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Cigarette smoke extract inhibits fluid and HCO3− secretion and CFTR activity in guinea pig pancreatic ductal cells
S22
Abstracts / Pancreatology 16 (2016) S1eS130
seems advisable and future studies should identify risk factors and early treatment strategies for NOPD.
Abstract ID: 1547. Genome-wide RNAi screening identified metastasis suppressor genes in an orthotopic pancreatic cancer mouse model Yangchao Chen The Chinese University of Hong Kong, Hong Kong S.A.R., China Introduction: Pancreatic cancer is an aggressive malignancy with extremely poor prognosis. It is usuallly diagnosed when metastases are already present. Aims: To identify genes that play critical roles in the process of pancreatic cancer metastasis, a whole genome RNAi screening was performed. Materials & methods: An shRNA library targeting all human genes was introduced into a human pancreatic cancer cell line Capan-2. The infected cells were then transplanted into the pancreas of nude mice. Because Capan-2 is of low metastatic potential, we hypothesized that knocking down of metastasis suppressor genes would facilitate Capan-2 cells to spread to the liver. By retrieving shRNA templates from the liver metastatic nodules, several candidate genes were found. One fo them Sox9 has been validated as a metastasis suppressor gene in vivo, implying that loss of expression of Sox9 promoted pancreatic cnacer metastasis. Results: In the validation experiments, 10 and 11 nude mice were used for control shCTRL and shSox9 group respectively. All of them had primary tumors. 7 out of 11 mice in shSox9 group developed liver metastasis, whereas only 2 out of 10 metastasized in the control group. Knocking down Sox9 promoted the orthotopic transplanted Capan-2 cells to metastasize to the liver. Conclusion: Our genome wide RNAi screening identified Sox9 as a metastasis suppressor gene in pancreatic cancer.
Abstract ID: 1549. The utility of minichromosome maintenance proteins as novel diagnostic markers in pancreaticobiliary malignancy Margaret G. Keane 1, Matthew T. Huggett 1, Tu Vinh Luong 2, Douglas Thorburn 1, Gavin J. Johnson 3, Michael H. Chapman 3, George J. Webster 3, James Mackay 4, Gareth Williams 5, Stephen P. Pereira 1 1 University College London, Institute for Liver and Digestive Health, United Kingdom 2 Department of Cellular Pathology, Royal Free London NHS Foundation Trust, United Kingdom 3 University College London, NHS Foundation Trust, United Kingdom 4 University College London, Department of Genetics Evolution and Environment, United Kingdom 5 University College London, Wolfson Institute for Biomedical Research, United Kingdom
Introduction: Pancreatic cancer is the fourth commonest cause of cancer death, with a 5-year survival of <4%. Earlier diagnosis and detection of precursor lesions with significant malignant potential would enable timely surgical resection. Minichromosome maintenance proteins (Mcm 2e7) are expressed by actively cycling malignant tumours. Aims: Evaluate if Mcm protein expression can differentiate premalignant and malignant pancreaticobiliary disease. Materials & methods: 1.) Mcm2 protein expression was assessed in formalin-fixed paraffin embedded (FFPE) tissue samples. Sections were immunostained and protein expression analysis performed by determining the labelling index (positive cells / total number of cells). 2.) An automated immunocolormetric assay was used to measure levels of Mcm5 in: (i) 97 biliary brush (BB) samples obtained at endoscopic retrograde cholangiopancreatography and (ii) 28 pancreatic cyst fluid (PCF) samples obtained during endoscopic ultrasound.
Results: FFPE tissue samples from 73 patients with pancreatic cancer, 44 patients with a range of cystic tumours and 9 patients with benign pancreatic disease (normal pancreas or chronic pancreatitis) were stained for Mcm2. Mcm2 protein expression in pancreatic cancer, mucinous cysts, serous cystadenomas and benign disease was 62.2%, 35.5%, 9.3% and 0.4% respectively (p<0.05). Mcm5 levels in BB samples were more sensitive than brush cytology for the detection of malignancy (62% vs. 47%; p<0.05) in patients with an indeterminate biliary stricture. In pancreatic cysts Mcm5 detected malignancy with a sensitivity of 50% (specificity 73%) and measurement was feasible in small and acellular samples. Conclusion: Mcm protein expression can differentiate malignant, premalignant and benign pancreaticobiliary diseases. For BB samples, the Mcm5 assay was superior to cytology.
