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MODS, APACHE scores versus novel model of acute pancreatitis assessment in prediction of disease outcome
Abstracts / Pancreatology 15 (2015) S1eS141
975. Inhibition of MAPKs and type IV phosphodiesterase during the early phase of acute pancreatitis: A shotgun proteomics approach ndez 1, Domitille Schvartz 2, Violeta García-Herna n Calvo 3, Jean-Charles Sanchez 2, nchez-Bernal 1, Jos e Julia Carmen Sa 1 Jesús Sanchez-Yagüe 1 Department of Biochemistry and Molecular Biology, University of n Biome dica de Salamanca), Spain Salamanca, IBSAL (Inst. Investigacio 2 Translational Biomarker Group, Department of Human Protein Sciences, University Medical Center, Geneva, Switzerland 3 Department of Physiology and Pharmacology, University of n Biome dica de Salamanca), Spain Salamanca, IBSAL (Inst. Investigacio
Introduction: Early changes in protein expression and signaling mechanisms (MAPKs cascade or the cAMP pathway) in acute pancreatitis (AP) play important roles in the further development of the disease. Therefore, proteomic analyses of pancreatic subcellular fractions might be very useful for deciphering AP at the molecular level. Aims: We reported here a Tandem Mass Tag (TMT) proteomics analysis of the soluble fraction of the pancreas during the early phase of experimental AP and under MAPKs or type IV phosphodiesterase (PDEIV) inhibition. Materials & methods: The early phase of AP was induced in rats by two subcutaneous injections of 20 mg of cerulein (Cer)/kg body weight at hourly intervals. PDEIV and the MAPKs cascade were inhibited by rolipram and SP600125 or PD98059, respectively, before the induction of AP. TMT6 analysis were designed to characterize each experimental condition in a cross-sectional study. Changes in protein expression were validated by immunoblotting. Results: Out of the around 540 identified proteins, 298 were differentially expressed during the early phase of AP (Isobar, p < 0.05). Also, we identified 27 (SP600125), 39 (PD98059) or 38 (rolipram) proteins differentially regulated by the distinct pretreatments compared to the Cer class. These proteins are mainly involved in protein and membrane trafficking, apoptosis, mitochondrial function, calcium signaling and cytoskeleton impairment. We are currently characterizing novel candidates that might be relevant to AP pathogenesis. Conclusion: These data should provide valuable information for unraveling the early pathophysiologic mechanisms of Cer-induced AP and might unmask new potential biomarkers of the disease. Support: FEDER-FIS (PI09/1075). FPU-MECD(AP2010-2350). EMBO Short-Term fellowship (ASTF523 e 2014).
1237. Kynurenic acid and its novel analogue SZR-72 attenuate the severity of experimental acute necrotizing pancreatitis zs Kui 1, Eszter S. Korma nyos 1, B nyi 2, Zsolt Balla 1, Bala ela Iva 3 4 1 € Laszlo Vecsei , Ferenc Fülop , Tibor Wittmann , Peter Hegyi 1, 5, n Rakonczay, Jr. 1 Zolta 1
First Department of Medicine, University of Szeged, Hungary Department of Pathology, University of Szeged, Hungary 3 Department of Neurology, University of Szeged, Hungary 4 Institute of Pharmaceutical Chemistry, University of Szeged, Hungary 5 MTA-SZTE Translational Gastroenterology Research Group, Szeged, Hungary 2
Introduction: The pathogenesis of acute pancreatitis (AP) is not well understood and the disease has no specific therapy. There is evidence that the blood brain barrier (BBB) permeable L-kynurenic acid (KYN) analogue SZR-72 has immune modulatory roles in several inflammatory diseases. Aims: We investigated the effects of KYN and SZR-72 on experimental AP. Patients & methods: In the AP groups, male SPRD rats were injected intraperitoneally (i.p.) with 3 g/kg L-ornithine 1 hour after the administration of physiological saline(PS) (n ¼ 6e8) or 30e300 mg/kg SZR-72 (n ¼ 6e8) or KYN (n ¼ 6e8). Control animals were injected i.p. with PS instead of L-ornithine or 30e300 mg/kg SZR-72 or KYN (n ¼ 6e8). Animals
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were sacrificed at 24 hours. Laboratory [serum amylase-, pancreatic myeloperoxidase activity, pancreatic dry/wet weight ratio] and histological parameters were measured to evaluate disease severity. Results: The injection of rats with L-ornithine significantly increased the measured laboratory and histological parameters vs. the control groups. The administration of 30e300 mg/kg SZR-72 or KYN did not influence the examined parameters in the control groups. Pre-treatment of AP rats with 30 mg/kg SZR-72 or KYN did not have affect on disease severity. However, all measured laboratory and histological parameters were significantly reduced in AP animals in response to treatment with 300 mg/kg SZR-72 or KYN. Conclusion: Our experiments demonstrated that SZR-72 and KYN have a dose-dependent protective effect on L-ornithine-induced AP. Since KYN is non BBB permeable and both KYN and SZR-72 exerted a beneficial effect on AP severity, this beneficial effect seems to peripheral. Further investigations are needed to determine the mechanism of SZR-72 action.
