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  • Original Article - Enabling Technologies
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Original Article – Enabling Technologies

Antithymidylate resistance enables transgene selection and cell survival for T cells in the presence of 5-fluorouracil and antifolates

Gene Therapy volume 23, pages 119–128 (2016)Cite this article

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Abstract

Antithymidylates (AThy) constitute a class of drugs used in the treatment of cancers such as lung, colon, breast and pancreas. These drugs inhibit DNA synthesis by targeting the enzymes dihydrofolate reductase (DHFR) and/or thymidylate synthase (TYMS). AThys effectively inhibit cancer cells, and also inhibit T cells, preventing anticancer immunity, which might otherwise develop from AThy-induced cancer destruction. We establish that T cells expressing mutant DHFR—DHFR L22F, F31S (DHFRFS)—and/or mutant TYMS—TYMS T51S, G52S (TYMSSS)—effectively survive in toxic concentrations of AThys methotrexate, pemetrexed and 5-fluorouracil. Furthermore, we show that DHFRFS permitted rapid selection of an inducible suicide transgene in T cells. These findings demonstrate that AThy resistances prevent AThy cytotoxicity to T cells while permitting selection of important transgenes. This technological development could enhance in vitro and in vivo survival and selection of T-cell therapeutics being designed for a broad range of cancers.

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Acknowledgements

We gratefully acknowledge the dedicated work of Helen Huls, Dr Natalya Belousova, Dr Harjeet Singh, Dr Pappanaicken Kumar and Tiejuan Mi for maintaining a functional laboratory environment. We also acknowledge Dr Dean Lee, Dr Dat Tran and Dr George McNamara for critical appraisal of the manuscript and constructive suggestions in manuscript preparation. The MD Anderson South Campus Flow Cytometry Facility, NCI Cancer Center Support Grant P30CA16672, kindly assisted in data acquisition for some experiments.

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Authors and Affiliations

  1. Division of Pediatrics, Children’s Cancer Hospital, The University of Texas MD Anderson Cancer Center, Unit 907, Houston, TX, USA

    D Rushworth, S Olivares & L J N Cooper

  2. The University of Texas Graduate School of Biomedical Sciences at Houston, Houston, TX, USA

    D Rushworth, A Alpert, R Santana-Carrero & L J N Cooper

  3. University of Puerto Rico School of Medicine, San Juan, Puerto Rico, United States Minor Outlying Islands

    R Santana-Carrero

  4. Bellicum Pharmaceuticals, Houston, TX, USA

    D Spencer

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  1. D Rushworth
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Correspondence to L J N Cooper.

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Competing interests

Dr Cooper founded and owns InCellerate Inc. He has patents with Sangamo BioSciences on artificial nucleases. He consults with Targazyme Inc. (formerly American Stem Cells Inc.), GE Healthcare, Ferring Pharmaceuticals Inc. and Bristol-Myers Squibb. Dr Cooper receives honoraria from Miltenyi Biotec. He has agreed to license technologies developed in his laboratory to Intrexon Corp. and Ziopharma Oncology Inc. He has numerous patents in the field of immunotherapy and genetic engineering.

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Rushworth, D., Alpert, A., Santana-Carrero, R. et al. Antithymidylate resistance enables transgene selection and cell survival for T cells in the presence of 5-fluorouracil and antifolates. Gene Ther 23, 119–128 (2016). https://doi.org/10.1038/gt.2015.88

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  • DOI: https://doi.org/10.1038/gt.2015.88

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