LAUSR

Bispecific T cell engager (BiTE®) antibody constructs are designed to redirect T cells to induce lysis of tumor cells through simultaneous binding to CD3 on T cells and to a tumor associated antigen. BiTE® antibody constructs have previously been shown to be effective at depleting blood and tissue targets in B cell malignancies, suggesting a therapeutic potential for BiTE® antibody constructs in solid tumors. Pancreatic ductal adenocarcinoma (PDA) is a devastating malignant disease with a dismal prognosis even with currently available therapies. The tumor differentiation antigen mesothelin (MSLN) is highly expressed in over 80% of pancreatic tumors. Expression of MSLN is restricted in normal tissue implicating MSLN as an attractive target for BiTE® targeted therapy in PDA. In this study, we report the preclinical characterization of an anti-MSLN/CD3 BiTE® antibody construct with half-life extension (MSLN HLE BiTE®) in xenograft models of PDA. Previously the MSLN HLE BiTE® was found to bind CD3 and MSLN with low nanomolar affinity and have low picomolar cytotoxic activity against MSLN-positive cells in vitro. Moreover, the MSLN HLE BiTE® can mediate redirected lysis of cancer cell lines resistant to chemotherapy. Our study examines the efficacy of the highly potent and specific MSLN HLE BiTE® antibody construct in vivo by utilizing the orthotopic implantation of human AsPC-1 luciferase expressing PDA cells in immunocompromised NOD-Prkdcscid IL2rgnull (NSG) mice. We found that mice receiving the MSLN HLE BiTE® antibody construct survived significantly longer and had significantly reduced tumor burden comparing to control mice. Moreover, tumor bearing mice were given fluorescently labeled human T cells to measure T cell localization. Using live animal fluorescent imaging, we found mice treated with the MLSN HLE BiTE® antibody construct had significantly increased T cell localization and retention to the PDA tumor area compared to control. Mice receiving orthotopic implantation of MSLN knock out human AsPC-1 tumors treated with the MSLN HLE BiTE® did not show increased survival, reduction of tumor burden or increased T cell localization, confirming the BiTE® specificity for the tumor target MSLN. Furthermore, using the KPC mouse model of PDA, which develops cancer spontaneously upon conditional pancreatic expression of KRASG12D and TP53R172H mutations, we tested if a mouse-specific surrogate MSLN HLE BiTE® antibody construct could localize to a naturally formed dense tumor microenvironment. We found that mice treated with a murine MSLN HLE BiTE® labeled with CW800 demonstrated strong localization of the MSLN HLE BiTE® through the dense microenvironment to the PDA tumor. Taken together, these results suggest that a MSLN HLE BiTE® may be a unique targeted immunotherapy for PDA patients. Citation Format: Noelle R. Jurcak, MacKenzie Zarecki, Fei Lee, Noah Rozich, Stephen Muth, Elizabeth Jaffee, Sabine Stienen, Julie Bailis, Lei Zheng. Evaluation of mesothelin BiTE® antibody constructs in models of pancreatic ductal adenocarcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 1561.