Enhanced Effector Activity of Mediator Kinase Module Deficient CAR-T Cells

• DOI: 10.1101/2022.09.02.506235
• Publication Date: 2022-09-05T03:50:14Z
• Authors: Katherine A. Freitas; Julia A. Belk; Elena Sotillo; Bence Daniel; Katalin Sandor; Dorota Klysz; Vandon T. Duong; Kylie Burdsall; Peng Xu; Meena Malipatlolla; Micah G. Donovan; Evan W. Weber; Robbie G. Majzner; Howard Y. Chang; Joaquin M. Espinosa; Ansuman T. Satpathy; Crystal L. Mackall
• Keywords: 0301 basic medicine; 03 medical and health sciences

Abstract Adoptive T cell immune therapies mediate impressive clinical benefit in a fraction of patients, but anti-tumor effects are often limited by inadequate potency. To identify genes limiting effector function, we conducted genome-wide CRISPR knock-out screens human primary CAR-T cells. The top hits were MED12 and CCNC , components the cyclin-dependent kinase (CDK) module Mediator complex, an evolutionarily conserved regulator gene transcription. or deficient cells manifest increased expansion, cytokine production, metabolic fitness, activity reduced terminal differentiation. Chemical inhibition CDK8/19 recapitulated some features genetic loss including expansion. showed widespread selective increases chromatin accessibility, MED1 occupancy, H3K27 acetylation at enhancers used transcription factors playing critical role fate, several STAT AP1 family members. most pronounced enhancement was observed for STAT5 which manifested as sensitivity to IL-2 These results link induced transcriptional coactivation with programming CDK target enhancing potency responses. One Sentence Summary is differentiation, small molecule-based this enhances