Abstract ID: 1551, Oral-1. Pancreatic ductal fluid- and HCO3-secretion is reduced in aquaporin 1 and 4 knock-out mice zi 1, Petra Pallagi 2, Anita Bala zs 2, Matthias Sendler 3, Jens Eszter Becskeha n Rakonczay, Jr. 2, P ria Venglovecz 1 eter Hegyi 4, Vikto Kühn 3, Zolta 1 Department of Pharmacology and Pharmacotherapy, University of Szeged, Hungary 2 First Department of Medicine, University of Szeged, Hungary 3 Ernst-Moritz-Arndt-University, Greifswald, Germany 4 cs, Institute for Translational Medicine & Department University of Pe of Translational Medicine/1st Department of Medicine, MTA-SZTE Lendület Translational Gastroenterology Research Group, University of Szeged, Hungary
Introduction: Acute pancreatitis (AP) is a multicellular disease which usually associated with impaired fluid secretion by the ductal cells. Contribution of aquaporins (AQPs) to this fluid secretion is less characterized. Aims: our aim in this study was to determine the role of AQPs in ductal fluid and HCO-3 secretion using AQP knock out (KO) mice. Materials & methods: Intra/interlobular pancreatic ducts were isolated from mice by collagenase digestion. Pancreatic HCO-3 secretion was measured using the Cl- withdrawal technique, whereas the rate of ductal fluid secretion was determined by video microscopy. Pancreatic juice secretion was examined in vivo in anesthetized mice using magnetic resonance imaging cholangiopancreatography. Results: Intracellular pH alkalinisation induced by removal of luminal Cl- was significantly reduced in AQP-1 (42±3,2%) and AQP-4 (52±4,5%) KO vs. wild type (WT) mice. There were no significant changes in the basal fluid secretion between WT and KO ducts. In contrast, stimulation of the ducts with 5 mM forskolin caused swelling of the WT ducts, whereas ducts from AQP-1 and AQP-4 KO mice showed no or only a slight response to forskolin. On retro-orbital injection of 10 U/kg body weight secretin, the increase in total excreted volume (TEV) in WT animals (0.023 TEV/cm3) was significantly higher than in AQP-1 (0.0041 TEV/ cm3) and AQP-4 KO animals (0.0068 TEV/ cm3). Conclusion: Pancreatic fluid and HCO-3 secretion secretion significantly reduced in AQP-1 and AQP-4 KO mice, indicating that AQPs play an essential role in pancreatic fluid secretion. Supported by OTKA (NF105758, NF100677, K109756) and MTA-SZTE Momentum Grant (LP2014-10/2014)
Abstract ID: 1553. Cigarette smoke extract inhibits fluid and HCO3 activity in guinea pig pancreatic ductal cells
secretion and CFTR
ria Venglovecz 2, Krisztina To th 1, Andrea Schnúr 1, Petra Pallagi 1, Vikto n Rakonczay, Jr. 4, Kata zsef Mal } Csupor 3, Zolta th 1, Jo eth 1, Dezso Emese To 5 5 6 } Cseko , Zsuzsanna Helyes , Peter Hegyi
Abstracts / Pancreatology 16 (2016) S1eS130 1
University of Szeged, First Department of Medicine, Hungary University of Szeged, Department of Pharmacology and Pharmacotherapy, Hungary 3 University of Szeged, Department of Pharmacognosy, Hungary 4 University of Szeged, First Department of Medicine, Department of Pathophysiology, Hungary 5 cs, Department of Pharmacology and University of Pe Pharmacotherapy, Hungary 6 cs, Institute for Translational Medicine & Department University of Pe of Translational Medicine/1st Department of Medicine, MTA-SZTE Lendület Translational Gastroenterology Research Group, University of Szeged, Hungary 2
Introduction: Smoking represents an independent risk factor for the development of chronic pancreatitis (CP). It is well documented that secretion of pancreatic ductal alkaline fluid (which is regulated mostly by anion exchangers and CFTR) is diminished in CP. Aims: In this study, we would like to understand whether smoking has any effects on pancreatic ductal fluid and HCO-3 secretion. Materials & methods: Guinea pigs were exposed to cigarette smoke four times a day for 30 min for 6 weeks. The expression of CFTR was analysed by immunohistochemistry. Intra/interlobular pancreatic ducts were isolated from guinea pig pancreas. Cigarette smoke extract (CSE) was prepared by smoking of 15 cigarettes into 10 ml distilled water by a smoking machine. Three different concentraion (20, 40 and 80mg/ml) were diluted using the stock solution. Intracellular pH was evaluated by microfluorometry. Basal and forskolin-stimulated fluid secretion was measured by video microscopy. CFTR currents were detected by whole cell configuration of patch clamp technique. Results: Cigarette smoking significantly diminished the expression of CFTR and the fluid and HCO-3 secretion in guinea pig pancreas. 40mg/ml CSE decreased HCO-3 secretion via inhibition of Cl-/HCO-3 exchanger activity. CSE dose-dependently decreased forskolin-stimulated fluid secretion in guinea pig pancreatic ducts and forskolin-stimulated Cl- current of CFTR Clchannel (20mg/ml by 44.5 %, 40mg/ml by 69.3 % and 80mg/ml by 81.3%). Conclusion: Smoking represents an independent risk factor for the development of chronic pancreatitis (CP). It is well documented that secretion of pancreatic ductal alkaline fluid (which is regulated mostly by anion exchangers and CFTR) is diminished in CP.