1240. The role of endogenous and exogenous corticosteroids in the pathogenesis of acute experimental pancreatitis zs Kui 1, Zsolt Balla 1, Eszter S. Korma nyos 1, B nyi 2, Bala ela Iva n Rakonczay 1 P eter Hegyi 1, 3, Zolta 1
First Department of Medicine University of Szeged, Hungary Department of Pathology University of Szeged, Hungary 3 MTA-SZTE Translational Gastroenterology Research Group, Szeged, Hungary 2
Introduction: Acute pancreatitis (AP) is one of the most common emergency gastroenterological diseases. The mortality of AP with organ failure is unacceptably high, 25e40%. The pathomechanism of the disease is not well understood, it has no specific therapy. Administration of corticosteroids is widespread in the treatment of many inflammatory diseases, but the use of corticosteroids in AP is debated. Aims: We aimed to investigate the role of endogenous and exogenous corticosteroids in experimental AP. Materials & methods: AP was induced in SPRD rats by intraperitoneal (i.p.) injection of 3 g/kg L-ornithine (n ¼ 8), control animals (n ¼ 6) received physiological saline (PS) instead of the basic amino acid. Animals were sacrificed 24 hours after AP induction. Endogenous corticosteroid levels were increased (as measured by ELISA) by immobilization stress; exogenous corticosteroid (2 mg/kg betamethasone) was administered intramuscularly before the injection of L-ornithine or PS (n ¼ 6e8). Laboratory and histological parameters were measured to evaluate the severity of AP. Results: L-ornithine injection induced necrotizing AP. Elevated serum corticosteroid concentrations were confirmed in stressed vs non-stressed group. Histological scores (oedema, leukocyte infiltration, cell damage) and laboratory parameters (serum amylase-, pancreatic myeloperoxidase activity) were significantly lower in stressed vs. non-stressed animals with AP. We also found milder AP severity as measured by laboratory and histological parameters after the exogenous administration of corticosteroid vs. vehicle-treated AP group. Conclusion: Corticosteroids ameliorated the severity of AP in rats and thus may have a beneficial effect in experimental AP, but further investigation is needed to determine the exact mechanism.
