Abstract B43: p120 catenin loss drives pancreatic cancer EMT and metastasis through activation of PTHrP-mediated calcium signaling

Abstract

Abstract Purpose: An unbiased approach to discover candidate cancer genes in pancreatic ductal adenocarcinoma (PDAC) identified the p120 catenin gene (Ctnnd1) as one of the top 20 driver genes, and further analysis revealed that P120CTN loss correlated with reduced survival. Results presented herein used genetically engineered mouse models (GEMMs) to show that P120CTN is a potent metastasis suppressor in PDAC. Methods: We employed orthotopic injection, tail vein metastasis assays, and mouse hospital preclinical trials to show that deletion or pharmacologic inhibition of Parathyroid Hormone Related Protein (PTHrP/PTHLH), a signaling element downstream of P120CTN, delayed tumor development and metastatic outgrowth. Finally, we generated a novel GEMM of Pthlh deletion to demonstrate in the autochthonous KPC model that loss of PTHrP phenocopies anti-PTHrP therapy by blocking both primary and metastatic tumor growth. Results: We have generated a mouse model to delete the gene Ctnnd1, whose gene product p120 catenin (P120CTN) is necessary for E-cadherin stability, resulting in enhanced epithelial-to-mesenchymal transition (EMT) and metastasis in KPC animals. Specifically, we show that KPC-p120ctncKO mice have a dramatically enhanced metastatic phenotype relative to KPC controls, suggesting that P120CTN is a critical factor in metastatic cell dissemination. An unbiased screen of tumor cells isolated from these mice identified misregulated calcium signaling through the Parathyroid Hormone Related Protein (PTHrP) as a previously unappreciated contributor to EMT and metastasis. Genetic deletion of the gene that codes for PTHrP in orthotopic transplantation experiments showed significantly reduced tumor growth and metastasis, establishing PTHrP as an oncogenic and prometastatic secreted peptide. Furthermore, treatment with anti-PTHrP monoclonal antibodies reduced tumor cell proliferation and migration in vitro, demonstrating that anti-PTHrP therapies may be of clinical benefit. Importantly, we generated KPC-PthlhcKO mice and showed that they have significantly reduced primary and metastatic tumor burden and increased survival relative to KPC controls. In parallel experiments, we treated KPC mice in a preclinical trial with anti-PTHrP neutralizing antibodies, which delayed both primary and metastatic tumor growth. Finally, analysis of human samples demonstrated that increased PTHLH expression is associated with significantly decreased survival, and that a subset of patients have PTHLH genomic amplifications. Conclusions: This novel work has demonstrated the importance of the previously unappreciated role that PTHrP-mediated calcium signaling plays in pancreatic cancer cellular plasticity and metastasis, and future studies will look to determine the efficacy of anti-PTHrP monoclonal antibodies with a view towards translation in human clinical trials. Citation Format: Jason R. Pitarresi, Robert J. Norgard, Ben Z. Stanger, Anil K. Rustgi. p120 catenin loss drives pancreatic cancer EMT and metastasis through activation of PTHrP-mediated calcium signaling [abstract]. In: Proceedings of the AACR Special Conference on Pancreatic Cancer: Advances in Science and Clinical Care; 2019 Sept 6-9; Boston, MA. Philadelphia (PA): AACR; Cancer Res 2019;79(24 Suppl):Abstract nr B43.