Synthetic Mechano-Transduction For Improved Cell Therapies In Immuno-Oncology

ABSTRACT The utilization of genetically engineered T-cells containing Chimeric Antigen Receptors (CARs) has demonstrated remarkable efficacy in treating hematologic malignancies. Despite this success, CAR-T cell therapies have encountered challenges in specificity and off-target toxicity when confronting solid tumors. To address these challenges, we propose an innovative approach that harnesses mechanical forces, which is overlooked in current therapeutic strategies. Mechanical forces are pivotal in immune signaling because they impact the activation of key receptors, such as the T cell receptor, chimeric antigen receptor (CAR), and co- receptor PD1. We recently developed force sensitive coiled-coils that can be genetically encoded and inserted into any protein of interest. These coiled-coils change conformation under force and expose reactive residues that can be harnessed for force detection or enzyme recruitment. This project aims to build on our experience to engineer a mechano-transduction platform that will be implemented to enhance CAR T-cell therapies. In aim 1, we will use our force sensors to measure the forces along key immune receptors. In aim 2, we will engineer force transduction modules by combining our force sensors to releasable transcription activators able to induce the expression of any protein of choice, and insert these tools into CARs to enhance CAR T-cell specificity and safety. The successful realization of this project will demonstrate the feasibility and utility of new capabilities to improve the efficacy of CAR-T therapies, extend their application and speed up their development, with high potential impact in cancer immuno-therapies. It will also lay the foundations for new immuno-therapies using different receptors and modes of action.