CRISPR - Activation and knockout screens to augment anti-tumor T cell responses

Immunotherapy is proving to be an effective therapeutic approach to treat cancer. However, the remarkable responses to immunotherapies are currently limited to a minority of patients and indications, highlighting the need to identify additional targets and to develop novel and more effective approaches. In this project, we aim to perform pooled CRISPR-activation and CRISPR-knockout screens in primary human T cells to identify the genes that govern effective T cell responses to tumors. We identified a list of ~3,000 metabolic genes, which will be either deleted or activated in T cells to reveal casual relationships with anti-tumor functionality. In Aim 1, we will perform pooled CRISPR screens to identify genes that affect the effector functions of T cells under metabolically challenging conditions found in tumors. In Aim 2, we will use droplet-based microfluidics systems to develop a high-throughput T cell killing assay that allows us to perform pooled CRISPR screens for the identification of genes that influence the cytotoxic capacity of T cells. In Aim 3, we will validate a subset of “candidate genes” identified Aim 1 and 2 in both human T cells as well as in murine tumor models. This proposal combines state-of-the-art genetic screening with innovative methods to analyze biologically meaningful T cell phenotypes and has the potential to reveal new targets for immunotherapy.