《生命科学》 2022, 34(10): 1274-1283

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

张楫钦^1,* , 胡永仙² , 杨佳璇¹ , 李　伟³ , 李大力^1,* , 杜　冰^1,* , 刘明耀^1,* , 黄　河^2,*

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

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。

通讯作者：张楫钦 , Email:zjqjeremy@163.com 李大力 , Email:dlli@bio.ecnu.edu.cn 杜　冰 , Email:bdu@bio.ecnu.edu.cn 刘明耀 , Email:myliu@bio.ecnu.edu.cn 黄　河 , Email:huanghe@zju.edu.cn

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

ZHANG Ji-Qin^1,* , HU Yong-Xian² , YANG Jia-Xuan¹ , LI Wei³ , LI Da-Li^1,* , DU Bing^1,* , LIU Ming-Yao^1,* , HUANG He^2,*

¹East China Normal University, Shanghai 200241, China ²The First Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou 310003, China ³BRL Medicine Inc., Shanghai 201109, China

Abstract:

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.

Communication Author：ZHANG Ji-Qin , Email:zjqjeremy@163.com LI Da-Li , Email:dlli@bio.ecnu.edu.cn DU Bing , Email:bdu@bio.ecnu.edu.cn LIU Ming-Yao , Email:myliu@bio.ecnu.edu.cn HUANG He , Email:huanghe@zju.edu.cn