A GMP-compliant protocol to expand and transfect cancer patient T cells with mRNA encoding a tumor-specific chimeric antigen receptor

Krug C, Wiesinger M, Abken H, Schuler-Thurner B, Schuler G, Dörrie J, Schaft N (2014)

Publication Type: Journal article

Publication year: 2014

Journal

Cancer Immunology Immunotherapy Springer Verlag (Germany)

Publisher: Springer Verlag (Germany)

Book Volume: 63

Pages Range: 999-1008

Journal Issue: 10

DOI: 10.1007/s00262-014-1572-5

Abstract

Chimeric antigen receptors (CARs), which combine an antibody-derived binding domain (single chain fragment variable) with T-cell-activating signaling domains, have become a promising tool in the adoptive cellular therapy of cancer. Retro- and lenti-viral transductions are currently the standard methods to equip T cells with a CAR; permanent CAR expression, however, harbors several risks like uncontrolled auto-reactivity. Modification of T cells by electroporation with CAR-encoding RNA to achieve transient expression likely circumvents these difficulties. We here present a GMP-compliant protocol to activate and expand T cells for clinical application. The protocol is optimized in particular to produce CAR-modified T cells in clinically sufficient numbers under full GMP-compliance from late-stage cancer patients. This protocol allows the generation of 6.7 × 10(8) CAR-expressing T cells from one patient leukapheresis. The CAR-engineered T cells produced pro-inflammatory cytokines after stimulation with antigen-bearing tumor cells and lysed tumor cells in an antigen-specific manner. This functional capacity was maintained after cryopreservation. Taken together, we provide a clinically applicable protocol to transiently engineer sufficient numbers of antigen-specific patient T cells for use in adoptive cell therapy of cancer.

Authors with CRIS profile

Christian Krug Lehrstuhl für Genetik Beatrice Schuler-Thurner Lehrstuhl für Haut- und Geschlechtskrankheiten Gerold Schuler Lehrstuhl für Haut- und Geschlechtskrankheiten Jan Dörrie Department of Dermatology Niels Schaft Department of Dermatology

Involved external institutions

Universität zu Köln DE Germany (DE)

How to cite

APA:

Krug, C., Wiesinger, M., Abken, H., Schuler-Thurner, B., Schuler, G., Dörrie, J., & Schaft, N. (2014). A GMP-compliant protocol to expand and transfect cancer patient T cells with mRNA encoding a tumor-specific chimeric antigen receptor. Cancer Immunology Immunotherapy, 63(10), 999-1008. https://doi.org/10.1007/s00262-014-1572-5

MLA:

Krug, Christian, et al. "A GMP-compliant protocol to expand and transfect cancer patient T cells with mRNA encoding a tumor-specific chimeric antigen receptor." Cancer Immunology Immunotherapy 63.10 (2014): 999-1008.

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