The adenylyl cyclase activator forskolin pretreatment reprograms CAR-T cells toward a stem-like state to enhance solid tumor therapy.

Chimeric antigen receptor (CAR)-T cell therapy has achieved remarkable success in hematological malignancies; however, its efficacy against solid tumors remains limited, primarily due to T cell exhaustion. Enhancing memory-like properties and stemness of T cells has emerged as a promising strategy to overcome this barrier. In this study, we demonstrate that Forskolin, a stem cell-inducing compound, reprograms CAR-T cells toward a memory-like state characterized by elevated stemness. Forskolin-pretreated CAR-T cells exhibited enriched memory phenotypes, reduced exhaustion, and enhanced anti-tumor activity both in vitro and in vivo. Transcriptomic profiling further revealed upregulation of CXCR4 and BCL6 following Forskolin treatment, and functional blockade of these pathways attenuated the beneficial effects of Forskolin, underscoring their critical roles in reinforcing CAR-T cell stemness. Our findings highlight a novel strategy that integrates stem cell-reprogramming agents to enhance CAR-T cell stemness and provide potential therapeutic targets to improve the efficacy of CAR-T cell therapy against solid tumors.