Zhong M J, Xu Y X, Xing H Y, Tang K J, Tian Z, Rao Q, Wang M, Wang J X
Institute of Hematology & Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College; State Key Laboratory of Experimental Hematology; National Clinical Research Center for Blood Diseases, Tianjin 300020, China.
Zhonghua Xue Ye Xue Za Zhi. 2019 Oct 14;40(10):804-811. doi: 10.3760/cma.j.issn.0253-2727.2019.10.002.
To construct the BCMA-CAR using the B-cell maturation antigen (BCMA) specific ligand APRIL as antigen binding region and to validate the effect of BCMA-CAR modified T cells (BCMA-CAR-T) on myeloma cells. The BCMA-CAR was constructed using the BCMA specific ligand APRIL as antigen binding domain and 4-1BB as the costimulatory domain. The specific cytotoxicity against BCMA(+) myeloma cell lines and primary multiple myeloma (MM) cells in vitro were evaluated. In addition, BCMA(+) myeloma xenograft mouse model was established to assess the anti-tumor effect of BCMA-CAR-T cell therapy in vivo. BCMA-CAR-T cells could specifically kill BCMA(+) myeloma cell lines (For BCMA-CAR-T cells, BCMA(+) cells are almost undetectable in the E∶T ratio of 1∶4) and MM patients' bone marrow mononuclear cells (the proportion of residual cells in BCMA-CAR-T and vector-T groups was 16.0% 66.85%, =0.003) with significant degranulation (CAR-T and vector-T cells cocultured with MM1.S, H929 and U266 had degranulation levels of 33.30% 5.62%, 16.97% 2.95% and 25.87% 2.97%, respectively, <0.001) and cytokines release (<0.01) in vitro. In a human BCMA(+) myeloma xenograft mouse model, BCMA-CAR-T cells could significantly prolong the survival of mice (The median survival time of mice treated with BCMA-CAR-T and vector-T cells was 87.5 days and 67.5 days, respectively, <0.001) . The ligand-based BCMA-CAR-T cells could be a promising strategy for BCMA(+) multiple myeloma treatment.
构建以B细胞成熟抗原（BCMA）特异性配体增殖诱导配体（APRIL）作为抗原结合区的BCMA嵌合抗原受体（BCMA-CAR），并验证BCMA-CAR修饰的T细胞（BCMA-CAR-T）对骨髓瘤细胞的作用。以BCMA特异性配体APRIL作为抗原结合结构域、4-1BB作为共刺激结构域构建BCMA-CAR。评估其对BCMA(+)骨髓瘤细胞系和原发性多发性骨髓瘤（MM）细胞的体外特异性细胞毒性。此外，建立BCMA(+)骨髓瘤异种移植小鼠模型，以评估BCMA-CAR-T细胞疗法在体内的抗肿瘤作用。BCMA-CAR-T细胞能够特异性杀伤BCMA(+)骨髓瘤细胞系（对于BCMA-CAR-T细胞，在效靶比为1∶4时，BCMA(+)细胞几乎检测不到）和MM患者的骨髓单个核细胞（BCMA-CAR-T组和载体-T组残余细胞比例分别为16.0%和66.85%，P=0.003），体外具有显著的脱颗粒现象（与MM1.S、H929和U266共培养的CAR-T和载体-T细胞脱颗粒水平分别为33.30%±5.62%、16.97%±2.95%和25.87%±2.97%，P<0.001）和细胞因子释放（P<0.01）。在人BCMA(+)骨髓瘤异种移植小鼠模型中，BCMA-CAR-T细胞能够显著延长小鼠生存期（接受BCMA-CAR-T和载体-T细胞治疗的小鼠中位生存时间分别为87.5天和67.5天，P<0.001）。基于配体的BCMA-CAR-T细胞可能是治疗BCMA(+)多发性骨髓瘤的一种有前景的策略。
