000005220 001__ 5220 000005220 005__ 20251007025228.0 000005220 0247_ $$2doi$$a10.6082/uchicago.5220 000005220 037__ $$aTHESIS$$bDissertation 000005220 041__ $$aeng 000005220 245__ $$aIdentifying, Investigating, and Modulating Anti-CD19 CAR T Cells for Large B-Cell Lymphoma 000005220 260__ $$bUniversity of Chicago 000005220 269__ $$a2022-12 000005220 300__ $$a164 000005220 336__ $$aDissertation 000005220 502__ $$bPh.D. 000005220 520__ $$aAnti-CD19 chimeric antigen receptor (CAR) T-cell therapy has improved treatment options for patients with relapsed/refractory diffuse large B-cell lymphoma (r/r DLBCL). During anti-CD19 CAR T-cell therapy, the patient’s T cells are transduced with an anti-CD19 CAR – a synthetic immunoreceptor that directs the T cell to recognize, activate, proliferate, and kill in response to CD19+ lymphoma cells. Despite encouraging ~40% complete response rates, non-responders generally succumb to their disease. Hence, improved CAR T-cell therapy formulations with higher response rates are urgently needed in the clinic. My research questions, experiments, findings, and conclusions in this dissertation are organized around this central clinical problem. In this body of work, after providing a brief introduction to CD19-directed CAR T-cell therapy (Section I), I first reviewed the existing methods used by CAR T-cell scientists to detect and investigate the CAR across the genomic, transcriptomic, proteomic, and organismal levels (Section II). With this background in mind, I designed, constructed, and validated antigen multimers as specific, sensitive, precise, and multifunctional high-avidity reagents to detect the CAR at the proteomic level (Section III). Using antigen multimers to sort CAR T cells from patient biospecimens for longitudinal, single-cell, multi-omics analysis, I next reported that CD28-costimulated CAR T cells undergo two distinct clonal expansion stages in vivo, each of which is dominated by unique phenotypes (Section IV). Finally, using findings from comparing the single-cell data from complete responders and non-responders, I leveraged the type I interferon/interferon response factor 7 pathway for increasing CAR T-cell efficacy in vitro (Section V). Collectively, these findings are expected to introduce a better CAR-detection reagent for CAR T-cell scientists, improve how we understand CAR T-cell expansion and persistence, and synergize CAR T-cell therapy with an existing pharmaceutical in vitro. Ultimately, I hope these efforts will translate into improved CAR T-cell therapy formulations and longer progression-free survival for patients with r/r DLBCL. 000005220 540__ $$a© 2022 Yifei Hu 000005220 542__ $$fCC BY-NC-ND 000005220 650__ $$aImmunology 000005220 650__ $$aBiology 000005220 650__ $$aBioengineering 000005220 653__ $$aantigen multimer 000005220 653__ $$achimeric antigen receptor 000005220 653__ $$ainterferon 000005220 653__ $$alymphoma 000005220 653__ $$asequencing 000005220 653__ $$atwo-stage differentiation 000005220 690__ $$aBiological Sciences Division 000005220 690__ $$aPritzker School of Medicine 000005220 691__ $$aInterdisciplinary Scientist Training Program 000005220 7001_ $$aHu, Yifei$$uUniversity of Chicago 000005220 72012 $$aJun Huang 000005220 72014 $$aErin J. Adams 000005220 72014 $$aJustin P. Kline 000005220 72014 $$aHans Schreiber 000005220 8564_ $$93d6f8de4-7018-4d96-99f4-546a112f1b51$$s17434753$$uhttps://knowledge.uchicago.edu/record/5220/files/Hu_uchicago_0330D_16649.pdf$$ePublic 000005220 909CO $$ooai:uchicago.tind.io:5220$$pDissertations$$pGLOBAL_SET 000005220 983__ $$aDissertation