Engineering a controllable and reversible switch for CAR-based cellular immunotherapies via a genetic code expansion...

Area of Science:

• Biotechnology
• Immunotherapy
• Synthetic Biology

Background:

• Chimeric antigen receptor T (CAR-T) cell therapy shows promise for hematological tumors but faces challenges like toxicity and recurrence.
• Current CAR-T therapy lacks precise control over protein expression and function, limiting its safety and efficacy.
• There is a critical need for strategies to enhance the controllability and safety of CAR-T cell therapies.

Purpose of the Study:

• To engineer a genetic code expansion system for controllable CAR protein expression and regulation.
• To investigate the use of an unnatural amino acid, BOCK, as a switch to control CAR-T cell function.
• To evaluate the safety and efficacy of this BOCK-induced system in vitro and in vivo.

Main Methods:

• Engineered a genetic code expansion system using the unnatural amino acid BOCK to regulate CAR protein translation.
• Developed a system where CAR protein expression is "closed" without BOCK and "open" with BOCK.
• Conducted in vitro and in vivo experiments to assess CAR-T cell function and anti-tumor activity.

Main Results:

• Verified the BOCK-induced system as a controllable switch for protein expression.
• Demonstrated precise control over CAR protein expression and CAR-T cell signaling activation by BOCK.
• Showcased dose-dependent regulation of CAR-T cell cytotoxicity against tumor cells and significant anti-tumor effects in a mouse model.
• Confirmed the universality of the BOCK-induced device for controlling NK cell anti-tumor activity.

Conclusions:

• The BOCK-induced genetic code expansion system precisely regulates CAR protein expression and controls CAR-T cell anti-tumor effects.
• This controllable and reversible switch offers potential for safer and more effective CAR-based immunotherapies.
• The system holds promise for enhanced clinical applications of CAR-based cellular immunotherapies.