Abstract B63: Improving CAR T cell function by reversing the immunosuppressive tumor environment of pancreatic cancer

Background: Adoptive transfer of T cells redirected to tumor-associated antigens (TAAs) by expression of chimeric antigen receptors (CARs) can produce tumor responses, even in patients with resistant malignancies. Although these CAR-T cells have potent anti-tumor activity in vitro and in vivo, pancreatic tumors employ immune evasion mechanisms, such as the production of inhibitory cytokines, which limit in vivo CAR-T cell persistence and effector function. Methods: To target pancreatic ductal adenocarcinoma (PDAC), we generated T cells expressing a CAR directed to the TAA prostate stem cell antigen (PSCA). We also engineered a chimeric cytokine receptor in which the IL4 receptor exodomain was fused to the IL7 receptor endodomain (IL4/7 ChR). Expansion and selection profiles and short- and long-term anti-tumor activity of these transgenic T cells were assessed. Results: T cells expressing CAR-PSCA kill PSCA(+) tumor cell lines CAPAN1 and K562-PSCA but not PSCA(-) targets, such as 293T (74±4%, 73±6% and 9±3% specific lysis, respectively, 10:1 E:T, n=3). Although these CAR-T cells had potent anti-tumor activity in vitro and in vivo, pancreatic tumors employ immune evasion mechanisms, such as the production of inhibitory cytokines, which limit in vivo CAR-T cell persistence and effector function. Indeed, when the serum of patients with pancreatic cancer (n=8) was examined, we found the levels of the immunosuppressive cytokine IL4 to be elevated relative to patients with benign pancreatic disorders or normal healthy controls (14.25±19.48 pg/mL vs 7.28±9.03 vs 1.13±1.42 pg/mL). Thus, to protect the CAR-PSCA T cells from the negative influences of IL-4, we generated a chimeric cytokine receptor in which the IL4 receptor exodomain was fused to the IL7 receptor endodomain (IL4/7 ChR). Transgenic expression of this molecule in CAR-PSCA T cells should invert the inhibitory effects of tumor-derived IL4 and instead promote the proliferation of the effector CAR T cells. In preliminary experiments we successfully co-expressed both CAR-PSCA and IL4/7 ChR (47.5±12.3% double-positive cells, n=4) on primary T cells. These T cells retained their tumor-specific activity (80±8% specific lysis against CAPAN1, 10:1 E:T, n=3) and when cultured in conditions that mimic the tumor milieu (IL4 12.5 ng/ml), CAR-PSCA 4/7R ChR-modified T cells continued to expand unlike unmodified CAR-PSCA T cells (from 2x106 cells on day 0 to 5.53x1010±8.46x1010 cells (CAR-PSCA 4/7R ChR) on day 28, in comparison to CAR-PSCA T cells that reached only 3.84x108±5.43x108 cells, n=4). Indeed, in the presence of IL4, transgenic cells had a selective advantage (comprising 44.8±11.0% of the population on day 0 and 87.6±10.0% on day 28, n=4), but even after prolonged cytokine exposure these T cells remained both antigen- and cytokine-dependent. Conclusions: CAR-PSCA 4/7 ChR-modified tumor-specific T cells can effectively target pancreatic cancer cells and should be equipped to expand, persist, and retain their cytotoxic function even in the presence of high levels of IL4 in the tumor microenvironment. Citation Format: Somala Mohammed, Sujita Sukumaran, Usanarat Anurathapan, Pradip Bajgain, Helen E. Heslop, Cliona M. Rooney, Malcolm K. Brenner, Ann M. Leen, Juan F. Vera. Improving CAR T cell function by reversing the immunosuppressive tumor environment of pancreatic cancer. [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Innovations in Research and Treatment; May 18-21, 2014; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2015;75(13 Suppl):Abstract nr B63.