Arrangement of VL and VH domains in VRC01-Based chimeric antigen receptor (CAR) affects functions and exhaustion status of CAR-T cells
Arrangement of VL and VH domains in VRC01-Based chimeric antigen receptor (CAR) affects functions and exhaustion status of CAR-T cells
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基于VRC01的嵌合抗原受体（CAR）中VL和VH结构域的排列影响CAR-T细胞的功能和耗竭状态
DOI：
10.1089/regen.2021.0005
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发表时间：
2021
期刊：
影响因子：
--
通讯作者：
Su Lishan
中科院分区：
文献类型：
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作者：
Nunoya Jun-ichi;Teramura Eijiro;Masuda Michiaki;Su Lishan
背景：使用表达嵌合抗原受体(CAR)的工程T细胞进行免疫治疗的病例越来越多。CAR分子有一个抗原结合域，通常由抗体衍生的VHland VHdomain组成。目的：研究Vland VH域在CAR中的相对排列是否会影响CAR-T细胞的耗竭状态，尤其是携带疱疹病毒进入介体(Hvem)衍生的共刺激信号域(CSSD)。方法：我们构建了不同排列方式的携带hvem来源的CSSD和HIV-1广泛中和抗体VRC01的vland VHA域的CARS作为模型系统，结果：在转导的T细胞中，VL-VHCAR的表达水平高于VH-VLCAR，具有更强的效应功能。表型分析表明，VH-VLCAR-T细胞包含两个群体：一个具有较高的CAR表达和CAR介导的紧张性信号与更多的耗竭表型相关，另一个具有相反的特征。有趣的是，尽管VL-VHCAR-T细胞表达更高水平的CAR，但CAR介导的紧张性信号较低，与疲惫人群的比例较低相关，这导致在同源抗原模拟中有较高的扩增。这些结果表明，尽管CAR高表达，但VL-VHCAR-T细胞似乎逃脱了CAR介导的强直信号诱导的耗竭。结论：VRC01-CAR中VLAND VHH域的排列不仅对CAR-T细胞的功能有重要影响，而且对CAR-T细胞的耗竭状态也有重要影响。因此，临床前验证Vland VHdomain与不同CSSD在CAR中的相对位置有助于开发有效的CAR-T细胞介导的免疫治疗。
Background:Immunotherapy using engineered T cells expressing chimeric antigen receptor (CAR) is used in an increasing number of cases. The CAR molecule has an antigen-binding domain usually consisting of an antibody-derived VLand VHdomains. The arrangement of VLand VHdomains has been shown to affect CAR characteristics and functions of the CAR-transduced T (CAR-T) cells.Objectives:This study aims to clarify whether the relative arrangement of VLand VHdomains in CAR affects the exhaustion status of CAR-T cells especially bearing herpes virus entry mediator (HVEM)-derived co-stimulatory signal domain (CSSD).Method:We generated CARs bearing the HVEM-derived CSSD and the VLand VHdomains derived from HIV-1 broadly neutralizing antibody VRC01 in different arrangements (VL-VHor VH-VL) as a model system, and compared effector functions and characteristics of the CAR-T cells.Results:In the transduced T cells, the VL-VHCAR was expressed at higher levels and led to more potent effector functions than the VH-VLCAR. Phenotypic analysis showed that the VH-VLCAR-T cells contained two populations: one with higher CAR expression and CAR-mediated tonic signaling associated with more exhausted phenotype, and the other with the opposite characteristics. Interestingly, the VL-VHCAR-T cells, although expressing higher levels of CAR, exhibited lower CAR-mediated tonic signaling associated with lower percentage of exhausted population, which led to higher expansion upon cognate antigen simulation. These results suggested that the VL-VHCAR-T cells appeared to escape exhaustion induced by CAR-mediated tonic signaling despite the high CAR expression.Conclusions:The arrangement of VLand VHdomains in VRC01-based CAR appears to play important roles in not only functions but also exhaustion status of CAR-T cells with HVEM-derived CSSD. Therefore, preclinical validation of the relative position of VLand VHdomains combined with various CSSDs in CAR could assist in developing effective CAR-T cell-mediated immunotherapy.