCR and HSV-TK–EGFP genes were functional in the transduced cells, as evidenced by their specific recognition of MUC1+ tumors and efficient eradication by ganciclovir.

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Acknowledgments

We are grateful to all the investigators around the world who generously provided us with their materials. We also thank Hong Ding (Dept of Clinical Pharmacology, Covance Laboratories Inc., Madison, WI) for SAS statistical analysis, and Nehad M. Alajez (Ontario Cancer Institute, Toronto, Ontario, Canada) for discussion and communication throughout the experiments. This work was supported by NIH CA 56103 grant to OJF and Woeber Fund for Breast Cancer Research to XC.

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Authors and Affiliations

  1. Department of Immunology, University of Pittsburgh School of Medicine, 200 Lothrop Street, Pittsburgh, PA, 15261, USA

    Xiaochuan Chen & Olivera J. Finn

  2. Garden State Cancer Center, Center for Molecular Medicine and Immunology, 520 Belleville Ave, Belleville, NJ, 07109, USA

    Xiaochuan Chen

  3. Department of Pathology, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15261, USA

    Wentao Gao

  4. Center for Biotechnology and Institute of Molecular Medicine, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15261, USA

    Andrea Gambotto

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  1. Xiaochuan Chen
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  2. Wentao Gao
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  4. Olivera J. Finn
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Correspondence to Olivera J. Finn.

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Chen, X., Gao, W., Gambotto, A. et al. Lentiviral vectors encoding human MUC1-specific, MHC-unrestricted single-chain TCR and a fusion suicide gene: potential for universal and safe cancer immunotherapy. Cancer Immunol Immunother 58, 977–987 (2009). https://doi.org/10.1007/s00262-008-0624-0

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