Engineering best in class Acute Myeloid Leukemia (AML) CAR-T therapies by enhancing persistence and defining the immune landscape of AML - PROJECT SUMMARY/ABSTRACT Acute Myeloid Leukemia (AML) is a devastating disease with <30% 5-year survival on current treatment regimens. This cancer, diagnosed in >20,000 Americans per year, has an urgent need for new therapeutic strategies. One of the most exciting new treatment strategies for blood cancers are Chimeric Antigen Receptor (CAR) T cells, which have led to cures in patients with B-cell origin malignancies who previously had dismal prognoses. However, current implementations of CAR-T cells have been much less successful in AML. Translating CAR-T cells to AML appears to be hampered by 3 major hurdles: 1) lack of a highly disease specific antigen due to immense heterogeneity of the AML tumor microenvironment (TME) 2) elevated on target-off tumor toxicities where potential therapeutic targets are also expressed on healthy hematopoietic blood cells 3) and the presence of AML TME factors that impede T-cell persistence and lead to antigen positive relapse. My long term goal in this proposal is to develop a cellular therapy platform to overcome these hurdles in current AML CAR-T therapies. This proposal will be led by myself under the sponsorship of Dr. Arun Wiita, an expert in hematologic malignancies and cellular therapy development as well as a team of collaborators with complementary and relevant expertise. This work builds on two key recent discoveries: 1) data recently published by the Wiita lab (Mandal et al, Nature Cancer 2023) where our group discovered the active conformation of Integrin Beta-2 (aITGB2) as a novel and selective CAR-T target for AML, and 2) the discovery by the Jaehyuk Choi lab at Northwestern, my close collaborator on this proposal, of the CARD11-PIK3R3 fusion gene as one of the most potent genetic modifications to T-cells to drive long-term cellular therapy persistence in vivo (Garcia and Daniels et al, Nature, in press). Here I hypothesize that by combining these two technologies, I can develop a best in class AML therapy that overcomes the previous hurdles described. My major experimental goals include 1) to develop engineered T-cells that localize to AML blasts (in vitro/in vivo) and enhance CAR-T therapy with minimal toxicities and high persistence and 2) to quantify and identify the molecular changes that are accrued in tumor cells and neighboring immune cells in response to CAR-T therapy. In Aim 1 I will test the feasibility, toxicity, efficacy, and persistence of the CARD11-PIK3R3 fusion enhanced, AML specific, CAR-T in-vitro and in-vivo (PDX and Syngeneic mouse models). In Aim 2 I aim to identify molecular changes by single cell RNA sequencing that are accrued in response to our new CAR-T therapy at initial treatment, peak response, and relapse in-vivo within CAR-T, neighboring immune cell, and tumor cell populations. Successful completion of these aims will enable us to enhance persistence of current therapies against existing targets in AML. Completion of this work in conjunction with the associated training plan will enable me the support to become an independent scientist focused on developing new treatment modalities. Furthermore, my graduate training will be tailored to me along the path of becoming an independent investigator with a lab that integrates bioengineering and immunology.
