Investigation of oncogenic G‐protein coupled receptor signaling pathways in Kras dependent pancreatic cancer cell lines

P ancreatic D uctal A denocarcinoma (PDA) is the most common form of pancreatic cancer driven primarily by mutations in V ‐Ki‐Ras2 K irsten Ra t S arcoma Viral Oncogene Homolog ( Kras ) gene. Its most common mutation, G12D, cause KRAS to remain constitutively active. Kras G12D expressing pancreatic acinar cells undergo transdifferentiation into ductal phenotype and form early tumorigenic lesions called Pan creatic I ntraepithelial N eoplasm (PanIN). Coincidentally occurring inactivating mutations of tumor suppressor genes, such as C yclin‐ d ependent K inase I nhibitor 2A ( Cdkn2a ), T umor P rotein p 53 ( Tp53 ), and M others A gainst D ecapentaplegic Homolog 4 ( Smad4 ) trigger progression of PanINs through stages from 1A to 3, eventually leading to carcinoma in situ and metastasis. G ‐ p rotein C oupled R eceptors (GPCR) are transmembrane receptors that transfer extracellular signals to intracellular heterotrimeric G‐proteins with Gα and Gβγ subunits. Upon activation, dissociated Gα and Gβγ subunits separately induce intracellular second messengers for key cellular responses involved in various aspects of development, metabolism, and cell motility. GPCR signaling is negatively regulated by intracellular R egulator of G ‐protein S ignaling (RGS) proteins that deactivate G‐protein by mediating re‐association of Gα and Gβγ subunits. Although GPCRs are the largest non‐antibiotic drug targets, their specific role in pancreatic cancer is not much known. GPCR signaling can influence RTK pathways in cross‐talk via various second messengers. The interplay between these two major classes of cellular pathways employing both heterotrimeric and monomeric G‐proteins has therefore potential to shed new light on pancreatic cancer research. We have previously identified Rgs16 expression as a marker of pancreatic tumor formation in Rgs16::GFP o/o ; Ptf1a Cre/+ ; Lox‐STOP‐Lox‐Kras G12D/+ ; Cdkn2a f/f (Rgs16::GFP‐KIC) mice from the onset of earliest PanINs at two weeks of age (Dis. Model Mech. 8, 1201). Using ImageJ software to quantitatively determine eGFP expression in micrographs, we have developed a r apid i n v ivo a ssay (RIVA) where we represent pancreatic tumor burden by one month of age and compare outcomes of known and novel therapeutics following two week long drug regimens. We found that just like the recently developed drug, BGB324, targeting an important RTK, called A ne x e l ekto ( Axl ), the anticoagulant warfarin was more effective in reducing tumor burden than standard chemotherapy alone. Currently, my research focus is centered on understanding active GPCR pathways that stimulate cell growth, survival, and migration by enhancement of Kras downstream signaling. My aim is comparison of the effects of general G‐protein signaling inhibition in Kras mutant and wild‐type cell lines, such as AsPC‐1 in contrast to BxPC‐3, on cell behavior and making connections to active GPCR signaling components. Support or Funding Information Supported by NCI CA161624. This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .