Department of Basic Medical, Bengbu Medical College, Bengbu, AH, China.
Department of Life Sciences, Bengbu Medical College, Bengbu, AH, China.
Bioengineered. 2022 Apr;13(4):9216-9232. doi: 10.1080/21655979.2022.2058149.
Non-small cell lung cancer (NSCLC) is considered to be one of the most prevalent and fatal malignancies, with a poor survival rate. Chimeric antigen receptor T cell (CAR-T) cell therapy is one of the most exciting directions in the field of Cellular immunotherapy. Therefore, CAR-T cells that target c-Met have been developed for use in NSCLC therapy and might be a potential therapeutic strategy. The anti c-Met scFv structure was fused with the transmembrane and intracellular domains. Using a lentiviral vector to load the c-Met CAR gene, then transfected the c-Met CAR lentiviral into human T cells to obtain the second generation c-Met CAR-T expressing CARs stably. In vitro co-culture, experiments revealed that CAR-T cells have high proliferative activity and the potential to secrete cytokines (IL-2, TNF-α, and IFN-γ). c-Met CAR-T cells showed special cellular cytotoxicity in LDH release assay. A subcutaneous tumor model in nude mice was used to test the anticancer effectiveness of c-met CAR-T cells in vivo. For c-Met positive NSCLC tissue, according to tumor volume, weight, fluorescence intensity, and immunohistochemical detection, c-Met CAR-T cells had stronger tumor growth suppression compared to untransduced T cells. HE staining revealed that c-Met CAR-T cells did not produced side effects in nude mice. Taken together, we provided useful method to generate c-Met CAR- T cells, which exhibit enhanced cytotoxicity against NSCLC cells in vitro and in vivo. Thus, providing a new therapeutic avenue for treating NSCLC clinically. (1) c-Met CAR-T capable of stably expressing c-Met CARs were constructed.(2) c-Met CAR-T have strong anti-tumor ability and proliferation ability in vitro.(3) c-Met CAR-T can effectively inhibit the growth of A549 cells subcutaneous xenografts.
非小细胞肺癌(NSCLC)被认为是最常见和最致命的恶性肿瘤之一,其生存率较低。嵌合抗原受体 T 细胞(CAR-T)细胞疗法是细胞免疫疗法领域最令人兴奋的方向之一。因此,已经开发出针对 c-Met 的 CAR-T 细胞用于 NSCLC 治疗,可能是一种潜在的治疗策略。抗 c-Met scFv 结构与跨膜和细胞内结构域融合。使用慢病毒载体装载 c-Met CAR 基因,然后将 c-Met CAR 慢病毒转染到人 T 细胞中,获得稳定表达 CAR 的第二代 c-Met CAR-T。体外共培养实验表明,CAR-T 细胞具有高增殖活性和分泌细胞因子(IL-2、TNF-α 和 IFN-γ)的潜力。c-Met CAR-T 细胞在 LDH 释放实验中表现出特殊的细胞毒性。在裸鼠皮下肿瘤模型中测试 c-met CAR-T 细胞在体内的抗癌效果。对于 c-Met 阳性 NSCLC 组织,根据肿瘤体积、重量、荧光强度和免疫组织化学检测,c-Met CAR-T 细胞比未转导的 T 细胞具有更强的肿瘤生长抑制作用。HE 染色显示 c-Met CAR-T 细胞在裸鼠中没有产生副作用。总之,我们提供了一种有用的方法来产生 c-Met CAR-T 细胞,这些细胞在体外和体内对 NSCLC 细胞表现出增强的细胞毒性。因此,为临床治疗 NSCLC 提供了新的治疗途径。(1) 构建了能够稳定表达 c-Met CAR 的 c-Met CAR-T。(2) c-Met CAR-T 在体外具有强大的抗肿瘤能力和增殖能力。(3) c-Met CAR-T 能有效抑制 A549 细胞皮下移植瘤的生长。
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