肿瘤细胞治疗研究
英文原题：Preclinical assessment of CAR-NK cell-mediated killing efficacy and pharmacokinetics in a rapid zebrafish xenograft model of metastatic breast cancer.
Preclinical assessment of CAR-NK cell-mediated killing efficacy and pharmacokinetics in a rapid zebrafish xenograft model of metastatic breast cancer.
我们的数据提示，斑马鱼幼体可用于快速且经济高效地体内评估 CAR-NK 细胞效力，并预测患者对治疗的反应。
自然杀伤（NK）细胞是癌症过继免疫治疗中颇具吸引力的效应细胞。采用嵌合抗原受体（CAR）改造的原代NK细胞和NK-92细胞开展的首批人体研究，在疗效和安全性方面结果令人鼓舞。为进一步改进治疗策略，并以个体化方式检验CAR-NK细胞疗效，需要利用患者来源的肿瘤样本开展临床前筛选。斑马鱼（Danio rerio）胚胎和幼体具有出色的活细胞成像特性，是研究患者来源肿瘤细胞生长和播散的有吸引力的异种移植模型。将细胞注入其循环系统，可用于研究转移、癌细胞与免疫细胞相互作用以及肿瘤细胞对抗癌药物的反应。本研究建立了斑马鱼幼体异种移植模型：先注入转移性乳腺癌细胞，再将CAR-NK细胞注入库维耶导管（DoC），以在体内评估CAR-NK细胞对转移性乳腺癌的疗效。我们使用两种分别靶向PD-L1和ErbB2的CAR-NK细胞系验证该系统功能，即PD-L1.CAR NK-92和ErbB2.CAR NK-92细胞，靶向表达PD-L1的MDA-MB-231细胞系和表达ErbB2的MDA-MB-453细胞系。注入的癌细胞具有活性，并分布于幼体外周区域，包括尾部造血组织（CHT），模拟癌细胞归巢至造血部位。2.5小时后注入的CAR-NK细胞迁移至CHT，并迅速清除整个机体内的单个癌细胞。未改造的NK-92细胞在体内也表现出轻微细胞毒性。共聚焦活细胞成像显示，CAR-NK细胞在血管内迁移并与MDA-MB-231细胞实时相互作用，解释了其快速而有效的体内细胞毒作用。因此，我们的数据提示，斑马鱼幼体可用于快速、低成本地在体评估CAR-NK细胞效力，并预测患者对治疗的反应。
Natural killer (NK) cells are attractive effectors for adoptive immunotherapy of cancer. Results from first-in-human studies using chimeric antigen receptor (CAR)-engineered primary NK cells and NK-92 cells are encouraging in terms of efficacy and safety. In order to further improve treatment strategies and to test the efficacy of CAR-NK cells in a personalized manner, preclinical screening assays using patient-derived tumor samples are needed. Zebrafish ( Danio rerio ) embryos and larvae represent an attractive xenograft model to study growth and dissemination of patient-derived tumor cells because of their superb live cell imaging properties. Injection into the organism's circulation allows investigation of metastasis, cancer cell-to-immune cell-interactions and studies of the tumor cell response to anti-cancer drugs. Here, we established a zebrafish larval xenograft model to test the efficacy of CAR-NK cells against metastatic breast cancer in vivo by injecting metastatic breast cancer cells followed by CAR-NK cell injection into the Duct of Cuvier (DoC). We validated the functionality of the system with two different CAR-NK cell lines specific for PD-L1 and ErbB2 (PD-L1.CAR NK-92 and ErbB2.CAR NK-92 cells) against the PD-L1-expressing MDA-MB-231 and ErbB2-expressing MDA-MB-453 breast cancer cell lines. Injected cancer cells were viable and populated peripheral regions of the larvae, including the caudal hematopoietic tissue (CHT), simulating homing of cancer cells to blood forming sites. CAR-NK cells injected 2.5 hours later migrated to the CHT and rapidly eliminated individual cancer cells throughout the organism. Unmodified NK-92 also demonstrated minor in vivo cytotoxicity. Confocal live-cell imaging demonstrated intravascular migration and real-time interaction of CAR-NK cells with MDA-MB-231 cells, explaining the rapid and effective in vivo cytotoxicity. Thus, our data suggest that zebrafish larvae can be used for rapid and cost-effective in vivo assessment of CAR-NK cell potency and to predict patient response to therapy.
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