Integrated drug profiling and CRISPR screening identify essential pathways for CAR T-cell cytotoxicity

Olli Dufva, Jan Koski, Pilvi Maliniemi, Aleksandr Ianevski, Jay Klievink, Judith Leitner, Petri Pölönen, Helena Hohtari, Khalid Saeed, Tiina Hannunen, Pekka Ellonen, Peter Steinberger, Matti Kankainen, Tero Aittokallio, Mikko A.I. Keränen, Matti Korhonen, Satu Mustjoki*

*Corresponding author for this work

Research output: Contribution to journalArticleScientificpeer-review

53 Citations (Scopus)
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Abstract

Chimeric antigen receptor (CAR) T-cell therapy has proven effective in relapsed and refractory B-cell malignancies, but resistance and relapses still occur. Better understanding of mechanisms influencing CAR T-cell cytotoxicity and the potential for modulation using small-molecule drugs could improve current immunotherapies. Here, we systematically investigated druggable mechanisms of CAR T-cell cytotoxicity using >500 small-molecule drugs and genome-scale CRISPR-Cas9 loss-of-function screens. We identified several tyrosine kinase inhibitors that inhibit CAR T-cell cytotoxicity by impairing T-cell signaling transcriptional activity. In contrast, the apoptotic modulator drugs SMAC mimetics sensitized B-cell acute lymphoblastic leukemia and diffuse large B-cell lymphoma cells to anti-CD19 CAR T cells. CRISPR screens identified death receptor signaling through FADD and TNFRSF10B (TRAIL-R2) as a key mediator of CAR T-cell cytotoxicity and elucidated the RIPK1-dependent mechanism of sensitization by SMAC mimetics. Death receptor expression varied across genetic subtypes of B-cell malignancies, suggesting a link between mechanisms of CAR T-cell cytotoxicity and cancer genetics. These results implicate death receptor signaling as an important mediator of cancer cell sensitivity to CAR T-cell cytotoxicity, with potential for pharmacological targeting to enhance cancer immunotherapy. The screening data provide a resource of immunomodulatory properties of cancer drugs and genetic mechanisms influencing CAR T-cell cytotoxicity. Key Points: • Survey of immunomodulatory effects of >500 drugs identifies SMAC mimetics as sensitizers to CAR T-cell cytotoxicity. • Genome-scale CRISPR screen reveals essentiality of death receptor signaling for CAR T-cell cytotoxicity.

Original languageEnglish
Pages (from-to)597-609
Number of pages13
JournalBLOOD
Volume135
Issue number9
DOIs
Publication statusPublished - 27 Feb 2020
MoE publication typeA1 Journal article-refereed

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