降低亲和力提高HER2-CAR-T细胞治疗的安全性

章浩; 叶真龙; 钱其军

doi:10.3969/j.issn.1006-0111.2016.03.018

降低亲和力提高HER2-CAR-T细胞治疗的安全性

doi: 10.3969/j.issn.1006-0111.2016.03.018

第二军医大学东方肝胆外科医院病毒基因治疗实验室, 上海 200438

Decreasing affinity of CAR-T cells targeting HER2 to increase therapeutic outcome against tumors

Laboratory of Viral and Gene Therapy, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai 200438, China

摘要: 目的探讨降低CAR-T细胞亲和力是否能有效提高其杀伤特异性,减少"脱靶效应"。方法构建靶向HER2的中等亲和力和高亲和力的La-G3HER2-CAR和Ha-G3HER2-CAR并电穿孔转染T细胞,采用Western Blot、FCM技术和xCELLigence RTCA DP进行CAR载体表达和杀伤功能检测。结果 La-G3HER2-CAR-T细胞和Ha-G3HER2-CAR-T细胞分别表达43000和58000的外源CAR载体片段,转染效率分别为58.1%和69.0%。高亲和力的Ha-G3HER2-CAR-T细胞对高、中、低水平表达HER2的6种靶细胞均有效杀伤,而低亲和力的La-G3HER2-CAR-T细胞高效杀伤HER2高表达的SK-OV-3和BT474细胞,对HER2中表达的MDA-MB-231和HCC-202的杀伤作用较弱,而对低水平表达HER2的MCF-7和293细胞不杀伤。进一步的机制研究发现,HER2中水平表达的MDA-MB-231细胞共培养对La-G3HER2-CAR-T细胞和Ha-G3HER2-CAR-T细胞激活和细胞因子分泌诱导水平不同(CD107a:8.2% vs 71.6%; IFN-γ:66.3% vs 83.4%; TNF-α:73.4% vs 94.1%)。结论中等亲和力La-G3HER2-CAR-T细胞比高亲和力的Ha-G3HER2-CAR-T细胞的杀伤作用特异性更强,降低CAR-T细胞的亲和力可以提高治疗的安全性。
- 人表皮生长因子受体2 /
- 嵌合抗原受体 /
- T细胞 /
- 细胞免疫治疗
Abstract: Objective To investigate whether decreasing affinity of CAR-T cells can increase their therapeutic outcome or not. Methods Moderate affinity La-G3HER2-CAR and high affinity Ha-G3HER2-CAR were constructed, and electroporated to modify T cells. Western blot assay, FCM assay and the RTCA DP cytotoxic equipment were applied to test the CAR expression and cytotoxic function of CAR-T cells. Results 43000 and 58000 exogenous CD3ζ fragments were expressed by both La-G3HER2-CAR-T cells and Ha-G3HER2-CAR-T cells with 58.1% and 69.0% transfection rate respectively. High affinity Ha-G3HER2-CAR-T cells effectively killed all target tumor cells by which HER2 was expressed at variable expression levels, while moderate affinity La-G3HER2-CAR-T cells specifically killed HER2 high-level expressing SK-OV-3 and BT474 cells, and showed weaker cytotoxicity on HER2 moderate-level expressing MDA-MB-231 and HCC-202 cells, and showed no cytotoxicity on HER2 low-level expressing MCF-7 and 293 cells. The underlying mechanic investigation found that La-G3HER2-CAR-T cells and Ha-G3HER2-CAR-T cells were differentially activated by co-culture with MDA-MB-231(CD107a:8.2% vs 71.6%, IFN-γ:66.3% vs 83.4%, TNF-α:73.4% vs 94.1%). Conclusion Moderate affinity La-G3HER2-CAR-T cells have enhanced specific cytotoxicity toward target tumor cells compared to high affinity Ha-G3HER2-CAR-T cells, decreasing affinity of CAR-T cell is a promising strategy to increase the therapeutic outcome of CAR-T cell based immunotherapies.
- HER2 /
- chimeric antigen receptor /
- T cells /
- cellular immunotherapy

