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[Preprint]. 2023 Jan 27:2023.01.26.525725. [Version 1] doi: 10.1101/2023.01.26.525725

Costimulatory domains direct distinct fates of CAR-driven T cell dysfunction

Mehmet Emrah Selli, Jack H Landmann, Marina Terekhova, John Lattin, Amanda Heard, Yu-Sung Hsu, Tien-Ching Chang, Jufang Chang, John Warrington, Helen Ha, Natalie Kingston, Graham Hogg, Michael Slade, Melissa M Berrien-Elliot, Mark Foster, Samantha Kersting-Schadek, Agata Gruszczynska, David DeNardo, Todd A Fehniger, Maxim Artyomov, Nathan Singh
PMCID: PMC9901009  PMID: 36747791

Abstract

Chimeric antigen receptor (CAR) engineered T cells often fail to enact effector functions after infusion into patients. Understanding the biological pathways that lead CAR T cells to failure is of critical importance in the design of more effective therapies. We developed and validated an in vitro model that drives T cell dysfunction through chronic CAR activation and interrogated how CAR costimulatory domains contribute to T cell failure. We found that dysfunctional CD28-based CARs targeting CD19 bear hallmarks of classical T cell exhaustion while dysfunctional 41BB-based CARs are phenotypically, transcriptionally and epigenetically distinct. We confirmed activation of this unique transcriptional program in CAR T cells that failed to control clinical disease. Further, we demonstrate that 41BB-dependent activation of the transcription factor FOXO3 is a significant contributor to this dysfunction and disruption of FOXO3 improves CAR T cell function. These findings identify that chronic activation of 41BB leads to novel state of T cell dysfunction that can be alleviated by genetic modification of FOXO3.

Summary

Chronic stimulation of CARs containing the 41BB costimulatory domain leads to a novel state of T cell dysfunction that is distinct from T cell exhaustion.

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