Targeting CD20+ Aggressive B-cell Non-Hodgkin Lymphoma by Anti-CD20 CAR mRNA-Modified Expanded Natural Killer Cells In Vitro and in NSG Mice
Targeting CD20+ Aggressive B-cell Non-Hodgkin Lymphoma by Anti-CD20 CAR mRNA-Modified Expanded Natural Killer Cells In Vitro and in NSG Mice
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DOI：
10.1158/2326-6066.cir-14-0114
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发表时间：
2015-04-01
影响因子：
10.1
通讯作者：
Cairo, Mitchell S.
中科院分区：
文献类型：
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作者：
Chu, Yaya;Hochberg, Jessica;Cairo, Mitchell S.
复发的CD20(+) b细胞非霍奇金淋巴瘤（B-NHL）患者的预后非常糟糕。促进替代性新治疗策略的发展是改善复发/难治性CD20(+) B-NHL患者预后的必要条件。在这项研究中，我们在体外和体内研究了抗CD20 car修饰的扩增外周血NK细胞（exPBNK）在mRNA核感染后对CD20(+) B-NHL的功能活性。与CAR(-) exPBNK相比，CAR(+) exPBNK对CD20(+) Ramos （P < 0.05）、Daudi、Raji和两种利妥昔单抗耐药细胞系Raji- 2r和Raji- 4rh的体外细胞毒性显著增强（P < 0.001）。不出所料，对CD20(-) RS4无显著性差异；11和Jurkat细胞。CAR(+) exPBNK在CD20(+) b - nhl特异性刺激下也增强了CD107a脱颗粒和细胞内IFN γ的产生。在Raji-Luc和Raji-2R-Luc异种移植NOD/SCID/ γ -链（-/-）（NSG）小鼠中，与未治疗小鼠和CAR(-) exPBNK处理小鼠相比，CAR(+) exPBNK处理组测得的荧光素酶信号显著降低。此外，与未治疗和CAR(-) expbnk处理的小鼠相比，CAR(-) expbnk处理的小鼠生存时间显著延长（P < 0.001），肿瘤大小显著减小（P < 0.05）。这些临床前数据表明，抗CD20 CAR修饰的体外expbnk可能具有治疗低风险CD20（+）血液恶性肿瘤患者的治疗潜力。
The prognosis is very dismal for patients with relapsed CD20(+) B-cell non-Hodgkin lymphoma (B-NHL). Facilitating the development of alternative novel therapeutic strategies is required to improve outcomes in patients with recurrent/refractory CD20(+) B-NHL. In this study, we investigated functional activities of anti-CD20 CAR-modified, expanded peripheral blood NK cells (exPBNK) following mRNA nucleofection against CD20(+) B-NHL in vitro and in vivo. CAR(+) exPBNK had significantly enhanced in vitro cytotoxicity, compared with CAR(-) exPBNK against CD20(+) Ramos (P < 0.05), Daudi, Raji, and two rituximab-resistant cell lines, Raji-2R and Raji-4RH (P < 0.001). As expected, there was no significant difference against CD20(-) RS4; 11 and Jurkat cells. CD107a degranulation and intracellular IFN gamma production were also enhanced in CAR(+) exPBNK in response to CD20(+) B-NHL-specific stimulation. In Raji-Luc and Raji-2R-Luc xenografted NOD/SCID/gamma-chain(-/-) (NSG) mice, the luciferase signals measured in the CAR(+) exPBNK-treated group were significantly reduced, compared with the signals measured in the untreated mice and in mice treated with the CAR(-) exPBNK. Furthermore, the CAR exPBNK-treated mice had significantly extended survival time (P < 0.001) and reduced tumor size, compared with those of the untreated and the CAR(-) exPBNK-treated mice (P < 0.05). These preclinical data suggest that ex vivo-exPBNK modified with anti-CD20 CAR may have therapeutic potential for treating patients with poor-risk CD20(+) hematologic malignancies.