Abstract ID: 1554. Early life exposure to stressors promotes pancreatic carcinogenesis in KC mice Marl ene Dufresne 1, Sarah Gandarillas 1, Hubert Lulka 1, Pierre Dubus 2, Muriel Darnaud ery 3, Anne Gabory 4, Claudine Junien 4, Pierre Cordelier 1, ^me Torrisani 1 ero Michelle Kelly-Irving 5, Cyrille Delpierre 5, J 1
Inserm U1037, Cancer Research Center of Toulouse, France Bordeaux 2, EA2406, France Universite 3 gre e, France INRA UMR1286, Nutrition et Neurologie Inte 4 veloppement et Reproduction, UMR-INRA-ENVA-1198, Biologie du De France 5 Inserm, U1027, France 2
Introduction: Stressful experiences across the lifespan contribute to a variety of disorders including anxiety but also diseases like metabolic syndrome and cancer. Importantly, epidemiological studies suggested that cancer risk can be influenced by exposure to stressful conditions and events early on in life. Aims: To study the kinetic of PDAC development in KC mice that experienced one or two early life stressors by combining or not preconceptional nutritional stress and maternal separation. Materials & methods: One group of Pdx1-Cre females were fed with a control diet (CD) for 4 months and a second group received a high fat diet (HFD) for 2 months to induce obesity and then the CD for 2 months to induce weight loss. Females were then bred to LSL-KrasG12D+/- males to generate F1 KC. In maternal separation stressed groups, mouse pups were separated from their mother 3 hours per day from day 2 after birth and to
S23
day 14. Four groups of F1 KC were formed: from HFD mother ± separation, from CD mother ± separation. The pancreas phenotype was analyzed at 8 months of age. Results: Pre-conceptional HFD and/or maternal separation increased the density of preneoplastic lesions in the pancreas of F1 KC mice. We quantified more high-grade PanIN lesions and larger acinar-to-ductal metaplasia in the pancreas of stressed KC mice. Pre-conceptional HFD increased the size of low-grade PanIN and this of high-grade PanIN only when combined to maternal separation. Conclusion: Preconception maternal obesity and/or early life maternal separation promotes pancreatic carcinogenesis in KC mice.
Abstract ID: 1557. Serotonin-selective reuptake inhibitors (SSRI) as a novel approach to counteract pancreatic ductal adenocarcinoma progression Enrica Saponara 1, Gitta Seleznik 1, Raphael Buzzi 1, Irene Esposito 2, Johanna Bushmann 3, Francesco Baschieri 4, Hesso Farhan 5, Michele Visentin 6, Rolf Graf 1, Sabrina Sonda 1 1
Swiss HPB Center, Visceral & Transplantation Surgery, University Hospital Zurich, Switzerland 2 Institut für Pathologie, Universit€ atsklinikum Düsseldorf, Germany 3 Division of Reconstructive Surgery, Switzerland 4 rologie Gustave Roussy, Villejuif, France Institut de Cance 5 Department of Biology, University of Konstanz, Biotechnology Institute Thurgau, Switzerland 6 Department of Clinical Pharmacology and Toxicology, University Hospital Zurich, Switzerland Introduction: Pancreatic ductal adenocarcinoma (PDAC) is classified as one of the most recalcitrant amongst human malignancies due to its remarkable drug resistance. Nevertheless, little is known regarding the critical processes orchestrating PDAC progression, i.e. pancreatic intraepithelial neoplasia, accumulation of a dense stroma, angiogenesis and inflammatory environment. Aims: We previously demonstrated the role of serotonin (5-HT) in pancreatic inflammation and fibrotic processes during pancreatitis. We now aim to investigate the role of 5-HT in cancer progression and to propose potential new pharmacological strategies to counteract PDAC progression. Materials & methods: Transgenic mice harboring KrasG12D mutation, highly invasive human pancreatic cancer cell line (Panc1), human fibroblasts (HFF) and chick chorioallantoic membrane assay (CAM) were utilized to test the therapeutic potential of serotonin-selective reuptake inhibitors (SSRI). Transcriptome sequencing and imaging assays were used to characterize the effect of SSRI on Panc1 and HFF cell line. Results: Prolonged pharmacological treatment of KrasG12D mice with SSRI significantly reduced pre-malignant acinar cell transformation, along with a blunted desmoplastic stroma and inflammatory cell infiltration. Transcriptome analysis of Panc1 cells revealed that SSRI strongly interfered with cell cycle progression and microtubule assembly, concomitant with increased autophagy. Similar features were observed in SSRI-treated HFF cells displaying concomitant impaired cell motility. Finally, SSRI significantly reduced vessel number and diameter of highly vascularized CAM surfaces. Conclusion: Our data suggest that FDA-approved SSRI display alternative and unexpected clinical implications targeting both pre-malignant lesions and tumor micro-environment. Therefore, SSRI, in combination with chemotherapeutics, may constitute a novel approach to counteract PDAC progression.
Abstract ID: 1559. Characterization of pancreatic ductal fluid and bicarbonate secretion in wild type ferrets n th 1, Petra Pallagi 1, Jo zsef Mal ria Venglovecz 2, Zolta Emese To eth 1, Vikto 3 4 Rakonczay, Jr. , Peter Hegyi
Abstracts / Pancreatology 16 (2016) S1eS130
seems advisable and future studies should identify risk factors and early treatment strategies for NOPD.
Abstract ID: 1547. Genome-wide RNAi screening identified metastasis suppressor genes in an orthotopic pancreatic cancer mouse model Yangchao Chen The Chinese University of Hong Kong, Hong Kong S.A.R., China Introduction: Pancreatic cancer is an aggressive malignancy with extremely poor prognosis. It is usuallly diagnosed when metastases are already present. Aims: To identify genes that play critical roles in the process of pancreatic cancer metastasis, a whole genome RNAi screening was performed. Materials & methods: An shRNA library targeting all human genes was introduced into a human pancreatic cancer cell line Capan-2. The infected cells were then transplanted into the pancreas of nude mice. Because Capan-2 is of low metastatic potential, we hypothesized that knocking down of metastasis suppressor genes would facilitate Capan-2 cells to spread to the liver. By retrieving shRNA templates from the liver metastatic nodules, several candidate genes were found. One fo them Sox9 has been validated as a metastasis suppressor gene in vivo, implying that loss of expression of Sox9 promoted pancreatic cnacer metastasis. Results: In the validation experiments, 10 and 11 nude mice were used for control shCTRL and shSox9 group respectively. All of them had primary tumors. 7 out of 11 mice in shSox9 group developed liver metastasis, whereas only 2 out of 10 metastasized in the control group. Knocking down Sox9 promoted the orthotopic transplanted Capan-2 cells to metastasize to the liver. Conclusion: Our genome wide RNAi screening identified Sox9 as a metastasis suppressor gene in pancreatic cancer.
