Engineering genetic devices for in vivo control of therapeutic T cell activity triggered by the dietary molecule resveratrol

Abstract

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 (RESrep) device and an inducible transgene expression (RESind) 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 (RESrep-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 (RESind-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 RESind-CAR T cells with strong anticancer cytotoxicity against cells in vitro and in vivo. Our study demonstrates the utility of RESrep and RESind devices as effective tools for transgene expression and illustrates the potential of RESrep-CAR and RESind-CAR devices to enhance patient safety in precision cancer immunotherapies.

Prospective approaches to enhancing CAR T cell therapy for glioblastoma

“…Duong’s research group developed dual-switch CAR T cells in which one is a potent activation switch based on rimiducid-inducible MyD88 and CD40 (iMC)-signaling elements to improve CAR T cell efficacy, and the other is an orthogonally regulated, rapamycin-induced, caspase-9-based safety switch (iRC9) to neutralize potential toxicity by this reinforced CAR ( 55 ). Similarly, Yang’s research group constructed two types of CAR T cells with the stilbenoid natural product resveratrol (3,4’,5-trihy-droxystilbene) as a switch molecule, containing gene circuits that can control the activation (on) and inactivation (off) of CAR T cells ( 56 ). These tunable dual-switch systems provide a safety switch to reduce toxicity, thus supporting greater CAR T cell expansion and long-term persistence in a drug-dependent manner.…”

Application and Design of Switches Used in CAR

“…If resveratrol is absent, ResR1 blocks PhCMV by binding to OTRC1 and OTRC2. When resveratrol is administered, ResR1 is released and PhCMV is activated, which is associated with the activation of transcription of the desired gene [ 63 ].…”

CAR-T Immunotherapy to Beat Solid Tumors: From Challenges to Improvements

“…However, limiting CAR-T cell function by a switch is full of promise in circumventing side effects. Recent studies have demonstrated a transgenic expression system regulated by resveratrol, which regulates the expression (ON) and suppression (OFF) of CAR in T cells by resveratrol, leading to controlled CAR-T cell immunotherapy and improved safety of tumor immunotherapy [34]. Another study controlled the viability of CAR-T somatic cells in solid tumors by using focused ultrasound, which could lessen the off-target effect and reduce the harm of CAR-T to all normal structures [35] (Fig.…”