非病毒定点整合CAR-T技术的开发及其在复发难治性非霍奇金B细胞淋巴瘤临床治疗中的应用  

Development of non-viral genome specific targeted CAR-T technology and its application in clinical treatment of relapsed/refractory B-cell non-Hodgkin lymphoma

作　　者：张楫钦 胡永仙[2] 杨佳璇 李伟 李大力[1] 杜冰[1] 刘明耀[1] 黄河[2] ZHANG Ji-Qin;HU Yong-Xian;YANG Jia-Xuan;LI Wei;LI Da-Li;DU Bing;LIU Ming-Yao;HUANG He(East China Normal University,Shanghai 200241,China;The First Affiliated Hospital,Zhejiang University School of Medicine,Hangzhou 310003,China;BRL Medicine Inc.,Shanghai 201109,China)

机构地区：[1]华东师范大学,上海200241 [2]浙江大学医学院附属第一医院,杭州310003 [3]上海邦耀生物科技有限公司,上海201109

出　　处：《生命科学》2022年第10期1274-1283,共10页Chinese Bulletin of Life Sciences

基　　金：国家自然科学基金重点项目(2019YFA0802802,2018YFA0507001);国家自然科学基金面上项目(81772622,81730008,81770201,91857116,31871453)。

摘　　要：近年来,嵌合抗原受体(chimeric antigen receptor,CAR)T细胞疗法在治疗恶性血液肿瘤中取得了喜人的进展,但目前CAR-T疗法依然存在一定的问题。本课题组利用CRISPR/Cas9基因编辑系统成功开发了非病毒定点整合CAR-T技术。通过各种条件的摸索和优化,成功制备了靶向CD19的非病毒PD1定点整合型CAR-T细胞。临床前和临床研究结果显示,该CAR-T细胞在复发难治性非霍奇金B细胞淋巴瘤的治疗中具有出色的安全性和有效性。机制研究表明,该PD1定点整合CAR-T细胞具有更高比例的记忆性T细胞和更强的抗肿瘤免疫功能。本研究开发的非病毒定点整合CAR-T技术为解决现有CAR-T治疗领域存在的问题提供了新的方法和思路。Recently,chimeric antigen receptor(CAR)T cell therapy has shown great promise in treating hematological malignancies.However,CAR-T cell therapy currently has several limitations.Here we successfully developed an approach to generate non-viral genome specific targeted CAR-T cells through CRISPR/Cas9.Based on the optimized protocol,anti-CD19 CAR-T cells with PD1-integration were developed without using virus.The preclinical and clinical studies showed that this kind of CAR-T cells had high safety and efficacy in treating B-cell non-Hodgkin lymphoma.The mechanistic study indicated that these PD1-integrated CAR-T cells have a high percentage of memory T cells and enhanced anti-tumor immune functions.Collectively,this study developed a nonviral genome specific targeted CAR-T technology,thus providing a novel method and strategy to solve the current problems in CAR-T cell therapy.

关 键 词：CRISPR/Cas9基因编辑 非霍奇金B细胞淋巴瘤 非病毒定点整合CAR-T 安全性和有效性 复发难治 

分 类 号：Q343[生物学—遗传学] R730.51[医药卫生—肿瘤]