Greenberg, Philip D.Lee, Jihoon William2024-04-262024-04-262024Lee_washington_0250E_26559.pdfhttp://hdl.handle.net/1773/51382Thesis (Ph.D.)--University of Washington, 2024Adoptive T cell therapy (ACT) has emerged as a promising type of immunotherapy with clinical success against a variety of cancers. Generating T cells for ACT consists of isolation/generation, engineering with receptors such as T cell receptors (TCRs) and chimeric antigen receptors (CARs) to target a cancer antigen, and ex vivo expansion of T cells before they are reinfused into individuals with cancer. However, ACT faces challenges such as metabolic deficiencies, epigenetic programming, and inadequate recognition of tumor antigen that contribute to incomplete responses and possible relapse of tumors. In this work, I address these issues with approaches that should prove useful as complementary solutions to improve the overall long-term, tumor-targeting activity of ACT. First, I characterized the heterogenous metabolic changes that occur in immune cells in response to COVID-19. This opened insight into metabolic mechanisms that are potentially targetable to improve immune function in response to infection and tumor burden. Second, by examining the epigenetic and transcriptional programs potentially driving T cell dysfunction, I identified and targeted key epigenetic mechanisms, specifically the enforcement of early TGFβ signaling and inhibition of histone demethylase LSD1, during ex vivo therapeutic product generation to enhance the persistence and long-term antitumor activity of the generated T cells. These interventions conferred functional benefit when applied only during the ex vivo product generation, which increases potential clinical applicability. Third, I investigated the presence of posttranslational modifications on HLA-A2-presented KRASG12V mutant epitope, which is a highly common cancer-specific antigen and thus an attractive target for ACT but has been difficult to develop a TCR against. I found that methylation of the predicted epitope by tumor cells affects recognition by TCRs, which may be a mechanism by which tumor cells may evade immunologic pressure, and a TCR that recognizes the methylated peptide proved more effective than one that did not. Addressing the impact of such modifications of candidate epitope targets in TCR development should provide insight into designing more effective TCRs for TCR-T cell therapies.application/pdfen-USnoneAdoptive cell therapyCOVID-19EpigeneticsOnco-immunologyPosttranslational modificationT cell receptorMedicineImmunologyBiologyMolecular and cellular biologyThesis