ZAP327 signaling domain–driven chimeric antigen receptor generates robust and long-term antitumor immunity in mouse models

Chimeric antigen receptor (CAR) T-cell therapy has shown impressive clinical responses in the treatment of blood cancers, but high percentages of disease relapse one year after T cell infusion and severe toxicities associated with CAR-T cell therapy remain major issues. Here, we report the construction of CARs with a ZAP-70-derived signaling domain (ZAP327) that enhances therapeutic antitumor activity with increasing in vivo T cell persistence. Importantly, ZAP327-driven CAR-T cells markedly reduced cytokine release and expression of T cell exhaustion markers. Mechanistically, we show that the ZAP327 domain tuned down TCR signaling, increased the pools of stem-like memory T cells, and exhibited metabolic features associated with memory T cells by utilizing the oxidative phosphorylation pathway. These results highlight the therapeutic potential of ZAP327-driven CAR-T cells to overcome the limitations of the current CAR-T cell therapies and enhance the potency and persistence of antitumor T cell responses in solid tumors as well.