Abstract ID: 1549. The utility of minichromosome maintenance proteins as novel diagnostic markers in pancreaticobiliary malignancy Margaret G. Keane 1, Matthew T. Huggett 1, Tu Vinh Luong 2, Douglas Thorburn 1, Gavin J. Johnson 3, Michael H. Chapman 3, George J. Webster 3, James Mackay 4, Gareth Williams 5, Stephen P. Pereira 1 1 University College London, Institute for Liver and Digestive Health, United Kingdom 2 Department of Cellular Pathology, Royal Free London NHS Foundation Trust, United Kingdom 3 University College London, NHS Foundation Trust, United Kingdom 4 University College London, Department of Genetics Evolution and Environment, United Kingdom 5 University College London, Wolfson Institute for Biomedical Research, United Kingdom
Introduction: Pancreatic cancer is the fourth commonest cause of cancer death, with a 5-year survival of <4%. Earlier diagnosis and detection of precursor lesions with significant malignant potential would enable timely surgical resection. Minichromosome maintenance proteins (Mcm 2e7) are expressed by actively cycling malignant tumours. Aims: Evaluate if Mcm protein expression can differentiate premalignant and malignant pancreaticobiliary disease. Materials & methods: 1.) Mcm2 protein expression was assessed in formalin-fixed paraffin embedded (FFPE) tissue samples. Sections were immunostained and protein expression analysis performed by determining the labelling index (positive cells / total number of cells). 2.) An automated immunocolormetric assay was used to measure levels of Mcm5 in: (i) 97 biliary brush (BB) samples obtained at endoscopic retrograde cholangiopancreatography and (ii) 28 pancreatic cyst fluid (PCF) samples obtained during endoscopic ultrasound.
Results: FFPE tissue samples from 73 patients with pancreatic cancer, 44 patients with a range of cystic tumours and 9 patients with benign pancreatic disease (normal pancreas or chronic pancreatitis) were stained for Mcm2. Mcm2 protein expression in pancreatic cancer, mucinous cysts, serous cystadenomas and benign disease was 62.2%, 35.5%, 9.3% and 0.4% respectively (p<0.05). Mcm5 levels in BB samples were more sensitive than brush cytology for the detection of malignancy (62% vs. 47%; p<0.05) in patients with an indeterminate biliary stricture. In pancreatic cysts Mcm5 detected malignancy with a sensitivity of 50% (specificity 73%) and measurement was feasible in small and acellular samples. Conclusion: Mcm protein expression can differentiate malignant, premalignant and benign pancreaticobiliary diseases. For BB samples, the Mcm5 assay was superior to cytology.
Abstract ID: 1551, Oral-1. Pancreatic ductal fluid- and HCO3-secretion is reduced in aquaporin 1 and 4 knock-out mice zi 1, Petra Pallagi 2, Anita Bala zs 2, Matthias Sendler 3, Jens Eszter Becskeha n Rakonczay, Jr. 2, P ria Venglovecz 1 eter Hegyi 4, Vikto Kühn 3, Zolta 1 Department of Pharmacology and Pharmacotherapy, University of Szeged, Hungary 2 First Department of Medicine, University of Szeged, Hungary 3 Ernst-Moritz-Arndt-University, Greifswald, Germany 4 cs, Institute for Translational Medicine & Department University of Pe of Translational Medicine/1st Department of Medicine, MTA-SZTE Lendület Translational Gastroenterology Research Group, University of Szeged, Hungary
