Abstract 5042: CREB transgenic mice to study alcohol-associated pancreatic carcinogenesis
Authors: Totiger Tulasigeri , Srinivasan Supriya , VanSaun Michael , Joshi Chandrashekhar , Shi Chanjuan , Dai Xizi , Dawra Rajinder , Gaidarski Alexander , Nestler Eric , Merchant Nipun , Nagathihalli Nagaraj 
Published: August 2018

Background: Pancreatic ductal adenocarcinoma (PDAC) is the third leading cause of cancer-related deaths in the United States. Chronic alcohol (&gt;60 grams/day) consumption is strongly associated with the risk of PDAC development. The metabolites generated from alcohol have been shown to cause significant pancreatic injury. Although the association of alcohol with PDAC progression has been established, the details of the underlying molecular and cellular mechanisms governing this process are unknown. Our study revealed that alcohol-associated pancreatic carcinogenesis correlates with CXCL12-induced activation of cyclic AMP response element binding protein (CREB). In this study, we examined newly developed CREB knockout (CREBfl/fl) mouse in pancreas.

Methods: Human pancreatic epithelial lines (HPNE), HPNE cells with KRAS (HPNE-KRAS), pancreatic stellate cells (PSCs), pancreatic intraepithelial neoplasia (PanIN) mouse cell lines (LSL-KrasG12D/+; Pdx1Cre/+) and cancer-associated fibroblasts (CAFs) cells were exposed to chronic alcohol (50 mM) and analyzed for phospho-kinases in cell lysates and cytokines in conditioned media. Inducible Ptf1aCreERTM;KRASG12D/+ (iPK) mice and CREB knockout (CREBfl/fl) with KRAS activation [Ptf1aCreERTM;KRASG12D/+;CREBfl/fl (iPKCREBKO)] mice were used to investigate the effect of alcohol on CREB activation and the role of CREB in alcohol-associated PDAC pathogenesis. iPK and iPKCREBKO mice were exposed to Lieber-DeCarli alcohol diet for up to 14 weeks with or without caerulein injections. The number of acinar cells (amylase), ducts (cytokeratin 19), PanIN lesions (alcian blue positive), fibrosis (sirius red) and activation of CREB were measured by immunohistochemistry at 6 and 14 weeks of alcohol exposure in vivo. Serum obtained from alcohol-fed iPK and iPKCREBKO mice were analyzed for significant cytokine release upon alcohol exposure in vivo. We then determined the biologic effects of pharmacologic CREB inhibition in iPK mice exposed to alcohol diet.

Results: We found increased pCREB levels in HPNE and HPNE-KRAS cells upon treatment with alcohol, which was further associated with the up-regulation of the CXCL12 protein in the conditioned media. Serum CXCL12 and tissue pCREB levels were high in alcohol fed iPK mice when compared to their corresponding control diet-fed mice. Blocking CXCL12 with Plerixafor, a CXCL12-CXCR4 axis inhibitor, decreased alcohol-induced pCREB levels. Finally, exposure of iPK mice to an alcohol diet coupled with cerulein administration significantly increased the number of PanIN lesions (alcian blue+ cells), and decreased acinar cells when compared to alcohol-fed iPKCREBKO mice.

Conclusion: These findings implicate CREB as a critical oncogenic driver in alcohol addiction-induced pancreatic carcinogenesis. Additionally, CXCL12 represents an important mediator of CREB activation, which can be pharmacologically targeted.

Citation Format: Tulasigeri Totiger, Supriya Srinivasan, Michael VanSaun, Chandrashekhar Joshi, Chanjuan Shi, Xizi Dai, Rajinder Dawra, Alexander Gaidarski, Eric Nestler, Nipun Merchant, Nagaraj Nagathihalli. CREB transgenic mice to study alcohol-associated pancreatic carcinogenesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 5042.