LAUSR

Abstract Chimeric antigen receptor (CAR) T cells are potent agents for recognizing and eliminating tumors, and have achieved remarkable success in the treatment of patients with refractory leukemia and lymphoma. However, dysfunction of T cells, including exhaustion, is an inevitable obstacle for persistent curative effects. Here, the authors initially found that calcium signaling is hyperactivated via sustained tonic signaling in CAR‐T cells. Next, it is revealed that the store‐operated calcium entry (SOCE) inhibitor BTP‐2, but not the calcium chelator BAPTA‐AM, markedly diminishes CAR‐T cell exhaustion and terminal differentiation of CAR‐T cells in both tonic signaling and tumor antigen exposure models. Furthermore, BTP‐2 pretreated CAR‐T cells show improved antitumor potency and prolonged survival in vivo. Mechanistically, transcriptome and metabolite analyses reveal that treatment with BTP‐2 significantly downregulate SOCE‐calcineurin‐nuclear factor of activated T‐cells (NFAT) and glycolysis pathways. Together, the results indicate that modulating the SOCE‐calcineurin‐NFAT pathway in CAR‐T cells renders them resistant to exhaustion, thereby yielding CAR products with enhanced antitumor potency.