Introduction: Acute pancreatitis (AP) is a multicellular disease which usually associated with impaired fluid secretion by the ductal cells. Contribution of aquaporins (AQPs) to this fluid secretion is less characterized. Aims: our aim in this study was to determine the role of AQPs in ductal fluid and HCO-3 secretion using AQP knock out (KO) mice. Materials & methods: Intra/interlobular pancreatic ducts were isolated from mice by collagenase digestion. Pancreatic HCO-3 secretion was measured using the Cl- withdrawal technique, whereas the rate of ductal fluid secretion was determined by video microscopy. Pancreatic juice secretion was examined in vivo in anesthetized mice using magnetic resonance imaging cholangiopancreatography. Results: Intracellular pH alkalinisation induced by removal of luminal Cl- was significantly reduced in AQP-1 (42±3,2%) and AQP-4 (52±4,5%) KO vs. wild type (WT) mice. There were no significant changes in the basal fluid secretion between WT and KO ducts. In contrast, stimulation of the ducts with 5 mM forskolin caused swelling of the WT ducts, whereas ducts from AQP-1 and AQP-4 KO mice showed no or only a slight response to forskolin. On retro-orbital injection of 10 U/kg body weight secretin, the increase in total excreted volume (TEV) in WT animals (0.023 TEV/cm3) was significantly higher than in AQP-1 (0.0041 TEV/ cm3) and AQP-4 KO animals (0.0068 TEV/ cm3). Conclusion: Pancreatic fluid and HCO-3 secretion secretion significantly reduced in AQP-1 and AQP-4 KO mice, indicating that AQPs play an essential role in pancreatic fluid secretion. Supported by OTKA (NF105758, NF100677, K109756) and MTA-SZTE Momentum Grant (LP2014-10/2014)
Abstract ID: 1553. Cigarette smoke extract inhibits fluid and HCO3 activity in guinea pig pancreatic ductal cells
secretion and CFTR
ria Venglovecz 2, Krisztina To th 1, Andrea Schnúr 1, Petra Pallagi 1, Vikto n Rakonczay, Jr. 4, Kata zsef Mal } Csupor 3, Zolta th 1, Jo eth 1, Dezso Emese To 5 5 6 } Cseko , Zsuzsanna Helyes , Peter Hegyi
Abstracts / Pancreatology 16 (2016) S1eS130 1
University of Szeged, First Department of Medicine, Hungary University of Szeged, Department of Pharmacology and Pharmacotherapy, Hungary 3 University of Szeged, Department of Pharmacognosy, Hungary 4 University of Szeged, First Department of Medicine, Department of Pathophysiology, Hungary 5 cs, Department of Pharmacology and University of Pe Pharmacotherapy, Hungary 6 cs, Institute for Translational Medicine & Department University of Pe of Translational Medicine/1st Department of Medicine, MTA-SZTE Lendület Translational Gastroenterology Research Group, University of Szeged, Hungary 2
Introduction: Smoking represents an independent risk factor for the development of chronic pancreatitis (CP). It is well documented that secretion of pancreatic ductal alkaline fluid (which is regulated mostly by anion exchangers and CFTR) is diminished in CP. Aims: In this study, we would like to understand whether smoking has any effects on pancreatic ductal fluid and HCO-3 secretion. Materials & methods: Guinea pigs were exposed to cigarette smoke four times a day for 30 min for 6 weeks. The expression of CFTR was analysed by immunohistochemistry. Intra/interlobular pancreatic ducts were isolated from guinea pig pancreas. Cigarette smoke extract (CSE) was prepared by smoking of 15 cigarettes into 10 ml distilled water by a smoking machine. Three different concentraion (20, 40 and 80mg/ml) were diluted using the stock solution. Intracellular pH was evaluated by microfluorometry. Basal and forskolin-stimulated fluid secretion was measured by video microscopy. CFTR currents were detected by whole cell configuration of patch clamp technique. Results: Cigarette smoking significantly diminished the expression of CFTR and the fluid and HCO-3 secretion in guinea pig pancreas. 40mg/ml CSE decreased HCO-3 secretion via inhibition of Cl-/HCO-3 exchanger activity. CSE dose-dependently decreased forskolin-stimulated fluid secretion in guinea pig pancreatic ducts and forskolin-stimulated Cl- current of CFTR Clchannel (20mg/ml by 44.5 %, 40mg/ml by 69.3 % and 80mg/ml by 81.3%). Conclusion: Smoking represents an independent risk factor for the development of chronic pancreatitis (CP). It is well documented that secretion of pancreatic ductal alkaline fluid (which is regulated mostly by anion exchangers and CFTR) is diminished in CP.
