Pancreatic ductal adenocarcinoma (PDAC) is one of the deadliest forms of cancer. Poor survival rates are largely due to the late stage at which PDAC is diagnosed and a lack of effective therapies. The long-term goal of this research program is to discover new immunotherapeutic approaches for the treatment of PDAC. Preliminary studies by our group and others have shown that the Anterior Gradient-2 (AGR2) protein, a member of the protein disulfide isomerase (PDI) family, is induced during PDAC oncogenesis and highly expressed in >90% of PDAC patients. AGR2 has intracellular oxidative folding function and is also released from the cell where it localizes to the surface of PDAC cells and is shed into the tumor microenvironment. We hypothesize that AGR2 is an actionable target for the development of PDAC targeted immunotherapies and will test that theory using a new immunotherapy drug candidate. In preliminary studies we generated a proprietary anti-AGR2 single chain antibody fragment (scFv) by phage display screening and constructed a bispecific T-cell engager (BiTE) that binds to AGR2 and CD3, which is a subunit of the T cell receptor complex. We found that BiTE engagement stimulates T cell receptor signaling, T cell activation/proliferation, and T cell induced killing of AGR2-positive PDAC cells in cellular models. The specific objectives of this study are: (1) to improve the AGR2 binding affinity of our scFv via affinity maturation, and (2) to demonstrate high potency T cell dependent killing of PDAC cells by an optimized BiTE molecule in vitro and in genetically engineered mouse models (GEMMs) of PDAC. These aims are built on clear rationale from the existing literature and strong preliminary data. This work is innovative because we will investigate the activity and mechanism of a new immunotherapy drug candidate for the treatment of PDAC, which is a cancer in need of new therapies. In addition we expect that this study will reveal new methods and mechanisms for generating an anti-tumor immune response in PDAC, for which existing immunotherapies have thus far been ineffective. Public Health Relevance Statement Narrative (Relevance)) ---------------------------- Pancreatic cancer is one of the deadliest cancers and effective therapies are lacking. We have developed an immunotherapy approach to target the Anterior Gradient-2 (AGR2) protein, which is highly expressed in pancreatic tumors. Our strategy relies on a Bispecific T-cell Engager (BiTE) that activates immune cells in pancreatic tumors and causes an anti-tumor immune response. We anticipate that these studies will reveal new ways of treating pancreatic cancer in preclinical models of cancer, setting the stage for clinical development and the delivery of a new treatment regimen to patients in need.
Project Terms: Bacteriophages ; Phages ; bacterial virus ; Biological Assay ; Assay ; Bioassay ; Biologic Assays ; Blood ; Blood Reticuloendothelial System ; Malignant Neoplasms ; Cancers ; Malignant Tumor ; malignancy ; neoplasm/cancer ; Cell Culture Techniques ; cell culture ; Cells ; Cell Body ; Diagnosis ; Enzyme-Linked Immunosorbent Assay ; ELISA ; Family ; Goals ; Human ; Modern Man ; Immunoglobulin Fragments ; Antibody Fragments ; Complementarity Determining Regions ; Complimentarity Determining Region ; Hypervariable Loop ; Hypervariable Regions ; Immunoglobulin Hypervariable Region ; Immunotherapy ; Immune mediated therapy ; Immunologically Directed Therapy ; immune therapeutic approach ; immune therapeutic interventions ; immune therapeutic regimens ; immune therapeutic strategy ; immune therapy ; immune-based therapies ; immune-based treatments ; immuno therapy ; In Vitro ; Interferon Type II ; Gamma interferon ; IFN-Gamma ; IFN-g ; IFN-γ ; IFNG ; IFNγ ; Immune Interferon ; Interferon Gamma ; Interferon-gamma ; lFN-Gamma ; Lead ; Pb element ; heavy metal Pb ; heavy metal lead ; Libraries ; Literature ; Methods ; Mus ; Mice ; Mice Mammals ; Murine ; Mutation ; Genetic Alteration ; Genetic Change ; Genetic defect ; genome mutation ; Pancreas ; Pancreatic ; pancreatic neoplasm ; Pancreas Neoplasms ; Pancreas Tumor ; Pancreatic Tumor ; pancreatic neoplasia ; Parents ; Patients ; Proteins ; T-Cell Receptor ; MHC Receptor ; Major