Increased NFAT activity with dual CAR stimulation in CD19xCD22 CAR T-cells is associated with decreased exhaustion and improved survival

作者：Alexander W. Rankin, Catherine Pham‐Danis, Amanda J Novak, Etienne Danis, Terry J. Fry, M. Eric Kohler · 发表于：Journal for ImmunoTherapy of Cancer · 年份：2025 · DOI：10.1136/jitc-2025-011971 · 被引用次数：3 · 研究领域：CAR-T cell therapy research、T-cell and B-cell Immunology、Cutaneous lymphoproliferative disorders research

BACKGROUND: Chimeric antigen receptor (CAR) T-cell therapy is effective in treating B-cell malignancies, however relapse due to lack of CAR persistence and antigen-modulated escape remains common. Multiple strategies to simultaneously target CD19 and CD22 have been able to reduce antigen-modulated escape but not completely eliminate relapse. A bicistronic CAR construct consisting of a CD19 CAR incorporating the CD28 costimulatory domain paired with a CD22 CAR incorporating a 4-1BB costimulatory domain (CD19xCD22) demonstrated superior preclinical activity compared with other configurations and is currently under clinical investigation (NCT05098613, NCT05442515, NCT06559189). We hypothesized that simultaneous activation of CD28-containing and 4-1BB-containing CAR molecules not only allows for targeting of both antigens but creates a unique signal which enhances CAR T-cell function and efficacy. METHODS: We tested CD19xCD22 CAR T-cells generated from primary human T-cells against NALM6 with wild-type expression of CD19 and CD22 (CD19+/CD22+) or CRISPR/Cas9 knockout of one or both antigens (CD19+/CD22-, CD19-/CD22+, CD19-/CD22-) to interrogate the effect of dual-CAR stimulation on T-cell function, signaling, and in vivo efficacy in xenograft models. RESULTS: In vitro proliferation and cytokine production of CD19xCD22 CAR T-cells were primarily driven by activation of the CD19-28z CAR, however the CD22-BBz CAR drove equivalent cytotoxicity. Dual-CAR stimulation of CD19xCD22 CAR T...