Ren Quan, Zu Yingling, Su Hongchang, Lu Qiumei, Xiang Bin, Luo Yanping, Zhang Jishuai, Song Yongping
Department of Hematology, The Affiliated Cancer Hospital of Zhengzhou University and Henan Cancer Hospital, Zhengzhou, Henan, 450008, China.
Shenzhen Pregene Biopharma Company Ltd, Shenzhen, 518118, China.
Exp Hematol Oncol. 2023 Nov 27;12(1):98. doi: 10.1186/s40164-023-00461-8.
Natural killer (NK) cells are promising alternatives for the production of "off-the-shelf" CAR products, posing a lower risk of cytokine release syndrome (CRS) than CAR-T cells. We synthesized four single VHH-directed anti-BCMA CARs, incorporating various intracellular regions (2B4 versus CD28) and hinge domains (CD28 versus IgG1) and ectopically producing IL-15. NK cells derived from peripheral blood (PB) were expanded ex vivo by K562-mbIL21 feeder cells. Stable CAR transduction was obtained through lentiviral transduction with the BaEV-Rless pseudotyped lentiviral vector. BCMA-CD28-IL15 CAR-NK cells with ectopic expression of IL-15 exhibited superior cytotoxicity were compared to BCMA-CD28 CAR-NK cells lacking IL-15 and BCMA-hIgG1-IL15 CAR-NK cells with an IgG1 hinge domain. We further assessed the cytotoxic capabilities of BCMA-2B4-IL15 CAR-NK cells with 2B4 intracellular domain. The BCMA-CD28-IL15 CAR-NK cells revealed stronger cytotoxicity and higher cytokine secretion against BCMA tumor cells than BCMA-2B4-IL15 CAR-NK cells in vitro. In the MM.1S-Luc mouse model, BCMA-CD28-IL15 CAR-NK inhibited the growth of tumor cells and prolonged mouse survival. These results show that the single VHH-directed BCMA CAR-NK cells exhibited remarkable specific killing ability, making them a potential candidate for immunotherapy in multiple myeloma treatment.
自然杀伤（NK）细胞是生产“现成可用”嵌合抗原受体（CAR）产品的有前景的替代方案，与CAR-T细胞相比，其细胞因子释放综合征（CRS）风险更低。我们合成了四种单VHH导向的抗BCMA CAR，整合了各种细胞内区域（2B4与CD28）、铰链区（CD28与IgG1）并异位表达白细胞介素-15（IL-15）。来自外周血（PB）的NK细胞通过K562-mbIL21饲养细胞在体外进行扩增。通过使用BaEV-Rless假型慢病毒载体进行慢病毒转导获得稳定的CAR转导。将异位表达IL-15且具有卓越细胞毒性的BCMA-CD28-IL15 CAR-NK细胞与缺乏IL-15的BCMA-CD28 CAR-NK细胞以及具有IgG1铰链区的BCMA-hIgG1-IL15 CAR-NK细胞进行比较。我们进一步评估了具有2B4细胞内结构域的BCMA-2B4-IL15 CAR-NK细胞的细胞毒性能力。在体外，BCMA-CD28-IL15 CAR-NK细胞对BCMA肿瘤细胞显示出比BCMA-2B4-IL15 CAR-NK细胞更强的细胞毒性和更高的细胞因子分泌。在MM.1S-Luc小鼠模型中，BCMA-CD28-IL15 CAR-NK抑制了肿瘤细胞的生长并延长了小鼠存活时间。这些结果表明，单VHH导向的BCMA CAR-NK细胞表现出显著的特异性杀伤能力，使其成为多发性骨髓瘤治疗中免疫疗法的潜在候选者。
