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Multi-cistronic vector encoding optimized safety switch for adoptive therapy with T-cell receptor-modified T cells

Gene Therapy volume 20, pages 861–867 (2013)Cite this article

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Abstract

T-cell receptor (TCR) gene transfer is an attractive strategy to equip T cells with defined antigen-specific TCRs using short-term in vitro procedures to target both hematological malignancies and solid tumors. TCR gene transfer poses different safety issues that might warrant the inclusion of a suicide gene. High affinity TCRs may result in on-target toxicity, and off-target reactivity directed against healthy tissue can be observed due to mixed TCR dimers. Inclusion of a suicide gene as a safety switch may abrogate these unwanted toxicities. Human CD20 has been proposed as a nonimmunogenic suicide gene targeted by widely used clinical-grade anti-CD20 antibodies that can additionally function as a selection marker. However, transduction of T cells with a multi-cistronic vector encoding both TCR and CD20 resulted in poor coexpression. In this study, we demonstrated that codon optimization of TCR and CD20 resulted in profound coexpression of both the preferentially expressed antigen in melanoma (PRAME)-TCR and CD20, allowing selective as well as efficient elimination of these engineered T cells in vitro. These results demonstrate the great potential of codon optimized CD20 to be broadly used in clinical trials as a safety switch.

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Acknowledgements

This work was supported by the Dutch Cancer Society (grant NKB 2007–3927) and ZonMw (grant ZonMw 433.00.001). We thank Michel Kester, Dirk van der Steen, Guido de Roo and Menno van der Hoorn for expert technical assistance.

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Authors and Affiliations

  1. Department of Hematology, Leiden University Medical Center, Leiden, The Netherlands

    M M van Loenen, R de Boer, R S Hagedoorn, V Jankipersadsing, A L Amir, J H F Falkenburg & M H M Heemskerk

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  1. M M van Loenen
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  2. R de Boer
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  3. R S Hagedoorn
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  4. V Jankipersadsing
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  5. A L Amir
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  6. J H F Falkenburg
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  7. M H M Heemskerk
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Correspondence to M M van Loenen.

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van Loenen, M., de Boer, R., Hagedoorn, R. et al. Multi-cistronic vector encoding optimized safety switch for adoptive therapy with T-cell receptor-modified T cells. Gene Ther 20, 861–867 (2013). https://doi.org/10.1038/gt.2013.4

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