Novel Approach to High-Throughput Identification and Characterization of Neoantigen-Specific T-Cell Receptors to Guide Immunotherapy Development.

PROJECT SUMMARY Newly developed immunotherapies have shown great promise for the management and treatment of various types of cancer. Among these new strategies, the development of T-cell Receptor engineered T-cells (TCR T- cells) targeting tumor specific neoantigens has arisen as a promising approach to improve efficacy, decrease toxicity and overcome acquired resistance. However, the multiplicity of mutations in cancer and the lack of high- throughput methods to identify neoantigen-specific TCRs is preventing the wide implementation of TCR T-cells therapies. Flexomics is developing an integrated platform harnessing recent advances in single cell genomics, immunoassays and microfluidics for the high-throughput characterization of antigen-specific TCRs. Our proprietary approach is designed to identify T-cell activation in response to specific antigen exposure at single cell level for 100,000s T-cells while simultaneously capturing phenotypic and genotypic information in the form of expression of key marker genes and full-length TCR sequence identification. To demonstrate the feasibility of our method we propose to: (1) track functional responses from single T-cells to cognate antigen-exposure by measuring their cytokine secretion in our novel high-density picowell plate that allows us to co-capture 100,000s pairs of Antigen Presenting Cell (APC) / T-cell, (2) retrieve and link TCR sequence and expression for 20 phenotypic markers to each individual T-cell thanks to our unique barcoding system, (3) demonstrate end-to-end workflow and recapitulate previously discovered antigen-specific TCR sequence for a well characterized neoantigen. At the end of our Phase I grant, we will have demonstrated a concrete example of how our platform allows the screening and characterization of antigen-TCR interactions at unprecedented scale. We will also have all processes in place to start screening other neoantigens and further refine our workflow. Screening and discovery of neoantigen-specific T-cell response and TCR is critical to the future progress of cancer immunotherapies. Our technology has been developed specifically to support and accelerate the development of personalized treatments by providing a rapid and cost-effective method to identify immunogenic neoantigens and characterize their associated antigen-specific TCRs.PROJECT NARRATIVE Identifying and characterizing neoantigen-specific TCR in a high-throughput manner is challenging. The proposed technology harnesses gold-standard immunoassays and cutting-edge single cell genomic technologies to meet this critical need and accelerate the development of personalized immunotherapies.