EXTH-33. COMBATING TUMOR HETEROGENEITY IN GLIOBLASTOMA WITH A NOVEL CAR-BASED PLATFORM

Abstract Tumor heterogeneity is a major hindrance impeding success of antigen targeting immunotherapies, such as chimeric antigen receptor (CAR) T cells, in solid tumors. CAR T cells engineered to target EGFRvIII, IL13Rα2, and HER2 have mostly failed clinically to treat glioblastoma, because these antigens are typically only expressed on a subset of a tumor cells within a patient. Thus, finding a surface target that more universally characterizes tumor cells will likely be critical if antigen-specific immunotherapies are to succeed in glioblastoma and other solid tumors. Recently, we uncovered a mechanism whereby CD8 T cells can directly kill tumor cells independently of tumor-expressed major histocompatibility complex (MHC)-I. We found this mechanism to be dependent on NKG2D expression on T cells and NKG2D ligand (NKG2DL) expression on tumor cells. Interestingly, NKG2DL expression was not limited to MHC-I-deficient tumor cells, nor was it limited to gliomas, but NKG2DL expression characterized a variety of solid tumor types. Therefore, NKG2DL is potentially an ideal target for immune-based therapy. Here, we developed a bi-specific fusion protein comprised of the extracellular domain of NKG2D linked to an anti-CD3scFv. Thus, T cells will be redirected to target NKG2DL-expressing tumor cells. We have termed this fusion protein Bi-specific Engagers without Antigen Requirements (BEAR). To ensure local, intratumoral delivery of BEAR, we engineered an EGFRvIII-targeting CAR T cell to secrete BEAR (VIII CAR Engineered with BEAR; VIII “CARE-BEAR”). VIII CARE-BEAR demonstrated potent tumor kill in both in vitro and in vivo tumor heterogeneity models. Moreover, we engineered an EGFRwt-targeting (D2C7) CARE-BEAR to exemplify the potential utility of CARE-BEAR beyond glioblastoma.