TCF1^high progenitor CD8⁺ T cells mediate the efficacy of immunotherapy; however, the mechanisms that govern their generation and maintenance are poorly understood. Here, we show that targeting glycolysis through deletion of pyruvate kinase muscle 2 (PKM2) results in elevated pentose phosphate pathway (PPP) activity, leading to enrichment of a TCF1^high progenitor-exhausted-like phenotype and increased responsiveness to PD-1 blockade in vivo. PKM2^KO CD8⁺ T cells showed reduced glycolytic flux, accumulation of glycolytic intermediates and PPP metabolites and increased PPP cycling as determined by 1,2-¹³C glucose carbon tracing. Small molecule agonism of the PPP without acute glycolytic impairment skewed CD8⁺ T cells toward a TCF1^high population, generated a unique transcriptional landscape and adoptive transfer of agonist-treated CD8⁺ T cells enhanced tumor control in mice in combination with PD-1 blockade and promoted tumor killing in patient-derived tumor organoids. Our study demonstrates a new metabolic reprogramming that contributes to a progenitor-like T cell state promoting immunotherapy efficacy.

TCF1^高祖细胞 CD8⁺ T 细胞介导免疫治疗的疗效;然而，人们对控制其生成和维护的机制知之甚少。在这里，我们表明通过删除丙酮酸激酶肌肉 2 （PKM2） 靶向糖酵解导致磷酸戊糖途径 （PPP） 活性升高，导致 TCF1^高祖细胞耗竭样表型的富集和体内对 PD-1 阻断的反应性增加。PKM2^KO CD8⁺ T 细胞显示糖酵解通量降低，糖酵解中间体和 PPP 代谢物积累，PPP 循环增加，通过 1,2-13 C 葡萄糖碳示踪确定。没有急性糖酵解损伤的 PPP 的小分子激动作用使 CD8 ⁺ T 细胞偏向 TCF1^高群体，产生了独特的转录景观和激动剂处理的 CD8 ⁺ T 细胞的过继转移，与 PD-1 阻断联合增强了小鼠的肿瘤控制，并促进了患者来源的肿瘤类器官中的肿瘤杀伤。我们的研究证明了一种新的代谢重编程，它有助于祖细胞样 T 细胞状态促进免疫治疗效果。