1243. MODS, APACHE scores versus novel model of acute pancreatitis assessment in prediction of disease outcome Shamil Galeev 1, Yakubbay Abdullaev 2, Rubtsov Michail 1 1 2
Saint Luke's Clinical Hospital, Russia Institute of Surgery n.a. Vachidov, Uzbekistan
Introduction: There are few relevant methods for predicting fatal outcome and prolonged ICU stay in acute pancreatitis. According to
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Abstracts / Pancreatology 15 (2015) S1eS141
literature data, most scoring systems serve as predictors of severity and outcome simultaneously. Aims: To compare predictive strength (in point of outcome prediction) of MODS and APACHE scores with novel model of pancreatitis assessment. Materials & methods: In retrospective study which included 89 patients with acute pancreatitis during 9 year period, outcome prediction model was developed. It included 5 variables, conjoined into 3 groups: 1. early pancreatitis-specific organ failure (PO2/FiO2 < 250, creatinine >150 mcmol/l, pressure-adjusted heart rate 10); 2. CT data (extended pancreatic necrosis); 3. infected necrosis. Outcome prediction accuracy of this model amounted to 88,3% (sensitivity-93,3%; specificity-84,2%). Detection of 2 or more factors according to abovementioned score was closely associated with persistence of organ failure, and death. Retrospective analysis of 31 patients allowed us to compare diagnostic accuracy of conventional (APACHE, MODS score) and our novel system. Results: Novel prognostic model proved itself as more accurate then MODS and APACHE in prediction of negative outcome (p < 0,05). Conclusion: Novel prognostic model has improved outcome predictive value compared with conventional scoring systems. Further studies are necessary to optimize early outcome prediction in severe acute pancreatitis.
1250. Inactivation of TGF-b receptor II signalling in pancreatic epithelial cells promotes acinar cell replication and acinar-to-ductal metaplasia formation during pancreatitis Kamile Grabliauskaite, Enrica Saponara, Theresia Reding, Sabrina Sonda, Rolf Graf Department of Visceral and Transplantation Surgery, University Hospital Zurich, Switzerland Introduction: TGF-b signalling is implicated in pancreatic regeneration and fibrosis. However, the function of TGF-b signalling is strongly contextdependent and an acinar cell specific role of this molecule in modulating regeneration has not been completely investigated before. Aims: In this study we aimed to determine the contribution of TGF-b signalling to acinar cell regeneration during pancreatitis by using mice deficient in TGF-b receptor II (TGFbRIIfl/fl) in epithelial cells. Materials & methods: Pancreatitis was induced in control and PTF1aCreTg; TGFbRIIfl/fl mice by multiple injections of cerulein. The expression of proliferation markers, cell cycle regulators, and the severity of tissue inflammation and fibrosis were analysed by immunohistochemistry, western blotting and qRT-PCR. Results: At the onset of pancreatitis, serum levels of amylase and lipase and pancreatic tissue damage were comparable in control and TGFbRIIfl/fl mice. However, after prolonged exposure to cerulein, lack of TGF-b receptor II resulted in an increased number of proliferating acinar cells and reduced expression of the cell cycle inhibitor p16INK4a. In addition, TGFbRIIfl/fl mice showed extended acinar-to-ductal metaplasia together with increased activation of EGFR signalling. Finally, after one week of cerulein treatment, TGFbRIIfl/fl mice presented higher stellate cells activation and stronger fibrosis accompanied by robust inflammation. Conclusion: Our data revealed that activation of TGF-b signalling in epithelial cells during pancreatitis does not mediate the initial acinar cell damage, however it prevents excessive activation of acinar cell cycle and limits ADM formation. Additionally, the potentiation of fibrosis following loss of TGF-b signalling in acinar cells, suggests the existence of a regulatory feedback between epithelial and stromal cells.