[1]	Maude SL, Frey N, Shaw PA, et al. Chimeric antigen receptor T cells for sustained remissions in leukemia[J]. N Engl J Med, 2014, 371(16):1507-1517.
[2]	Shen H, Laird PW. Interplay between the cancer genome and epigenome[J]. Cell, 2013, 153(1):38-55.
[3]	Roessler S, Budhu A, Wang XW. Deciphering cancer heterogeneity:the biological space[J]. Front Cell Dev Biol, 2014, 2:12.
[4]	Beavis PA, Slaney CY, Kershaw MH, et al. Reprogramming the tumor microenvironment to enhance adoptive cellular therapy[J]. Semin Immunol, 2015, 11:3.
[5]	Park HJ, Kusnadi A, Lee EJ, et al. Tumor-infiltrating regulatory T cells delineated by upregulation of PD-1 and inhibitory receptors[J]. Cell Immunol, 2012, 278(1):76-83.
[6]	Ciceri F, Bonini C, Stanghellini MT, et al. Infusion of suicide-gene-engineered donor lymphocytes after family haploidentical haemopoietic stem-cell transplantation for leukaemia (the TK007 trial):a non-randomised phase Ⅰ-Ⅱ study[J]. Lancet Oncol, 2009, 10(5):489-500.
[7]	Kloss CC, Condomines M, Cartellieri M, et al. Combinatorial antigen recognition with balanced signaling promotes selective tumor eradication by engineered T cells[J]. Nat Biotechnol, 2013, 31(1):71-75.
[8]	Caruso HG, Hurton LV, Najjar A, et al. Tuning sensitivity of CAR to EGFR density limits recognition of normal tissue while maintaining potent antitumor activity[J]. Cancer Res, 2015, 75(17):3505-3518.
[9]	Sun M, Shi H, Liu C, et al. Construction and evaluation of a novel humanized HER2-specific chimeric receptor[J]. Breast Cancer Res, 2014, 16(3):R61.
[10]	Lanitis E, Dangaj D, Hagemann IS, et al. Primary human ovarian epithelial cancer cells broadly express HER2 at immunologically-detectable levels[J]. PLoS One, 2012, 7(11):e49829.
[11]	Hapuarachchige S, Kato Y, Artemov D. Bioorthogonal two-component drug delivery in HER2(+) breast cancer mouse models[J]. Sci Rep, 2016, 6:24298.
[12]	Kulhari H, Pooja D, Shrivastava S, et al. Trastuzumab-grafted PAMAM dendrimers for the selective delivery of anticancer drugs to HER2-positive breast cancer[J]. Sci Rep, 2016, 6:23179.
[13]	左明辉, 金华君, 黎江, 等. 多靶向VEGF-EGFR的Fc融合蛋白EVP1的构建及其结合特性[J]. 中国肿瘤生物治疗杂志, 2012, 19(3):239-246.
[14]	Zhong XS, Matsushita M, Plotkin J, et al. Chimeric antigen receptors combining 4-1BB and CD28 signaling domains augment PI₃ kinase/AKT/Bcl-X_L activation and CD8⁺ T cell-mediated tumor eradication[J]. Mol Ther, 2010, 18(2):413-420.
[15]	Kenderian SS, Ruella M, Gill S, et al. Chimeric antigen receptor T-cell therapy to target hematologic malignancies[J]. Cancer Res, 2014, 74(22):6383-6389.
[16]	Harris DT, Kranz DM. Adoptive T cell therapies:A comparison of T cell receptors and chimeric antigen receptors[J]. Trends Pharmacol Sci, 2016, 37(3):220-230.

