Zhang Pengchao, Feng Xuejia, Niu Xiangyun, Liu Zhongming, Li Minghui, Liu Maoxuan, Yan Dehong, Zhang Guizhong, Wan Xiaochun
Center for Protein and Cell-based Drugs, Institute of Biomedicine and Biotechnology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, 518055, People's Republic of China.
University of Chinese Academy of Sciences, Beijing, 100049, PR China.
Exp Hematol Oncol. 2025 Mar 3;14(1):28. doi: 10.1186/s40164-025-00618-7.
Chimeric antigen receptor (CAR)-NK therapy holds great potential for tumor treatment, but current CAR designs are primarily optimized for T cells, raising concerns about their suitability for NK cells. This study compared two dominant CAR designs used in T cells-CD28-CD3ζ (28z) and 4-1BB-CD3ζ (BBz)-and found that CD28 costimulation offers superior functionality in NK cells. 28z CAR-NK cells exhibited significantly better activation, cytotoxicity, and in vivo anti-tumor efficacy than BBz CAR-NK cells, with similar persistence and tumor infiltration. 28z CAR more effectively recruited the ZAP70 kinase and upregulated multiple key factors involved in immune activation, potentially augmenting CAR-NK cell function. MAP3K8, a kinase involved in inflammation and the MAPK signaling pathway, was identified as a critical mediator in enhancing 28z CAR-NK cell function. Silencing or inhibiting MAP3K8 impaired the anti-tumor activity of 28z CAR-NK cells, while its overexpression substantially improved the function of BBz CAR-NK cells. These findings provide new insights into how CD28 costimulation boosts CAR-NK cell efficacy, supporting its use into NK cell-specific CARs for cancer immunotherapy, and highlight MAP3K8 as a potential target for optimizing BBz CAR-NK cell therapy.
嵌合抗原受体（CAR）-NK疗法在肿瘤治疗方面具有巨大潜力，但目前的CAR设计主要是针对T细胞进行优化的，这引发了对其是否适用于NK细胞的担忧。本研究比较了两种在T细胞中使用的主要CAR设计——CD28-CD3ζ（28z）和4-1BB-CD3ζ（BBz），发现CD28共刺激在NK细胞中具有更优越的功能。与BBz CAR-NK细胞相比，28z CAR-NK细胞表现出显著更好的激活、细胞毒性和体内抗肿瘤疗效，且具有相似的持久性和肿瘤浸润能力。28z CAR更有效地招募ZAP70激酶并上调多个参与免疫激活的关键因子，可能增强CAR-NK细胞功能。MAP3K8是一种参与炎症和MAPK信号通路的激酶，被确定为增强28z CAR-NK细胞功能的关键介质。沉默或抑制MAP3K8会损害28z CAR-NK细胞的抗肿瘤活性，而其过表达则显著改善了BBz CAR-NK细胞的功能。这些发现为CD28共刺激如何提高CAR-NK细胞疗效提供了新的见解，支持将其用于癌症免疫治疗的NK细胞特异性CAR，并突出了MAP3K8作为优化BBz CAR-NK细胞疗法的潜在靶点。