1054. Pancreatic stellate cells are activated by cigarette smoke components and alcohol Minoti Apte, Zhihong Xu, Alexandra Lee, Srinivasa Pothula, Romano Pirola, Jeremy Wilson Pancreatic Research Group, SWS Clinical School, University of New South Wales, and Ingham Institute for Applied Medical Research, Australia Introduction: Smoking has been reported to accelerate progression of alcohol chronic pancreatitis (CP), as evidenced by early calcification and fibrosis. Activated pancreatic stellate cells (PSCs) are the key cells responsible for production of pancreatic fibrosis, but the effects of smoking ± alcohol on PSCs are not fully elucidated. Aims: To examine the effects of smoke components±alcohol on expression of i) a7 nicotinic acetylcholine receptor (a7nAChR, the major receptor for cigarette smoke components nicotine and nicotine-derived nitrosamine ketone, NNK), ii) collagen I, iii) Light Chain 3B (LC3B, a marker of autophagy) in human PSCs (hPSCs) in vitro. Materials & methods: Human PSCs (n¼4-5 separate cell preparations) were exposed for 48 hours to clinically relevant concentrations of ethanol 10mM (E10) ± cigarette smoke extract (CSE) 40ng/mL or nicotine 1mM or NNK 100mM. a7nAChR and LC3B expression assessed by western blotting; collagen I measured by ELISA. Results: In hPSCs, i) The expression of a7nAChR was significantly induced by CSEþE10 (199.6±60.4% of vehicle control, n¼4, p<0.05), ii) The expression of collagen I was significantly induced by NNKþE10 (0.1389±0.044 vs vehicle control 0.0319±0.014, n¼4, p<0.05), iii) Ratio of LC3BII to LC3BI was significantly increased by CSEþE10 and NNKþE10, thus suggesting increased autophagosome formation in hPSCs (148.4±11.87% and 151.7±10.73% of vehicle control respectively, n¼5, p<0.05). Conclusion: Smoking components when combined with alcohol increased expression of a7nAChR and collagen I, and induced autophagosome formation in hPSCs. These effects may provide a possible mechanism for smoking induced progression of alcoholic pancreatic fibrosis.
1058. NFATc1-EGFR loop controls Sox9 promoter for acinar ductal metaplasia €nig, Nai-Ming Chen, Garima Singh, Elisabeth Hessmann, Alex Ko Volker Ellenrieder Gastroenterology, Germany Introduction: Mutations of KRAS are important drivers of pancreatic cancer progression. One of the main risk factors for the development of PDA in humans is chronic pancreatitis that induces the expression of EGFR and nuclear factor of activated T cells. Recent evidence has shown KRASdriven and pancreatitis-associated pancreatic tumor initiation is dependent on EGFR. The signaling pathways downstream of EGFR that mediate ADM are less clear. Aims: To explore the interaction of NFATc1 and EGFR signalling in acinar ductal metaplasia in the presence of Kras mutation.
975. Inhibition of MAPKs and type IV phosphodiesterase during the early phase of acute pancreatitis: A shotgun proteomics approach ndez 1, Domitille Schvartz 2, Violeta García-Herna n Calvo 3, Jean-Charles Sanchez 2, nchez-Bernal 1, Jos e Julia Carmen Sa 1 Jesús Sanchez-Yagüe 1 Department of Biochemistry and Molecular Biology, University of n Biome dica de Salamanca), Spain Salamanca, IBSAL (Inst. Investigacio 2 Translational Biomarker Group, Department of Human Protein Sciences, University Medical Center, Geneva, Switzerland 3 Department of Physiology and Pharmacology, University of n Biome dica de Salamanca), Spain Salamanca, IBSAL (Inst. Investigacio
Introduction: Early changes in protein expression and signaling mechanisms (MAPKs cascade or the cAMP pathway) in acute pancreatitis (AP) play important roles in the further development of the disease. Therefore, proteomic analyses of pancreatic subcellular fractions might be very useful for deciphering AP at the molecular level. Aims: We reported here a Tandem Mass Tag (TMT) proteomics analysis of the soluble fraction of the pancreas during the early phase of experimental AP and under MAPKs or type IV phosphodiesterase (PDEIV) inhibition. Materials & methods: The early phase of AP was induced in rats by two subcutaneous injections of 20 mg of cerulein (Cer)/kg body weight at hourly intervals. PDEIV and the MAPKs cascade were inhibited by rolipram and SP600125 or PD98059, respectively, before the induction of AP. TMT6 analysis were designed to characterize each experimental condition in a cross-sectional study. Changes in protein expression were validated by immunoblotting. Results: Out of the around 540 identified proteins, 298 were differentially expressed during the early phase of AP (Isobar, p < 0.05). Also, we identified 27 (SP600125), 39 (PD98059) or 38 (rolipram) proteins differentially regulated by the distinct pretreatments compared to the Cer class. These proteins are mainly involved in protein and membrane trafficking, apoptosis, mitochondrial function, calcium signaling and cytoskeleton impairment. We are currently characterizing novel candidates that might be relevant to AP pathogenesis. Conclusion: These data should provide valuable information for unraveling the early pathophysiologic mechanisms of Cer-induced AP and might unmask new potential biomarkers of the disease. Support: FEDER-FIS (PI09/1075). FPU-MECD(AP2010-2350). EMBO Short-Term fellowship (ASTF523 e 2014).
