PT  - JOURNAL ARTICLE
AU  - Hui-Shan Li
AU  - Nicole M. Wong
AU  - Elliot Tague
AU  - John T. Ngo
AU  - Ahmad S. Khalil
AU  - Wilson W. Wong
TI  - Engineering clinically-approved drug gated CAR circuits
AID  - 10.1101/2020.12.14.419812
DP  - 2020 Jan 01
TA  - bioRxiv
PG  - 2020.12.14.419812
4099  - http://biorxiv.org/content/early/2020/12/14/2020.12.14.419812.short
4100  - http://biorxiv.org/content/early/2020/12/14/2020.12.14.419812.full
AB  - Chimeric antigen receptor (CAR) T cell immunotherapy has the potential to revolutionize cancer medicine. However, excessive CAR activation, lack of tumor-specific surface markers, and antigen escape have limited the safety and efficacy of CAR T cell therapy. A multi-antigen targeting CAR system that is regulated by safe, clinically-approved pharmaceutical agents is urgently needed, yet only a few simple systems have been developed, and even fewer have been evaluated for efficacy in vivo. Here, we present NASCAR (NS3 ASsociated CAR), a collection of induc-ible ON and OFF switch CAR circuits engineered with a NS3 protease domain deriving from the Hepatitis C Virus (HCV). We establish their ability to regulate CAR activity using multiple FDA-approved antiviral protease inhibitors, including grazoprevir (GZV), both in vitro and in a xenograft tumor model. In addition, we have engineered several dual-gated NASCAR circuits, consisting of an AND logic gate CAR, universal ON-OFF CAR, and a switchboard CAR. These engineered receptors enhance control over T cell activity and tumor-targeting specificity. Together, our com-prehensive set of multiplex drug-gated CAR circuits represent a dynamic, tunable, and clinically-ready set of modules for enhancing the safety of CAR T cell therapy.Competing Interest StatementThe authors have declared no competing interest.