美国北卡罗来纳大学Gianpietro Dotti、Yang Xu团队的最新研究，利用多激酶抑制剂筛选发现了保护类嵌合抗原受体T细胞（CAR T细胞）干细胞样的抑制剂。2025年1月8日出版的《自然—免疫学》发表了这项成果。

具有T_SCM样表型特征的嵌合抗原受体T细胞，可促进持续抗肿瘤效应。研究人员对以激酶为重点的化合物集进行了无偏倚、自动化高通量筛选，以确定能维持人类T干细胞（T_SCM）样CAR T细胞的激酶抑制剂 (KIs)。研究发现了三种KI：UNC10225387B、UNC10225263A和UNC10112761A，这三种KI在体外联合使用，可增加健康供体和癌症患者的CD45RA⁺CCR7⁺TCF1^hi T_SCM细胞样CAR T细胞的频率。

经KI处理的CAR T细胞在体外和体内小鼠肿瘤模型中，均显示出更强的抗肿瘤效果。KI鸡尾酒能优先维持初始T细胞来源CAR T细胞的T_SCM细胞样表型，并导致转录组变化，而不会阻止T细胞活化或改变染色质组织。KI鸡尾酒以剂量依赖性方式靶向的特定激酶ITK、ADCK3、MAP3K4和CDK13，这与T_SCM细胞样CAR T细胞的保护直接相关。

单独或联合敲除这些激酶可富集T_SCM细胞样CAR T细胞，但只有在KI鸡尾酒存在的情况下产生的CAR T细胞，才会在肿瘤细胞刺激下表现出强大的增殖和分化能力。总之，对目标激酶的短暂药理抑制，可保持CAR T细胞的干细胞样特征，并提高其抗肿瘤活性。

附：英文原文

Title: A multi-kinase inhibitor screen identifies inhibitors preserving stem-cell-like chimeric antigen receptor T cells

Author: 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

Issue&Volume: 2025-01-08

Abstract: Chimeric antigen receptor T cells (CAR T cells) with T stem (TSCM) 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 TSCM cell-like CAR T cells. We identified three KIs, UNC10225387B, UNC10225263A and UNC10112761A, that combined in vitro increased the frequency of CD45RA+CCR7+TCF1hi TSCM 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 TSCM 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 TSCM cell-like CAR T cells. Knockdown of these kinases individually or in combination enriches for TSCM 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.

DOI: 10.1038/s41590-024-02042-1

Source: https://www.nature.com/articles/s41590-024-02042-1