Si Wen, Fan Ying-Ying, Qiu Shi-Zhen, Li Xin, Wu Er-Yi, Ju Jian-Qi, Huang Wen, Wang Hao-Peng, Wei Ping
Center for Cell and Gene Circuit Design, CAS Key Laboratory of Quantitative Engineering Biology, Shenzhen Institute of Synthetic Biology, Shenzhen Institute of Advanced Technology, Chinese Academy of Sciences, Shenzhen 518055, China.
Center for Quantitative Biology and Peking-Tsinghua Joint Center for Life Sciences, Academy for Advanced Interdisciplinary Studies, Peking University, Beijing 100871, China.
iScience. 2023 Mar 30;26(4):106529. doi: 10.1016/j.isci.2023.106529. eCollection 2023 Apr 21.
Chimeric antigen receptor (CAR)-T cells have shown great promise in cancer therapy. However, the anti-tumor efficiency is limited due to the CAR-induced T cell apoptosis or exhaustion. The intracellular domain of CAR comprised of various signaling modules orchestrates CAR-T cell behaviors. The modularity of CAR signaling domain functions as the "mainboard" to assemble diversified downstream signaling components. Here, we implemented the modular recombination strategy to construct a library of CARs with synthetic co-signaling modules adopted from immunoglobin-like superfamily (IgSF) and tumor necrosis factor receptor superfamily (TNFRSF). We quantitatively characterized the signaling behaviors of these recombinants by both NFAT and NF-κB reporter, and identified a set of new CARs with diverse signaling behaviors. Specifically, the 28(NM)-BB(MC) CAR-T cells exhibited improved cytotoxicity and T cell persistence. The synthetic approach can promote our understanding of the signaling principles of CAR molecule, and provide a powerful tool box for CAR-T cell engineering.
嵌合抗原受体（CAR）-T细胞在癌症治疗中显示出巨大的前景。然而，由于CAR诱导的T细胞凋亡或耗竭，其抗肿瘤效率受到限制。CAR的细胞内结构域由各种信号模块组成，协调CAR-T细胞的行为。CAR信号结构域的模块化充当“主板”，用于组装多样化的下游信号组件。在这里，我们实施了模块化重组策略，构建了一个CAR文库，其中采用了来自免疫球蛋白样超家族（IgSF）和肿瘤坏死因子受体超家族（TNFRSF）的合成共刺激信号模块。我们通过NFAT和NF-κB报告基因定量表征了这些重组体的信号行为，并鉴定出一组具有不同信号行为的新型CAR。具体而言，28（NM）-BB（MC）CAR-T细胞表现出改善的细胞毒性和T细胞持久性。这种合成方法可以促进我们对CAR分子信号原理的理解，并为CAR-T细胞工程提供一个强大的工具箱。
