Engagement of an optimized lentiviral vector enhances the expression and cytotoxicity of CAR in human NK cells.
Engagement of an optimized lentiviral vector enhances the expression and cytotoxicity of CAR in human NK cells.
复制标题
DOI：
10.1016/j.molimm.2023.01.010
复制
发表时间：
2023-02
影响因子：
3.6
通讯作者：
Changjiang Guo;Han Chen;Jie Yu;Hui Lu;Qing Xia;Xiaojuan Li;Xiali Guo;Tongrong Wang;Lingtong Zhi;Zhiyuan Niu;Wuling Zhu
中科院分区：
文献类型：
--
作者：
Changjiang Guo;Han Chen;Jie Yu;Hui Lu;Qing Xia;Xiaojuan Li;Xiali Guo;Tongrong Wang;Lingtong Zhi;Zhiyuan Niu;Wuling Zhu
免疫嵌合抗原受体（CAR）修饰的T或NK细胞（CAR-T/NK）已经成为一种新的疾病治疗形式。慢病毒载体（LV）通常用于工程化NK细胞以有效表达汽车。本研究报道了单启动子和双启动子LV对工程化NK细胞CAR表达和细胞毒性的影响。我们构建了第三代基于NKG 2D的CAR，通过靶向多达8种应激诱导配体（NKG 2DLs）来杀死癌细胞。我们的研究结果表明，通过利用优化的单启动子pCDHsp而不是原始的双启动子pCDHdp，CAR在HEK-293 T或NK 92细胞中表现出更高的表达水平和与GFP报告基因的更高的共表达一致性。在嘌呤霉素选择后，pCDHsp产生稳健的CAR表达和增强的工程化NK细胞的体外细胞毒性。因此，推荐用单启动子pCDHsp慢病毒载体感染以制备CAR工程化NK细胞。该研究有助于优化CAR-NK细胞的生产并增强其功能活性，为CAR-NK细胞产品提供更好，更均匀的质量。
Adoptive chimeric antigen receptor (CAR)-modified T or NK cells (CAR-T/NK) have emerged as a novel form of disease treatment. Lentiviral vectors (LVs) are commonly employed to engineer NK cells for the efficient expression of CARs. This study reported the influence of single-promoter and dual-promoter LVs on the CAR expression and cytotoxicity of engineered NK cells. We constructed a third-generation NKG2D-based CAR that kills cancer cells by targeting up to eight stress-induced ligands (NKG2DLs). Our results demonstrated that the CAR exhibits both a higher expression level and a higher coexpression concordance with the GFP reporter in HEK-293T or NK92 cells by utilizing the optimized single-promoter pCDHsp rather than the original dual-promoter pCDHdp. After puromycin selection, the pCDHsp produces robust CAR expression and enhanced in vitro cytotoxicity of engineered NK cells. Therefore, infection with a single-promoter pCDHsp lentivector is recommended to prepare CAR-engineered NK cells. This research helps to optimize the production of CAR-NK cells and enhance their functional activity, to provide CAR-NK cell products with better and more uniform quality.