CRISPR-mediated TCR replacement generates superior anticancer transgenic T-cells

Legut, Mateusz, Dolton, Garry, Mian, Afsar, Ottmann, Oliver ORCID: https://orcid.org/0000-0001-9559-1330 and Sewell, Andrew ORCID: https://orcid.org/0000-0003-3194-3135 2018. CRISPR-mediated TCR replacement generates superior anticancer transgenic T-cells. Blood 131 (3) , pp. 311-322. 10.1182/blood-2017-05-787598

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Abstract

Adoptive transfer of T-cells genetically modified to express a cancer-specific T-cell receptor (TCR) has shown significant therapeutic potential for both hematological and solid tumors. However, a major issue of transducing T-cells with a transgenic TCR is the pre-existing expression of TCRs in the recipient cells. These endogenous TCRs compete with the transgenic TCR for surface expression and allow mixed dimer formation. Mixed dimers, formed by mispairing between the endogenous and transgenic TCRs, may harbor autoreactive specificities. To circumvent these problems, we designed a system where the endogenous TCR-β is knocked out from the recipient cells using CRISPR/Cas9 technology, simultaneously with transduction with a cancer-reactive receptor of choice. This TCR replacement strategy resulted in markedly increased surface expression of transgenic αβ and γδ TCRs, which in turn translated to a stronger, and more polyfunctional, response of engineered T-cells to their target cancer cell lines. Additionally, the TCR+CRISPR modified T-cells were up to a thousandfold more sensitive to antigen than standard TCR-transduced T-cells or conventional model proxy systems used for studying TCR activity. Finally, transduction with a pan-cancer reactive γδ TCR used in conjunction with CRISPR/Cas9 knockout of the endogenous αβ TCR resulted in more efficient redirection of CD4+ and CD8+ T-cells against a panel of established blood cancers and primary, patient-derived B acute lymphoblastic leukemia blasts compared to standard TCR transfer. Our results suggest that TCR transfer combined with genome editing could lead to new improved generations of cancer immunotherapies.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Medicine
Subjects:	R Medicine > R Medicine (General)
Publisher:	American Society of Hematology
ISSN:	0006-4971
Funders:	Wellcome Trust
Date of First Compliant Deposit:	30 November 2017
Date of Acceptance:	4 November 2017
Last Modified:	15 Nov 2023 16:26
URI:	https://orca.cardiff.ac.uk/id/eprint/106690

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