Jiang Wei, Li Tao, Guo Jiaojiao, Wang Jingjing, Jia Lizhou, Shi Xiao, Yang Tingting, Jiao Ruonan, Wei Xin, Feng Zhenqing, Tang Qi, Ji Guozhong
Department of Gastroenterology, The Second Affiliated Hospital of Nanjing Medical University, Nanjing, China.
Jiangsu Key Lab of Cancer Biomarkers, Prevention and Treatment, Collaborative Innovation Center for Cancer Personalized Medicine, Nanjing Medical University, Nanjing, China.
Front Oncol. 2021 Mar 10;11:546586. doi: 10.3389/fonc.2021.546586. eCollection 2021.
T cells expressing chimeric antigen receptors, especially CD19 CAR-T cells have exhibited effective antitumor activities in B cell malignancies, but due to several factors such as antigen escape effects and tumor microenvironment, their curative potential in hepatocellular carcinoma has not been encouraging. To reduce the antigen escape risk of hepatocellular carcinoma, this study was to design and construct a bispecific CAR targeting c-Met and PD-L1. c-Met/PD-L1 CAR-T cells were obtained by lentiviral transfection, and the transfection efficiency was monitored by flow cytometry analysis. LDH release assays were used to elucidate the efficacy of c-Met/PD-L1 CAR-T cells on hepatocellular carcinoma cells. In addition, xenograft models bearing human hepatocellular carcinoma were constructed to detect the antitumor effect of c-Met/PD-L1 CAR-T cells . The results shown that this bispecific CAR was manufactured successfully, T cells modified with this bispecific CAR demonstrated improved antitumor activities against c-Met and PD-L1 positive hepatocellular carcinoma cells when compared with those of monovalent c-Met CAR-T cells or PD-L1 CAR-T cells but shown no distinct cytotoxicity on hepatocytes . experiments shown that c-Met/PD-L1 CAR-T cells significantly inhibited tumor growth and improve survival persistence compared with other groups. These results suggested that the design of single-chain, bi-specific c-Met/PD-L1 CAR-T is more effective than that of monovalent c-Met CAR-T for the treatment of hepatocellular carcinoma., and this bi-specific c-Met/PD-L1 CAR is rational and implementable with current T-cell engineering technology.
表达嵌合抗原受体的T细胞，尤其是CD19嵌合抗原受体T细胞（CAR-T细胞），在B细胞恶性肿瘤中已表现出有效的抗肿瘤活性，但由于抗原逃逸效应和肿瘤微环境等多种因素，它们在肝细胞癌中的治疗潜力并不乐观。为降低肝细胞癌的抗原逃逸风险，本研究旨在设计并构建一种靶向c-Met和PD-L1的双特异性嵌合抗原受体。通过慢病毒转染获得c-Met/PD-L1 CAR-T细胞，并通过流式细胞术分析监测转染效率。采用乳酸脱氢酶释放试验来阐明c-Met/PD-L1 CAR-T细胞对肝癌细胞的疗效。此外，构建了携带人肝细胞癌的异种移植模型，以检测c-Met/PD-L1 CAR-T细胞的抗肿瘤作用。结果表明，这种双特异性嵌合抗原受体成功制备，与单价c-Met CAR-T细胞或PD-L1 CAR-T细胞相比，用这种双特异性嵌合抗原受体修饰的T细胞对c-Met和PD-L1阳性肝癌细胞表现出更强的抗肿瘤活性，但对肝细胞无明显细胞毒性。实验表明，与其他组相比，c-Met/PD-L1 CAR-T细胞显著抑制肿瘤生长并提高生存持久性。这些结果表明，单链双特异性c-Met/PD-L1 CAR-T的设计在治疗肝细胞癌方面比单价c-Met CAR-T更有效，并且这种双特异性c-Met/PD-L1嵌合抗原受体在当前T细胞工程技术下是合理且可行的。
