Wang Xindi, Luo Wenjing, Chen Zhaozhao, Li Chenggong, Zhou Jie, Huang Zhongpei, Tang Lu, Wu Jianghua, Wu Zhuolin, Li Yingying, Zhang Yinqiang, Kang Yun, Liu Qiaolin, Xu Jia, Xiong Wei, Deng Jun, Mei Heng, Hu Yu
Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China.
Hubei Clinical Medical Center of Cell Therapy for Neoplastic Disease, Wuhan, Hubei, China.
J Immunother Cancer. 2025 Jun 15;13(6):e010822. doi: 10.1136/jitc-2024-010822.
Chimeric antigen receptor (CAR)-natural killer (NK) cell therapy has demonstrated safety and feasibility in clinical settings; however, limited efficacy due to intrinsic dysfunction and extrinsic suppression remains an unresolved issue. T cells provide multifaceted support to NK cell-mediated responses. Here, we aimed to design a novel CD19-targeted CAR-NK, engineered with secreted interleukin-15 and C-C motif chemokine ligand 21 (ie, 15×21 CAR-NK), capable of recruiting and cooperating with T cells.
We characterized 15×21 CAR-NK cells by performing experiments in vitro and in mouse models, and conducting RNA sequencing.
15×21 CAR-NK cells exhibit strong capabilities in cytotoxicity, cytokine production, effector molecule expression, and T-cell recruitment in vitro. Cooperation with T cells promoted efficient tumor-cell elimination, alleviated mutual exhaustion phenotypes, and enhanced the expression of effector molecules/receptors. The recruitment and cooperative effects also result in effective tumor control in mouse models. In addition, 15×21 CAR-NK cells strongly enrich the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway - a key intracellular signaling cascade that is associated with enhanced downstream pro-survival signaling, anti-apoptotic ability, mitochondrial function, and cytotoxicity.
Our study highlights the intrinsic advantages and extrinsic T-cell cooperative benefits of 15×21 CAR-NK cells, providing a promising strategy for NK-cell-based immunotherapy.
嵌合抗原受体（CAR）-自然杀伤（NK）细胞疗法已在临床环境中证明了安全性和可行性；然而，由于内在功能障碍和外在抑制导致的疗效有限仍是一个未解决的问题。T细胞为NK细胞介导的反应提供多方面的支持。在此，我们旨在设计一种新型的靶向CD19的CAR-NK，其工程化表达分泌型白细胞介素-15和C-C基序趋化因子配体21（即15×21 CAR-NK），能够招募T细胞并与之合作。
我们通过体外实验、小鼠模型实验以及RNA测序对15×21 CAR-NK细胞进行了表征。
15×21 CAR-NK细胞在体外具有强大的细胞毒性、细胞因子产生能力、效应分子表达能力和T细胞招募能力。与T细胞合作促进了肿瘤细胞的有效清除，减轻了相互耗竭的表型，并增强了效应分子/受体的表达。这种招募和合作效应在小鼠模型中也实现了有效的肿瘤控制。此外，15×21 CAR-NK细胞强烈富集磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（AKT）/雷帕霉素哺乳动物靶蛋白（mTOR）信号通路——这是一个关键的细胞内信号级联反应，与增强下游促生存信号、抗凋亡能力、线粒体功能和细胞毒性相关。
我们的研究突出了15×21 CAR-NK细胞的内在优势和外在T细胞合作益处，为基于NK细胞的免疫治疗提供了一种有前景的策略。
