Song Feifei, Tsahouridis Ourania, Stucchi Simone, Walhart Tara, Mendell Sophie, Hardy P Brian, Axtman Matthew, Guduru Shiva K R, Gilbert Thomas S K, Graves Lee M, Herring Laura E, Savoldo Barbara, Ma Xingcong, Woodcock Mark, Milner Justin J, Ivanova Anastasia, Pearce Kenneth H, Xu Yang, Dotti Gianpietro
Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
Center for Integrative Chemical Biology and Drug Discovery, Eshelman School of Pharmacy, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA.
Nat Immunol. 2025 Feb;26(2):279-293. doi: 10.1038/s41590-024-02042-1. Epub 2025 Jan 8.
Chimeric antigen receptor T cells (CAR T cells) with T stem (T) cell-like phenotypic characteristics promote sustained antitumor effects. We performed an unbiased and automated high-throughput screen of a kinase-focused compound set to identify kinase inhibitors (KIs) that preserve human T cell-like CAR T cells. We identified three KIs, UNC10225387B, UNC10225263A and UNC10112761A, that combined in vitro increased the frequency of CD45RACCR7TCF1 T cell-like CAR T cells from both healthy donors and patients with cancer. KI-treated CAR T cells showed enhanced antitumor effects both in vitro and in vivo in mouse tumor models. The KI cocktail maintains T cell-like phenotype preferentially in CAR T cells originating from naive T cells and causes transcriptomic changes without arresting T cell activation or modulating the chromatin organization. Specific kinases, ITK, ADCK3, MAP3K4 and CDK13, targeted by the KI cocktail in a dose-dependent manner are directly associated with the preservation of T cell-like CAR T cells. Knockdown of these kinases individually or in combination enriches for T cell-like CAR T cells, but only CAR T cells generated in the presence of the KI cocktail show robust expansion and differentiation on stimulation with tumor cells. Overall, transient pharmacological inhibition of strategically targeted kinases maintains stem-like features in CAR T cells and improves their antitumor activity.
具有T干细胞样表型特征的嵌合抗原受体T细胞（CAR T细胞）可促进持续的抗肿瘤作用。我们对一组以激酶为重点的化合物进行了无偏倚的自动化高通量筛选，以鉴定能保留人T细胞样CAR T细胞的激酶抑制剂（KIs）。我们鉴定出三种激酶抑制剂，UNC10225387B、UNC10225263A和UNC10112761A，它们在体外联合使用可增加来自健康供体和癌症患者的CD45RA⁺CCR7⁺TCF1⁺ T细胞样CAR T细胞的频率。在小鼠肿瘤模型中，经激酶抑制剂处理的CAR T细胞在体外和体内均显示出增强的抗肿瘤作用。激酶抑制剂鸡尾酒优先在源自初始T细胞的CAR T细胞中维持T细胞样表型，并引起转录组变化，而不会阻止T细胞活化或调节染色质组织。激酶抑制剂鸡尾酒以剂量依赖性方式靶向的特定激酶ITK、ADCK3、MAP3K4和CDK13与T细胞样CAR T细胞的保留直接相关。单独或联合敲低这些激酶可富集T细胞样CAR T细胞，但只有在存在激酶抑制剂鸡尾酒的情况下产生的CAR T细胞在受到肿瘤细胞刺激时才显示出强劲的扩增和分化。总体而言，对策略性靶向激酶的短暂药理抑制可维持CAR T细胞中的干细胞样特征并提高其抗肿瘤活性。