Abstract ID: 1554. Early life exposure to stressors promotes pancreatic carcinogenesis in KC mice Marl ene Dufresne 1, Sarah Gandarillas 1, Hubert Lulka 1, Pierre Dubus 2, Muriel Darnaud ery 3, Anne Gabory 4, Claudine Junien 4, Pierre Cordelier 1, ^me Torrisani 1 ero Michelle Kelly-Irving 5, Cyrille Delpierre 5, J 1
Inserm U1037, Cancer Research Center of Toulouse, France Bordeaux 2, EA2406, France Universite 3 gre e, France INRA UMR1286, Nutrition et Neurologie Inte 4 veloppement et Reproduction, UMR-INRA-ENVA-1198, Biologie du De France 5 Inserm, U1027, France 2
Introduction: Stressful experiences across the lifespan contribute to a variety of disorders including anxiety but also diseases like metabolic syndrome and cancer. Importantly, epidemiological studies suggested that cancer risk can be influenced by exposure to stressful conditions and events early on in life. Aims: To study the kinetic of PDAC development in KC mice that experienced one or two early life stressors by combining or not preconceptional nutritional stress and maternal separation. Materials & methods: One group of Pdx1-Cre females were fed with a control diet (CD) for 4 months and a second group received a high fat diet (HFD) for 2 months to induce obesity and then the CD for 2 months to induce weight loss. Females were then bred to LSL-KrasG12D+/- males to generate F1 KC. In maternal separation stressed groups, mouse pups were separated from their mother 3 hours per day from day 2 after birth and to
S23
day 14. Four groups of F1 KC were formed: from HFD mother ± separation, from CD mother ± separation. The pancreas phenotype was analyzed at 8 months of age. Results: Pre-conceptional HFD and/or maternal separation increased the density of preneoplastic lesions in the pancreas of F1 KC mice. We quantified more high-grade PanIN lesions and larger acinar-to-ductal metaplasia in the pancreas of stressed KC mice. Pre-conceptional HFD increased the size of low-grade PanIN and this of high-grade PanIN only when combined to maternal separation. Conclusion: Preconception maternal obesity and/or early life maternal separation promotes pancreatic carcinogenesis in KC mice.
Abstract ID: 1557. Serotonin-selective reuptake inhibitors (SSRI) as a novel approach to counteract pancreatic ductal adenocarcinoma progression Enrica Saponara 1, Gitta Seleznik 1, Raphael Buzzi 1, Irene Esposito 2, Johanna Bushmann 3, Francesco Baschieri 4, Hesso Farhan 5, Michele Visentin 6, Rolf Graf 1, Sabrina Sonda 1 1
Swiss HPB Center, Visceral & Transplantation Surgery, University Hospital Zurich, Switzerland 2 Institut für Pathologie, Universit€ atsklinikum Düsseldorf, Germany 3 Division of Reconstructive Surgery, Switzerland 4 rologie Gustave Roussy, Villejuif, France Institut de Cance 5 Department of Biology, University of Konstanz, Biotechnology Institute Thurgau, Switzerland 6 Department of Clinical Pharmacology and Toxicology, University Hospital Zurich, Switzerland Introduction: Pancreatic ductal adenocarcinoma (PDAC) is classified as one of the most recalcitrant amongst human malignancies due to its remarkable drug resistance. Nevertheless, little is known regarding the critical processes orchestrating PDAC progression, i.e. pancreatic intraepithelial neoplasia, accumulation of a dense stroma, angiogenesis and inflammatory environment. Aims: We previously demonstrated the role of serotonin (5-HT) in pancreatic inflammation and fibrotic processes during pancreatitis. We now aim to investigate the role of 5-HT in cancer progression and to propose potential new pharmacological strategies to counteract PDAC progression. Materials & methods: Transgenic mice harboring KrasG12D mutation, highly invasive human pancreatic cancer cell line (Panc1), human fibroblasts (HFF) and chick chorioallantoic membrane assay (CAM) were utilized to test the therapeutic potential of serotonin-selective reuptake inhibitors (SSRI). Transcriptome sequencing and imaging assays were used to characterize the effect of SSRI on Panc1 and HFF cell line. Results: Prolonged pharmacological treatment of KrasG12D mice with SSRI significantly reduced pre-malignant acinar cell transformation, along with a blunted desmoplastic stroma and inflammatory cell infiltration. Transcriptome analysis of Panc1 cells revealed that SSRI strongly interfered with cell cycle progression and microtubule assembly, concomitant with increased autophagy. Similar features were observed in SSRI-treated HFF cells displaying concomitant impaired cell motility. Finally, SSRI significantly reduced vessel number and diameter of highly vascularized CAM surfaces. Conclusion: Our data suggest that FDA-approved SSRI display alternative and unexpected clinical implications targeting both pre-malignant lesions and tumor micro-environment. Therefore, SSRI, in combination with chemotherapeutics, may constitute a novel approach to counteract PDAC progression.
Abstract ID: 1559. Characterization of pancreatic ductal fluid and bicarbonate secretion in wild type ferrets n th 1, Petra Pallagi 1, Jo zsef Mal ria Venglovecz 2, Zolta Emese To eth 1, Vikto 3 4 Rakonczay, Jr. , Peter Hegyi