Relapses of CD19-expressing leukemia in patients who achieved initial remission after CART cell treatment have been reported to correlate with poor CART cells persistence. Sustained tonic signaling or strong activation drives CART cell differentiation and exhaustion, which limit the therapeutic efficacy and persistence of CART cells. Here, we identified dasatinib as the optimal candidate to prevent or reverse both CD28/CART and 4-1BB/CART cell differentiation and exhaustion during ex vivo expansion, which profoundly enhanced the therapeutic efficacy and in vivo persistence. Moreover, strong activation-induced CART cells differentiation, exhaustion and apoptosis driven by CD3/CD28 stimulation or antigen exposure were dramatically prevented or reversed by dasatinib treatment. Mechanistically, dasatinib markedly reduced the phosphorylation of Src and Lck, and downregulated the expression of genes involved in CAR signaling pathways, which resulted in the optimization of cell differentiation, exhaustion and apoptosis-related gene expression. Our study proposes a promising pharmacological approach for optimizing CART cells manufacture, and provides an experimental basis for reinvigorating CART cells in clinical application.

中文翻译：

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达沙替尼通过抑制细胞分化和衰竭增强嵌合抗原受体 T 细胞的抗白血病功效

据报道，在 CART 细胞治疗后获得初始缓解的患者中表达 CD19 的白血病复发与 CART 细胞持久性差相关。持续的强直信号或强激活驱动 CART 细胞分化和衰竭，这限制了 CART 细胞的治疗效果和持久性。在这里，我们确定达沙替尼是在体外扩增过程中预防或逆转 CD28/CART 和 4-1BB/CART 细胞分化和耗竭的最佳候选药物，这极大地提高了治疗效果和体内持久性。此外，达沙替尼治疗显着阻止或逆转了由 CD3/CD28 刺激或抗原暴露驱动的强烈激活诱导的 CART 细胞分化、衰竭和凋亡。从机制上讲，达沙替尼显着降低了 Src 和 Lck 的磷酸化，并下调了参与 CAR 信号通路的基因的表达，从而优化了细胞分化、衰竭和凋亡相关基因的表达。我们的研究提出了一种优化 CART 细胞制造的有前景的药理学方法，并为在临床应用中重振 CART 细胞提供了实验基础。