1237. Kynurenic acid and its novel analogue SZR-72 attenuate the severity of experimental acute necrotizing pancreatitis zs Kui 1, Eszter S. Korma nyos 1, B nyi 2, Zsolt Balla 1, Bala ela Iva 3 4 1 € Laszlo Vecsei , Ferenc Fülop , Tibor Wittmann , Peter Hegyi 1, 5, n Rakonczay, Jr. 1 Zolta 1
First Department of Medicine, University of Szeged, Hungary Department of Pathology, University of Szeged, Hungary 3 Department of Neurology, University of Szeged, Hungary 4 Institute of Pharmaceutical Chemistry, University of Szeged, Hungary 5 MTA-SZTE Translational Gastroenterology Research Group, Szeged, Hungary 2
Introduction: The pathogenesis of acute pancreatitis (AP) is not well understood and the disease has no specific therapy. There is evidence that the blood brain barrier (BBB) permeable L-kynurenic acid (KYN) analogue SZR-72 has immune modulatory roles in several inflammatory diseases. Aims: We investigated the effects of KYN and SZR-72 on experimental AP. Patients & methods: In the AP groups, male SPRD rats were injected intraperitoneally (i.p.) with 3 g/kg L-ornithine 1 hour after the administration of physiological saline(PS) (n ¼ 6e8) or 30e300 mg/kg SZR-72 (n ¼ 6e8) or KYN (n ¼ 6e8). Control animals were injected i.p. with PS instead of L-ornithine or 30e300 mg/kg SZR-72 or KYN (n ¼ 6e8). Animals
S9
were sacrificed at 24 hours. Laboratory [serum amylase-, pancreatic myeloperoxidase activity, pancreatic dry/wet weight ratio] and histological parameters were measured to evaluate disease severity. Results: The injection of rats with L-ornithine significantly increased the measured laboratory and histological parameters vs. the control groups. The administration of 30e300 mg/kg SZR-72 or KYN did not influence the examined parameters in the control groups. Pre-treatment of AP rats with 30 mg/kg SZR-72 or KYN did not have affect on disease severity. However, all measured laboratory and histological parameters were significantly reduced in AP animals in response to treatment with 300 mg/kg SZR-72 or KYN. Conclusion: Our experiments demonstrated that SZR-72 and KYN have a dose-dependent protective effect on L-ornithine-induced AP. Since KYN is non BBB permeable and both KYN and SZR-72 exerted a beneficial effect on AP severity, this beneficial effect seems to peripheral. Further investigations are needed to determine the mechanism of SZR-72 action.
