Tuning charge density of chimeric antigen receptor optimizes tonic signaling and CAR-T cell fitness  

作　　者：Jian Chen Shizhen Qiu Wentao Li Kun Wang Yu Zhang Han Yang Baichuan Liu Guangfei Li Li Li Min Chen Junjie Lan Jiahua Niu Peijie He Lei Cheng Gaofeng Fan Xin Liu Xianmin Song Chenqi Xu Haitao Wu Haopeng Wang 

机构地区：[1]School of Life Science and Technology,ShanghaiTech University,Shanghai,China [2]ENT institute and Department of Otorhinolaryngology,Eye&ENT Hospital,Fudan University,Shanghai,China [3]State Key Laboratory of Molecular Biology,Shanghai Institute of Biochemistry and Cell Biology,Center for Excellence in Molecular Cell Science,Chinese Academy of Sciences,University of Chinese Academy of Sciences,Shanghai,China [4]Xiamen Shuangshi Middle School of Fujian,Xiamen,Fujian,China [5]Department of Hematology,Shanghai General Hospital,Shanghai Jiaotong University School of Medicine,Shanghai,China [6]School of Life Science,Hangzhou Institute for Advanced Study,University of Chinese Academy of Sciences,Hangzhou,Zhejiang,China [7]Shanghai Clinical Research and Trial Center,Shanghai,China [8]Present address:CeMM,Research Center for Molecular Medicine of the Austrian Academy of Sciences,Vienna,Austria

出　　处：《Cell Research》2023年第5期341-354,共14页细胞研究（英文版）

基　　金：the Animal Facility at the National Facility for Protein Science in Shanghai,Zhangjiang Lab for providing the support in mouse housing and care,and Molecular and Cell Biology Core Facility(MCBCF)and Molecular Imaging Core Facility(MICF)at the School of Life Science and Technology,ShanghaiTech University for providing technical support.This work was supported by grants from the National Key R&D Program of China(2019YFA0111000 to H.Wang);the Double First-Class Initiative Fund of ShanghaiTech University,Shanghai Frontiers Science Center for Biomacromolecules and Precision Medicine at ShanghaiTech University,and Shanghai Local College Capacity Building Project(22010502700);J.C.was granted by Youth Foundation of the National Natural Science Foundation of China(82102863);Original Exploration Project of Shanghai Natural Science Foundation(21ZR1480200);H.Wu was granted by the National Natural Science Foundation of China(82271149)and Shanghai Technology Innovation Project(21Y11912000);C.X.was granted by CAS Young Scientists in Basic Research Program(YSBR-014);CAS Shanghai Branch Special Plan for Basic Research(JCYJ-SHFY-2022-009)。

摘　　要：Tonic signaling of chimeric antigen receptor(CAR),i.e.,the spontaneous CAR activation in the absence of tumor antigen stimulation,is considered to be a pivotal event controlling CAR-T efficacy.However,the molecular mechanism underlying the spontaneous CAR signals remains elusive.Here,we unveil that positively charged patches(PCPs)on the surface of the CAR antigenbinding domain mediate CAR clustering and result in CAR tonic signaling.For CARs with high tonic signaling(e.g.,GD2.CAR and CSPG4.CAR),reducing PCPs on CARs or boosting ionic strength in the culture medium during ex vivo CAR-T cell expansion minimizes spontaneous CAR activation and alleviates CAR-T cell exhaustion.In contrast,introducing PCPs into the CAR with weak tonic signaling,such as CD19.CAR,results in improved in vivo persistence and superior antitumor function.These results demonstrate that CAR tonic signaling is induced and maintained by PCP-mediated CAR clustering.Notably,the mutations we generated to alter the PCPs maintain the antigen-binding affinity and specificity of the CAR.Therefore,our findings suggest that the rational tuning of PCPs to optimize tonic signaling and in vivo fitness of CAR-T cells is a promising design strategy for the nextgeneration CAR.

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分 类 号：R730[医药卫生—肿瘤]