Synthetic Biology and Biomedical Engineering Laboratory, Biomedical Synthetic Biology Research Center, Shanghai Key Laboratory of Regulatory Biology, Institute of Biomedical Sciences and School of Life Sciences, East China Normal University, Shanghai 200241, China.
Wyss Institute for Biologically Inspired Engineering, Harvard University, Boston, MA 02215.
Proc Natl Acad Sci U S A. 2021 Aug 24;118(34). doi: 10.1073/pnas.2106612118.
Chimeric antigen receptor (CAR)-engineered T cell therapies have been recognized as powerful strategies in cancer immunotherapy; however, the clinical application of CAR-T is currently constrained by severe adverse effects in patients, caused by excessive cytotoxic activity and poor T cell control. Herein, we harnessed a dietary molecule resveratrol (RES)-responsive transactivator and a transrepressor to develop a repressible transgene expression (RES) device and an inducible transgene expression (RES) device, respectively. After optimization, these tools enabled the control of CAR expression and CAR-mediated antitumor function in engineered human cells. We demonstrated that a resveratrol-repressible CAR expression (RES-CAR) device can effectively inhibit T cell activation upon resveratrol administration in primary T cells and a xenograft tumor mouse model. Additionally, we exhibit how a resveratrol-inducible CAR expression (RES-CAR) device can achieve fine-tuned and reversible control over T cell activation via a resveratrol-titratable mechanism. Furthermore, our results revealed that the presence of RES can activate RES-CAR T cells with strong anticancer cytotoxicity against cells in vitro and in vivo. Our study demonstrates the utility of RES and RES devices as effective tools for transgene expression and illustrates the potential of RES-CAR and RES-CAR devices to enhance patient safety in precision cancer immunotherapies.
嵌合抗原受体 (CAR)-修饰的 T 细胞疗法已被认为是癌症免疫疗法的有力策略；然而，CAR-T 的临床应用目前受到患者中严重不良反应的限制，这是由过度的细胞毒性活性和 T 细胞控制不良引起的。在此，我们利用膳食分子白藜芦醇（RES）响应性转录激活子和转录阻遏子分别开发了一种可抑制的转基因表达（RES）装置和一种诱导型转基因表达（RES）装置。经过优化，这些工具能够控制工程化人细胞中的 CAR 表达和 CAR 介导的抗肿瘤功能。我们证明，白藜芦醇可抑制的 CAR 表达（RES-CAR）装置可在原发性 T 细胞和异种移植肿瘤小鼠模型中在给予白藜芦醇后有效抑制 T 细胞活化。此外，我们展示了如何通过白藜芦醇滴定机制实现通过 RES 诱导的 CAR 表达（RES-CAR）装置实现对 T 细胞活化的精细和可逆控制。此外，我们的结果表明，RES 的存在可以激活 RES-CAR T 细胞，对体外和体内的癌细胞具有强大的抗癌细胞毒性。我们的研究证明了 RES 和 RES 装置作为转基因表达的有效工具的实用性，并说明了 RES-CAR 和 RES-CAR 装置在精确癌症免疫疗法中提高患者安全性的潜力。
