新型双串联CD19/BCMA嵌合抗原受体T细胞的特性研究，有望用于治疗多发性骨髓瘤。
Characterization of novel dual tandem CD19/BCMA chimeric antigen receptor T cells to potentially treat multiple myeloma.
作者信息
Kang Liqing, Zhang Jian, Li Minghao, Xu Nan, Qi Wei, Tan Jingwen, Lou Xiaoyan, Yu Zhou, Sun Juanjuan, Wang Zhenkun, Fu Chengcheng, Tang Xiaowen, Dai Haiping, Chen Jia, Wu Depei, Yu Lei
机构信息
1Institute of Biomedical Engineering and Technology, Shanghai Engineering Research Center of Molecular Therapeutics and New Drug Development, School of Chemistry and Molecular Engineering, East China Normal University, NO, 3663 North Zhongshan Road, Shanghai, 200065 China.
2National Clinical Research Center for Hematologic Diseases, Jiangsu Institute of Hematology, The First Affiliated Hospital of Soochow University, Suzhou, China.
出版信息
Biomark Res. 2020 May 13;8:14. doi: 10.1186/s40364-020-00192-6. eCollection 2020.
BACKGROUND
Treatment with chimeric antigen receptor (CAR)-engineered T cells directed against the B-cell maturation antigen (BCMA) promoted transient recovery from multiple myeloma (MM). However, the absence of this antigen on immature plasma cells may limit the efficacy of this modality and facilitate relapse. The purpose of this study is to characterize a novel CAR that includes both a single-chain variable fragment (scFv)-BCMA and an scFv-CD19 in tandem orientation (tan-CAR) in an attempt to target both BCMA and CD19 expression on MM cells.
METHOD
The scFv sequences from the anti-CD19 antibody FMC63 and the anti-BCMA antibody C11D5.3 were ligated in tandem with transmembrane and T-cell signaling domains to generate the tan-CAR construct. Specificity and efficacy of activated tan-CAR T cells were analyzed using in vitro proliferation, cytokine release, and cytolysis assays. We also evaluated the in vivo efficacy with a xenograft mouse model that included target tumor cells that expressed CD19 or BCMA and compared the results to those obtained with conventional CAR T cells.
RESULTS
The in vitro studies revealed specific activation of tan-CAR T cells by K562 cells that overexpressed CD19 and/or BCMA. Cell proliferation, cytokine release, and cytolytic activity were all comparable to the responses of single scFv CAR T cells. Importantly, in vivo studies of tan-CAR T cells revealed specific inhibition of tumor growth in the mouse xenograft model that included cells expressing both CD19 and BCMA. Systemic administration of tan-CAR T cells resulted in complete tumor remission, in contrast to the reduced efficacies of BCMA-CAR T and CD19-CAR T alone in this setting.
CONCLUSION
We report the successful design and execution of novel tan-CAR T cells that promote significant anti-tumor efficacy against both CD19 and BCMA antigen-positive tumor cells in vitro and in vivo. The data from this study reveal a novel strategy that may help to reduce the rate of relapse in the treatment with single scFv-CAR T cells.
背景
用靶向B细胞成熟抗原（BCMA）的嵌合抗原受体（CAR）工程化T细胞进行治疗可促进多发性骨髓瘤（MM）的短暂缓解。然而，未成熟浆细胞上缺乏这种抗原可能会限制这种治疗方式的疗效并促进复发。本研究的目的是表征一种新型CAR，其包含串联排列的单链可变片段（scFv）-BCMA和scFv-CD19（tan-CAR），试图靶向MM细胞上的BCMA和CD19表达。
方法
将抗CD19抗体FMC63和抗BCMA抗体C11D5.3的scFv序列与跨膜和T细胞信号结构域串联连接，以生成tan-CAR构建体。使用体外增殖、细胞因子释放和细胞溶解试验分析活化的tan-CAR T细胞的特异性和功效。我们还使用异种移植小鼠模型评估了体内功效，该模型包括表达CD19或BCMA的靶肿瘤细胞，并将结果与传统CAR T细胞获得的结果进行比较。
结果
体外研究显示，过表达CD19和/或BCMA的K562细胞可特异性激活tan-CAR T细胞。细胞增殖、细胞因子释放和细胞溶解活性均与单scFv CAR T细胞的反应相当。重要的是，tan-CAR T细胞的体内研究显示，在包含表达CD19和BCMA的细胞的小鼠异种移植模型中，肿瘤生长受到特异性抑制。与单独使用BCMA-CAR T和CD19-CAR T在此情况下疗效降低形成对比的是，全身给予tan-CAR T细胞可导致肿瘤完全缓解。
结论
我们报告了新型tan-CAR T细胞的成功设计和实施，其在体外和体内均对CD19和BCMA抗原阳性肿瘤细胞具有显著的抗肿瘤功效。本研究的数据揭示了一种新策略，可能有助于降低单scFv-CAR T细胞治疗中的复发率。
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