1240. The role of endogenous and exogenous corticosteroids in the pathogenesis of acute experimental pancreatitis zs Kui 1, Zsolt Balla 1, Eszter S. Korma nyos 1, B nyi 2, Bala ela Iva n Rakonczay 1 P eter Hegyi 1, 3, Zolta 1
First Department of Medicine University of Szeged, Hungary Department of Pathology University of Szeged, Hungary 3 MTA-SZTE Translational Gastroenterology Research Group, Szeged, Hungary 2
Introduction: Acute pancreatitis (AP) is one of the most common emergency gastroenterological diseases. The mortality of AP with organ failure is unacceptably high, 25e40%. The pathomechanism of the disease is not well understood, it has no specific therapy. Administration of corticosteroids is widespread in the treatment of many inflammatory diseases, but the use of corticosteroids in AP is debated. Aims: We aimed to investigate the role of endogenous and exogenous corticosteroids in experimental AP. Materials & methods: AP was induced in SPRD rats by intraperitoneal (i.p.) injection of 3 g/kg L-ornithine (n ¼ 8), control animals (n ¼ 6) received physiological saline (PS) instead of the basic amino acid. Animals were sacrificed 24 hours after AP induction. Endogenous corticosteroid levels were increased (as measured by ELISA) by immobilization stress; exogenous corticosteroid (2 mg/kg betamethasone) was administered intramuscularly before the injection of L-ornithine or PS (n ¼ 6e8). Laboratory and histological parameters were measured to evaluate the severity of AP. Results: L-ornithine injection induced necrotizing AP. Elevated serum corticosteroid concentrations were confirmed in stressed vs non-stressed group. Histological scores (oedema, leukocyte infiltration, cell damage) and laboratory parameters (serum amylase-, pancreatic myeloperoxidase activity) were significantly lower in stressed vs. non-stressed animals with AP. We also found milder AP severity as measured by laboratory and histological parameters after the exogenous administration of corticosteroid vs. vehicle-treated AP group. Conclusion: Corticosteroids ameliorated the severity of AP in rats and thus may have a beneficial effect in experimental AP, but further investigation is needed to determine the exact mechanism.
1243. MODS, APACHE scores versus novel model of acute pancreatitis assessment in prediction of disease outcome Shamil Galeev 1, Yakubbay Abdullaev 2, Rubtsov Michail 1 1 2
Saint Luke's Clinical Hospital, Russia Institute of Surgery n.a. Vachidov, Uzbekistan
Introduction: There are few relevant methods for predicting fatal outcome and prolonged ICU stay in acute pancreatitis. According to
S10
Abstracts / Pancreatology 15 (2015) S1eS141
literature data, most scoring systems serve as predictors of severity and outcome simultaneously. Aims: To compare predictive strength (in point of outcome prediction) of MODS and APACHE scores with novel model of pancreatitis assessment. Materials & methods: In retrospective study which included 89 patients with acute pancreatitis during 9 year period, outcome prediction model was developed. It included 5 variables, conjoined into 3 groups: 1. early pancreatitis-specific organ failure (PO2/FiO2 < 250, creatinine >150 mcmol/l, pressure-adjusted heart rate 10); 2. CT data (extended pancreatic necrosis); 3. infected necrosis. Outcome prediction accuracy of this model amounted to 88,3% (sensitivity-93,3%; specificity-84,2%). Detection of 2 or more factors according to abovementioned score was closely associated with persistence of organ failure, and death. Retrospective analysis of 31 patients allowed us to compare diagnostic accuracy of conventional (APACHE, MODS score) and our novel system. Results: Novel prognostic model proved itself as more accurate then MODS and APACHE in prediction of negative outcome (p < 0,05). Conclusion: Novel prognostic model has improved outcome predictive value compared with conventional scoring systems. Further studies are necessary to optimize early outcome prediction in severe acute pancreatitis.