Histocompatibility Complex Receptor ; Receptors, Antigen, T-Cell ; Research ; Specificity ; Survival Rate ; T-Lymphocyte ; T-Cells ; thymus derived lymphocyte ; Testing ; Thermodynamics ; Thermodynamic ; Tissues ; Body Tissues ; Translations ; Treatment Protocols ; Treatment Regimen ; Treatment Schedule ; Work ; Protein Disulfide Isomerase ; Disulfide Interchange Enzyme ; Disulfide Isomerase ; ERp59 PDI ; Erp59 ; GSBP ; Glycosylation Site-Binding Protein ; S-S rearrangase ; Sulfhydryl-Disulfide Interchange Enzyme ; Thiol-Disulfide Transhydrogenase ; Trypanothione-Glutathione Thioltransferase ; dsbA Gene Product ; dsbA Protein ; dsbC Gene Product ; dsbD Gene Product ; xprA Gene Product ; Measures ; Oligonucleotide-Directed Mutagenesis ; Muromonab-CD3 ; Anti-CD3 Monoclonal Antibody OKT3 ; MoAb OKT3 ; Monoclonal Antibody OKT3 ; Muromonab-CD3 Monoclonal Antibody ; OKT3 ; CD3 Antigens ; CD3 ; CD3 Complex ; CD3 molecule ; OKT3 antigen ; T3 Antigens ; T3 Complex ; T3 molecule ; protein folding ; improved ; Anterior ; Surface ; Phase ; Collaborations ; Cell-Mediated Lympholysis ; Cellular Cytotoxicity ; Lymphocyte Cytotoxicity ; Lymphocytotoxicity ; cell mediated cytotoxicity ; Cell-Mediated Cytolysis ; Therapeutic ; fluid ; liquid ; Liquid substance ; Malignant Pancreatic Neoplasm ; Pancreas Cancer ; Pancreatic Cancer ; pancreatic malignancy ; Malignant neoplasm of pancreas ; programs ; Immunes ; Immune ; Complex ; Dependence ; subdermal ; subcutaneous ; extracellular ; mutant ; Tumor Cell ; neoplastic cell ; Surface Plasmon Resonance ; established cell line ; tumor growth ; cell killing ; member ; Position ; Positioning Attribute ; (TNF)-α ; Cachectin ; Macrophage-Derived TNF ; Monocyte-Derived TNF ; TNF ; TNF A ; TNF Alpha ; TNF-α ; TNFA ; TNFα ; Tumor Necrosis Factor ; Tumor Necrosis Factor-alpha ; TNF gene ; Modeling ; Sampling ; theories ; immune drugs ; immune-based therapeutics ; immunologic preparation ; immunologic therapeutics ; immunotherapeutics ; immunotherapy agent ; Immunotherapeutic agent ; Cancer Treatment ; Malignant Neoplasm Therapy ; Malignant Neoplasm Treatment ; anti-cancer therapy ; anticancer therapy ; cancer-directed therapy ; cancer therapy ; model development ; T-Cell Activation ; Molecular Interaction ; Binding ; Oncogenesis ; tumorigenesis ; Pancreas Ductal Adenocarcinoma ; Pancreatic Ductal Adenocarcinoma ; Affinity ; Data ; Preclinical Models ; Pre-Clinical Model ; Receptor Signaling ; in vivo ; Cancer Model ; CancerModel ; Phage Display ; Tissue Microarray ; Tissue Arrays ; Tissue Chip ; Validation ; Xenograft Model ; xenograft transplant model ; xenotransplant model ; Immunologics ; Immunochemical Immunologic ; Immunologic ; Immunological ; Immunologically ; Genetically Engineered Mouse ; GEM model ; genetically engineered mouse model ; genetically engineered murine model ; pancreas development ; Development ; developmental ; pre-clinical ; preclinical ; tumor microenvironment ; cancer microenvironment ; knock-down ; knockdown ; innovation ; innovate ; innovative ; Cell model ; Cellular model ; novel therapeutics ; new drug treatments ; new drugs ; new therapeutics ; new therapy ; next generation therapeutics ; novel drug treatments ; novel drugs ; novel therapy ; mouse model ; murine model ; commercialization ; tumor ; overexpression ; overexpress ; treatment strategy ; effective therapy ; effective treatment ; drug candidate ; screening ; targeted treatment ; targeted drug therapy ; targeted drug treatments ; targeted therapeutic ; targeted therapeutic agents ; targeted therapy ; humanized mouse ; humanized mice ; clinical candidate ; experimental study ; experiment ; experimental research ; clinical development ; immune checkpoint blockade ; check point blockade ; checkpoint blockade ; immune check point blockade ; anti-tumor immune response ; antitumor immune response ; anti-PD-1 ; aPD-1 ; aPD1 ; anti programmed cell death 1 ; anti-PD1 ; anti-programmed cell death protein 1 ; antiPD-1 ; antiPD1 ; αPD-1 ; αPD1 ; bi-specific T cell engager ; bispecific T cell engager ; in vivo evaluation ; in vivo testing ; pancreatic ductal adenocarcinoma cell ; PDAC cancer cell ; PDAC cell ; pancreatic ductal adenocarcinoma model ; PDA model ; PDAC Model ; KRASG12D ; KRAS G12D ;
