设计、构建白藜芦醇调控的基因开关系统及其在肿瘤治疗中的应用研究

项目名称： 设计、构建白藜芦醇调控的基因开关系统及其在肿瘤治疗中的应用研究

项目编号： No.31670869

项目类型： 面上项目

立项/批准年度： 2017

项目学科： 心理学

项目作者： 叶海峰

作者单位： 华东师范大学

项目金额： 25万元

中文摘要： 白藜芦醇（RES）是具有抗肿瘤、抗炎症、抗衰老等多种生物活性的天然药物。因此，设计合成用于肿瘤治疗的RES调控系统具有极大意义。目前CAR-T细胞治疗白血病在临床上取得巨大成功。但CAR-T的不可控性导致严重的毒副作用，阻碍了CAR-T治疗的广泛应用。因此，本研究旨在探索利用合成生物学方法设计的RES调控系统调控CAR-T细胞的可行性，评估其用于规避传统方式毒副作用及治疗白血病的潜力。首先利用合成生物学的理念，设计合成RESON和RESOFF 两种基因开关；其次，对RES调控的两种基因开关分别在细胞水平和动物水平研究其调控基因表达的动力学特征；最后，利用该基因开关设计两种可控的CAR-T细胞并初步在细胞水平研究RES调控CAR-T杀伤白血病细胞的能力。该研究可为下一代精准可控、更安全有效的CAR-T细胞治疗提供新方法和新策略。

中文关键词： 合成生物学；调控元器件；设计；调控；生物器件

英文摘要： Resveratrol (RES) is a famous natural product which has been considered to play physiological roles such as anticancer, anti-inflammatory and anti-aging effects. So, it is of great significance to design RES-inducible transgene expression device for tumor therapy. The adoptive transfer of synthetic Chimeric Antigen Receptor (CAR) T cells has shown tremendous promise for eradicating leukemia in clinical trials. However, it is hard to be widely applied as such engineered T cells can cause severe toxicities because of their excessive activities and poor control. In this project, we use a synthetic biology strategy for controlling CAR-T cells by designing and creating RES-controlled devices. In addition, we want to confirm the potential benefits of this system for mitigating toxicity and curing leukemia. We first design and validate the RESON and RESOFF orthogonal gene switches and then we explore transgene expression kinetics of these two gene switches in mammalian cells and mice. Finally, we design two types of RES-controlled CAR-T cells based on these two gene switches and we preliminary test the therapeutic activity to kill leukemic cells of the controllable CAR-T cells induced by varying concentration of resveratrol in vitro. This approach will provide a new therapeutic strategy for CAR-T cellular immunotherapy which is more precise, controllable and safer.

英文关键词： Synthetic biology;Controllable components ;Rational design ;Gene regulation ;Biological device