Distinct Signaling of Coreceptors Regulates Specific Metabolism Pathways and Impacts Memory Development in CAR T Cells

Chimeric antigen receptors (CARs) redirect T cell cytotoxicity against cancer cells, providing a promising approach to cancer immunotherapy. Despite extensive clinical use, the attributes of CAR co-stimulatory domains that impact persistence and resistance to exhaustion of CAR-T cells remain largely undefined. Here, we report the influence of signaling domains of coreceptors CD28 and 4-1BB on the metabolic characteristics of human CAR T cells. Inclusion of 4-1BB in the CAR architecture promoted the outgrowth of CD8+ central memory T cells that had significantly enhanced respiratory capacity, increased fatty acid oxidation and enhanced mitochondrial biogenesis. In contrast, CAR T cells with CD28 domains yielded effector memory cells with a genetic signature consistent with enhanced glycolysis. These results provide, at least in part, a mechanistic insight into the differential persistence of CAR-T cells expressing 4-1BB or CD28 signaling domains in clinical trials and inform the design of future CAR T cell therapies. [Display omitted] •CAR T cells containing 4-1BB signaling domains have enhanced in vitro persistence•Increased central memory differentiation of CD8+ CAR T cells expressing 4-1BB•CD28-enforced metabolic reprograming enhances glycolytic metabolism•Mitochondrial biogenesis is selectively induced in 4-1BB CAR T cells Despite the successes of chimeric antigen receptor (CAR) T cells in treating hematologic malignancies, the impact of various CAR signaling domains remains unclear. Kawalekar et al. show that the choice of signaling domain can metabolically reprogram T cells to alter its mitochondrial biogenesis and persistence.