KPNA2-Mediated MYC Nuclear Import Enhances CAR-T Cell Memory and Potentiates Antitumor Efficacy

Chimeric antigen receptor (CAR)-T cell therapy demonstrates remarkable clinical efficacy, yet limited persistence and functional exhaustion impede durable responses. While memory-like phenotypes are associated with sustained CAR-T function and improved clinical outcomes, the underlying molecular mechanisms of CAR-T cell memory maintenance remain incompletely defined and we still lack actionable strategy to sufficiently promote CAR-T cell memory formation. Here, we identified that Karyopherin Subunit Alpha 2 (KPNA2)-mediated MYC nuclear import robustly enhanced CAR-T cell memory and antitumor function. In clinical CAR-T cell products, KPNA2 expression and MYC pathway activation are correlated with superior therapeutic potency. Overexpressing MYC showed negligible effect on CAR-T cell function due to insufficient nuclear import; in contrast, incorporating KPNA2 in CAR-T cells facilitated MYC nuclear import, augmented memory formation, enhanced cytotoxicity and antitumor activity both in vitro and in vivo. Such functional improvement was not associated with risks of transformation in KPNA2-overexpressing CAR-T cells. Mechanistically, our integrated transcriptomic and epigenomic analyses revealed that KPNA2 increased MYC occupancy at memory-associated gene loci, upregulating T cell memory programs. Furthermore, virtual drug screening identified the small molecule TMS which potentiates KPNA2-MYC interaction. TMS treatment of CAR-T cells enhanced MYC nuclear accumulation, promoted memory-oriented transcriptional profiles, and improved antitumor potency in preclinical models. These findings established KPNA2-MYC interaction as a pivotal mechanism governing CAR-T cell memory formation, providing genetic and pharmacological strategies to advance therapeutic T cell design and manufacturing.