Synthetic biology approaches to improve cell-based cancer immunotherapy

Adoptive T cell immunotherapy is a promising anti-cancer therapy that has the potential to become the ultimate therapeutic agents to treat a variety of diseases. Recently, chimeric antigen receptor (CAR) expressing T cells has demonstrated to be a very effective approach to treat B cell cancer patients. Despite optimistic results, there are several improvements that need to be made to enhance the safety and efficacy of current CAR T cell therapy. Fortunately, different synthetic biology tools can be implemented to overcome many of the current deficiencies of CAR T cell therapy. Here, we develop anti-Axl CAR and synNotch receptors to target Axl which is a tyrosine kinase receptor that is commonly overexpressed in many cancers and considered as one of important cancer therapeutic targets. Next, we develop a split, universal, and programmable (SUPRA) CAR system that can be used to switch targets without re-engineering T cells, fine-tune T cell activation level, and sense and logically respond to multiple antigens. These multiple features are useful in mitigating tumor relapse, limiting CAR-T induced toxicities, and enhancing tumor specificity. Orthogonal SUPRA CARs are also used to control different cell types and signaling domains, enabling diverse immune responses from SUPRA CAR T cells. Lastly, we demonstrate that SUPRA CAR can redirect the activity of both innate and adaptive immune cell types. We also expand the logic capabilities of SUPRA CAR T cells by integrating three inputs in a single immune cell. We also show intercellular logic gate by engineering immune cell-cell interaction. We further demonstrate controlling endogenous immune cell polarization using SUPRA CAR T cells. These wide-ranges of SUPRA CAR applications imply its versatility as a platform for engineering cell-cell interactions with advanced logic functions to enhance efficacy and safety of cell-based cancer immunotherapy