1250. Inactivation of TGF-b receptor II signalling in pancreatic epithelial cells promotes acinar cell replication and acinar-to-ductal metaplasia formation during pancreatitis Kamile Grabliauskaite, Enrica Saponara, Theresia Reding, Sabrina Sonda, Rolf Graf Department of Visceral and Transplantation Surgery, University Hospital Zurich, Switzerland Introduction: TGF-b signalling is implicated in pancreatic regeneration and fibrosis. However, the function of TGF-b signalling is strongly contextdependent and an acinar cell specific role of this molecule in modulating regeneration has not been completely investigated before. Aims: In this study we aimed to determine the contribution of TGF-b signalling to acinar cell regeneration during pancreatitis by using mice deficient in TGF-b receptor II (TGFbRIIfl/fl) in epithelial cells. Materials & methods: Pancreatitis was induced in control and PTF1aCreTg; TGFbRIIfl/fl mice by multiple injections of cerulein. The expression of proliferation markers, cell cycle regulators, and the severity of tissue inflammation and fibrosis were analysed by immunohistochemistry, western blotting and qRT-PCR. Results: At the onset of pancreatitis, serum levels of amylase and lipase and pancreatic tissue damage were comparable in control and TGFbRIIfl/fl mice. However, after prolonged exposure to cerulein, lack of TGF-b receptor II resulted in an increased number of proliferating acinar cells and reduced expression of the cell cycle inhibitor p16INK4a. In addition, TGFbRIIfl/fl mice showed extended acinar-to-ductal metaplasia together with increased activation of EGFR signalling. Finally, after one week of cerulein treatment, TGFbRIIfl/fl mice presented higher stellate cells activation and stronger fibrosis accompanied by robust inflammation. Conclusion: Our data revealed that activation of TGF-b signalling in epithelial cells during pancreatitis does not mediate the initial acinar cell damage, however it prevents excessive activation of acinar cell cycle and limits ADM formation. Additionally, the potentiation of fibrosis following loss of TGF-b signalling in acinar cells, suggests the existence of a regulatory feedback between epithelial and stromal cells.
1054. Pancreatic stellate cells are activated by cigarette smoke components and alcohol Minoti Apte, Zhihong Xu, Alexandra Lee, Srinivasa Pothula, Romano Pirola, Jeremy Wilson Pancreatic Research Group, SWS Clinical School, University of New South Wales, and Ingham Institute for Applied Medical Research, Australia Introduction: Smoking has been reported to accelerate progression of alcohol chronic pancreatitis (CP), as evidenced by early calcification and fibrosis. Activated pancreatic stellate cells (PSCs) are the key cells responsible for production of pancreatic fibrosis, but the effects of smoking ± alcohol on PSCs are not fully elucidated. Aims: To examine the effects of smoke components±alcohol on expression of i) a7 nicotinic acetylcholine receptor (a7nAChR, the major receptor for cigarette smoke components nicotine and nicotine-derived nitrosamine ketone, NNK), ii) collagen I, iii) Light Chain 3B (LC3B, a marker of autophagy) in human PSCs (hPSCs) in vitro. Materials & methods: Human PSCs (n¼4-5 separate cell preparations) were exposed for 48 hours to clinically relevant concentrations of ethanol 10mM (E10) ± cigarette smoke extract (CSE) 40ng/mL or nicotine 1mM or NNK 100mM. a7nAChR and LC3B expression assessed by western blotting; collagen I measured by ELISA. Results: In hPSCs, i) The expression of a7nAChR was significantly induced by CSEþE10 (199.6±60.4% of vehicle control, n¼4, p<0.05), ii) The expression of collagen I was significantly induced by NNKþE10 (0.1389±0.044 vs vehicle control 0.0319±0.014, n¼4, p<0.05), iii) Ratio of LC3BII to LC3BI was significantly increased by CSEþE10 and NNKþE10, thus suggesting increased autophagosome formation in hPSCs (148.4±11.87% and 151.7±10.73% of vehicle control respectively, n¼5, p<0.05). Conclusion: Smoking components when combined with alcohol increased expression of a7nAChR and collagen I, and induced autophagosome formation in hPSCs. These effects may provide a possible mechanism for smoking induced progression of alcoholic pancreatic fibrosis.
1058. NFATc1-EGFR loop controls Sox9 promoter for acinar ductal metaplasia €nig, Nai-Ming Chen, Garima Singh, Elisabeth Hessmann, Alex Ko Volker Ellenrieder Gastroenterology, Germany Introduction: Mutations of KRAS are important drivers of pancreatic cancer progression. One of the main risk factors for the development of PDA in humans is chronic pancreatitis that induces the expression of EGFR and nuclear factor of activated T cells. Recent evidence has shown KRASdriven and pancreatitis-associated pancreatic tumor initiation is dependent on EGFR. The signaling pathways downstream of EGFR that mediate ADM are less clear. Aims: To explore the interaction of NFATc1 and EGFR signalling in acinar ductal metaplasia in the presence of Kras mutation.