靶向胰腺癌的嵌合抗原受体T细胞U87在荷瘤小鼠体内的临床前毒性评价  

Preclinical Toxicity Evaluation of Chimeric Antigen Receptor T Cells U87 Targeting at Pancreatic Cancer in Tumor Bearing Mice

作　　者：文海若 霍桂桃[1] 秦超 赵晶 寇小旋 楼小燕 周晓冰[1] 黄瑛[1] 俞磊 WEN Hairuo;HUO Guitao;QIN Chao;ZHAO Jing;KOU Xiaoxuan;LOU Xiaoyan;ZHOU Xiaobing;HUANG Ying;YU Lei(Key Laboratory of Beijing for Nonclinical Safety Evaluation Research of Drugs,National Center for Safety Evaluation of Drugs,National Institutes for Food and Drug Control,Beijing 100176,China;Shanghai Unicar Therapy Biomedicine Technology Co.,Ltd.,Shanghai 201203,China)

机构地区：[1]中国食品药品检定研究院国家药物安全评价监测中心,药物非临床安全评价研究北京市重点实验室,北京100176 [2]上海优卡迪生物医药科技有限公司,上海201203

出　　处：《中国药学杂志》2024年第20期1889-1898,共10页Chinese Pharmaceutical Journal

基　　金：国家重点研发计划课题资助(2021YFA1101602);中国科学院战略先导科技专项资助(XDA1604050202);中国食品药品检定研究院关键技术研究基金资助(GJJS-2022-6-1)。

摘　　要：目的评价嵌合抗原受体T(chimeric antigen receptor T,CAR-T)细胞U87在荷瘤免疫缺陷小鼠体内的临床前安全性。方法使用NCG小鼠皮下接种人原位胰腺癌细胞BxPC-3构建移植瘤模型,并在此基础上单次给予细胞U87(每只动物2×10^(6)个和10×10^(6)个细胞),持续观察至给药后第84天。考察小鼠肿瘤清除情况、存活率、临床症状、体质量变化、血液学指标、血清生化指标检测、淋巴细胞亚群分类、人源细胞因子指标和组织病理学变化特点。结果单次尾静脉给予荷瘤NCG小鼠U87可明显延长小鼠生存期,甚至可完全清除肿瘤负荷。作为其药效学作用,U87可导致人源IFN-γ水平升高,以及多组织/脏器的混合细胞聚集,未见与U87相关的局部刺激性和明显系统毒性。U87在高剂量下可导致过度免疫反应,对动物GLU、TG、TP、ALB、PLT及RET数值产生一定影响。当体内肿瘤消除后,上述异常指标有所恢复。每只动物给予2×10^(6)个CAR-T(自给予细胞后第8周起)和10×10^(6)个CAR-T(自给予细胞后第4周起)剂量下均可导致轻度GvHD相关的组织病理学改变,且该变化与T细胞增殖水平相关联。未见与U87相关的肿瘤形成。结论U87在BxPC-3荷瘤NCG小鼠体内未见明显免疫毒性和致瘤性的相关风险,且呈现一定药效学作用。本研究结果为靶向实体瘤的毒理学评价提供借鉴。OBJECTIVE To evaluate the preclinical safety of chimeric antigen receptor T(CAR-T)cell U87 in tumor-bearing immunodeficient mice.METHODS A transplantation tumor model was constructed using NCG mice subcutaneously inoculated with BxPC-3,on which cells U87(2×10^(6)cells each mouse and 10×10^(6)cells each mouse)were administered in a single dose,and continued to be observed until day 84 after administration.Tumor clearance,survival rate,clinical symptoms,body weight changes,hematological indices,lymphocyte subpopulation classification,cytokine indices,and histopathological changes were examined to characterize the mice.RESULTS Single tail vein administration of U87 to hormonal NCG mice significantly prolonged mouse survival and even completely cleared the tumor load.As its pharmacodynamic action,U87 resulted in elevated levels of human-derived IFN-γ,as well as mixed cell aggregation in multiple tissues/organisms,with no local irritation and no apparent systemic toxicity associated with U87.U87 led excessive immune response at high doses,and has certain effects on the values of GLU,TG,TP,ALB,PLT and RET in animals.Mild GvHD-associated histopathological changes were only observed in 2×10^(6)CAR-T(from week 8 after cell administration)and 10×10^(6)CAR-T(from week 4 after cell administration),and the changes correlated with the level of T-cell proliferation.No tumor formation associated with U87 was seen.CONCLUSION U87 does not show any significant risk associated with immunotoxicity and tumorigenicity in BxPC-3 loaded NCG mice and shows some pharmacodynamic effects.The results of this study provide implications for the toxicological evaluation of targeted solid tumors.

关 键 词：嵌合抗原受体T细胞 胰腺癌 免疫缺陷荷瘤小鼠 实体瘤 毒性评价 

分 类 号：R979.1[医药卫生—药品]