[1]	陈静, 何瑞华, 翁月, 徐熠, 刘静, 黄瑾. 基于网络药理学和分子对接技术探究定清片活性成分治疗白血病的作用机制 . 药学实践与服务, 2024, 42(): 1-8. doi: 10.12206/j.issn.2097-2024.202401073
[2]	丁华敏, 郭羽晨, 秦春霞, 宋志兵, 孙莉莉. 消风止痒颗粒通过降低白三烯水平对小鼠特应性皮炎急性瘙痒的治疗作用研究 . 药学实践与服务, 2024, 42(5): 211-216. doi: 10.12206/j.issn.2097-2024.202306031
[3]	陈春娟, 郑志新, 李骊. 平喘方联合孟鲁司特钠治疗儿童支气管哮喘患者的临床疗效观察 . 药学实践与服务, 2024, 42(): 1-5. doi: 10.12206/j.issn.2097-2024.202405035
[4]	景凯, 杨慈荣, 张圳, 臧艺蓓, 刘霞. 黄芪甲苷衍生物治疗慢性心力衰竭小鼠的药效评价及作用机制研究 . 药学实践与服务, 2024, 42(5): 190-197. doi: 10.12206/j.issn.2097-2024.202310004
[5]	孙丹倪, 黄勇, 张嘉宝, 王培. 代谢相关脂肪性肝病的无创诊断与药物治疗 . 药学实践与服务, 2024, 42(10): 411-418. doi: 10.12206/j.issn.2097-2024.202403049
[6]	张晶晶, 索丽娜, 郑兆红. 89例细菌性肝脓肿的临床特征及抗感染治疗分析 . 药学实践与服务, 2024, 42(6): 267-272. doi: 10.12206/j.issn.2097-2024.202302039
[7]	桂明珠, 李静, 李志玲. 儿童伏立康唑的血药浓度与CYP2C19、CYP2C9和CYP3A5基因多态性的相关性研究 . 药学实践与服务, 2024, 42(): 1-5. doi: 10.12206/j.issn.2097-2024.202402020
[8]	陈莹, 许子华, 胡北, 崔亚玲, 高欢, 吴琼. 通便灵胶囊治疗便秘的药效与机制研究 . 药学实践与服务, 2024, 42(): 1-7. doi: 10.12206/j.issn.2097-2024.202404008
[9]	宋泽成, 陈林林, 鲁仁义, 刘梦肖, 王彦. 脓毒症治疗的研究进展 . 药学实践与服务, 2024, 42(11): 1-5. doi: 10.12206/j.issn.2097-2024.202405059
[10]	顾佳钰, 胡馨儿, 王晓飞, 张颖, 张海, 曹岩. 侧流免疫层析定量检测方法的研究进展 . 药学实践与服务, 2024, 42(7): 273-277, 284. doi: 10.12206/j.issn.2097-2024.202307037
[11]	毛智毅, 王筱燕, 陈晓颖, 汤逸斐. 度拉糖肽联合二甲双胍对肥胖型2型糖尿病患者机体代谢、体脂成分及血清脂肪因子的影响 . 药学实践与服务, 2024, 42(7): 305-309. doi: 10.12206/j.issn.2097-2024.202305032
[12]	唐淑慧, 凤美娟, 薛智霞, 鲁桂华. 帕博利珠单抗治疗所致免疫相关不良反应与中医体质的相关性研究 . 药学实践与服务, 2024, 42(5): 217-222. doi: 10.12206/j.issn.2097-2024.202311029
[13]	冯志惠, 邓仪卿, 叶冰, 安培, 张宏, 张海军. 雀梅藤石油醚提取物诱导三阴性乳腺癌细胞凋亡的实验研究 . 药学实践与服务, 2024, 42(6): 253-259. doi: 10.12206/j.issn.2097-2024.202311055
[14]	修建平, 杨朝爱, 刘禧澳, 潘乾禹, 韦广旭, 王卫星. 全反式维甲酸对肝星状细胞活化及氧化应激的作用和机制探索 . 药学实践与服务, 2024, 42(7): 291-296. doi: 10.12206/j.issn.2097-2024.202312054
[15]	姜涛, 徐卫凡, 蒋益萍, 夏天爽, 辛海量. 巴戟天丸组方对Aβ损伤成骨细胞的作用及基于网络药理学的机制研究 . 药学实践与服务, 2024, 42(7): 285-290, 296. doi: 10.12206/j.issn.2097-2024.202305011
[16]	刘丽艳, 余小翠, 孙传铎. 纳武利尤单抗治疗非小细胞肺癌有效性及安全性的Meta分析 . 药学实践与服务, 2024, 42(10): 451-456. doi: 10.12206/j.issn.2097-2024.202310044
[17]	迟文雅, 袁艳, 李伟林, 吴茼妤, 俞媛. 负载骨髓间充质干细胞/白藜芦醇脂质体的水凝胶支架用于创伤性脑损伤治疗 . 药学实践与服务, 2024, 42(): 1-8. doi: 10.12206/j.issn.2097-2024.202406034
[18]	宋雨桐, 夏德润, 顾珩, 唐少文, 易洪刚, 沃红梅. 帕博利珠单抗与铂类化疗方案在晚期非小细胞肺癌一线治疗中的药物经济学评价 . 药学实践与服务, 2024, 42(8): 334-340. doi: 10.12206/j.issn.2097-2024.202303023
[19]	杨媛媛, 安晓强, 许佳捷, 江键, 梁媛媛. 正极性驻极体联合5-氟尿嘧啶对瘢痕成纤维细胞生长抑制的协同作用 . 药学实践与服务, 2024, 42(6): 244-247. doi: 10.12206/j.issn.2097-2024.202310027
[20]	马兹芬, 许维恒, 金煜翔, 薛磊. 食管癌的靶向治疗与免疫治疗研究进展 . 药学实践与服务, 2024, 42(6): 231-237. doi: 10.12206/j.issn.2097-2024.202306008

点击查看大图

计量

文章访问数: 3647
HTML全文浏览量: 471
PDF下载量: 24
被引次数: 0

姓名
邮箱
手机号码
标题
留言内容
验证码

留言板

降低亲和力提高HER2-CAR-T细胞治疗的安全性

doi: 10.3969/j.issn.1006-0111.2016.03.018

Decreasing affinity of CAR-T cells targeting HER2 to increase therapeutic outcome against tumors

计量

降低亲和力提高HER2-CAR-T细胞治疗的安全性

doi: 10.3969/j.issn.1006-0111.2016.03.018

第二军医大学东方肝胆外科医院病毒基因治疗实验室, 上海 200438

English Abstract

Decreasing affinity of CAR-T cells targeting HER2 to increase therapeutic outcome against tumors

Laboratory of Viral and Gene Therapy, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai 200438, China

全文HTML

目录

留言板

降低亲和力提高HER2-CAR-T细胞治疗的安全性

doi: 10.3969/j.issn.1006-0111.2016.03.018

Decreasing affinity of CAR-T cells targeting HER2 to increase therapeutic outcome against tumors

计量

出版历程

降低亲和力提高HER2-CAR-T细胞治疗的安全性

doi: 10.3969/j.issn.1006-0111.2016.03.018

第二军医大学东方肝胆外科医院病毒基因治疗实验室, 上海 200438

English Abstract

Decreasing affinity of CAR-T cells targeting HER2 to increase therapeutic outcome against tumors

Laboratory of Viral and Gene Therapy, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai 200438, China

全文HTML

目录