Acinar specific TGF-β signaling regulates acinar cell regeneration
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EMT and stemness were assessed in human and murine pancreatic cancer models using migration/invasion and spheroid assay. Results: Here, we identified antidromic NFATc1 and p53 transcrip- tional network control over EMT and stemness. We show that p53 acti- vation prevents cells from EMT in a miR200c dependent manner. However, disruption of the tumor suppressor pathway enables NFATc1/Sox2 chro- matin complex formation and transcription of EMT programmes, resulting in highly invasive and metastatic PDACs. Finally, re-expression of miR200c or NFATc1 inactivation suppresses EMT/stemness genes and re-sensitizes PDAC to chemotherapy. Conclusion: Antidromic NFATc1 and p53 signaling pathways control key features of cellular plasticity and tumor progression at the level of gene transcription. These findings implicate key roles for NFATc1 in transcrip- tional regulation of differentiation and self-renewal in PDAC. S-5 Abstract id: 247. Serotonin regulates progenitor cell-based but not clonal regeneration in the adult pancreatic acinar cell Enrica Saponara, Sabrina Sonda, Kamile Grabliauskaite, Theresia Reding, Rolf Graf. Swiss HPB Center, Visceral & Transplantation Surgery, University Hospital Zurich, Switzerland Introduction: Progenitor cell-based regeneration of acinar cells is activated during cerulein-induced pancreatitis. This process requires acinar de-differentiation via secretion of zymogen content, followed by expression of progenitor cell markers and formation of acinar-to-ductal metaplasia (ADM). Clonal regeneration without loss of zymogen content and cell de-differentiation is observed following 60% pancreatectomy. Aims: We previously showed that serotonin (5-HT) is essential for acinar cell secretion with a strong impact on the pathophysiology of acute pancreatitis; we now investigate the role of 5-HT in pancreatic regeneration. Patients & methods: Cerulein-induced pancreatitis and 60% pancre- atectomy were performed in wild type (WT) and tryptophan hydroxylase- 1 knock-out (TPH1-/-) mice, with reduced peripheral levels of 5-HT. The regenerative potential of pancreatic acinar cells was evaluated in vivo, over a period of two weeks. Results: After experimental pancreatitis, WT mice showed an early up- regulation of 5-HT2A/5-HT2B receptors, amylase secretion and progenitor cell marker expression, indicative of acinar de-differentiation. These events were followed by the appearance of ADM lesions and acinar cell proliferation. Conversely, in TPH1-/- mice these early parameters were blunted and consequent formation of ADM and proliferation of acinar cells were inhibited. After 60% pancreatectomy, clonal regeneration of differ- entiated acinar cell was comparable in the two strains. Similarly to what was observed in vivo, 5-HT2A/5-HT2B receptor agonist promoted prolif- eration only in de-differentiated but not in dexamethasone-differentiated AR42J cells. Conclusion: Our results indicate that 5-HT, likely via 5-HT2A/5-HT2B receptors, modulates progenitor cell-based but not clonal regeneration. Current investigations aim to elucidate the role of 5-HT in pancreatic proliferation induced by primary mitogen administration. S-6 Abstract id: 19. Effect of cystathionine-gamma-lyase gene deletion in caerulein- induced acute pancreatitis in the mouse Madhav Bhatia, Abel Ang, Jack Rivers, Akhil Hegde. University of Otago, Christchurch, New Zealand Introduction: Hydrogen sulfide (H 2 S) has been reported to be involved in the signaling of the inflammatory response; however there are differing views as to whether it is pro- or anti-inflammatory. Aims: The present study was aimed to investigate if endogenously synthesized H 2 S via cystathionine-g-lyase (CSE) plays a pro- or anti-in- flammatory role in caerulein-induced acute pancreatitis Materials & methods: Acute Pancreatitis was induced in CSE knockout (CSE -/- ) and wildtype (CSE þ/þ ) mice by hourly caerulein injections (50 mg/ kg) for 10 hours. Mice were sacrificed 1 h after the last caerulein injection. Blood, pancreas and lung tissues were collected and processed to measure the plasma amylase, plasma H 2 S, myeloperoxidase (MPO) activities and prostaglandin E 2 and cytokine levels in pancreas and lung. Results: CSE -/- mice showed significantly less local pancreatic damage as well as acute pancreatitis-associated lung injury in comparison to the CSE þ/þ mice. There were also lower levels of pancreatic eicosanoid and chemokines in the CSE -/- mice when compared with CSE þ/þ mice. Addi- tionally, in CSE þ/þ mice, there was a greater level of pancreatic CSE expression and sulfide synthesizing activity in caerulein-induced when compared to the saline control. When comparing the two saline treated control groups, the CSE -/- mice showed significantly less pancreatic H 2 S synthesizing activity relative to the CSE þ/þ mice. Conclusion: These data provide evidence that endogenous H 2 S generated by CSE plays a key pro-inflammatory role in caerulein-induced pancreatitis and its genetic deletion affords significant protection against acute pancreatitis and associated lung injury. S-7 Abstract id: 214. Acinar specific TGF-b signaling regulates acinar cell regenerationCate- gory: Basic science - chronic pancreatitis. Kamile Grabliauskaite, Theresia Reding, Enrica Saponara, Sabrina Sonda, Rolf Graf. Hospital Zurich, Switzerland Introduction: TGF-b signaling is implicated in many pathophysiolog- ical functions of pancreatic cells, including regulation of regeneration and fibrosis. However, the function of TGF-b signaling is strongly context- dependent and an acinar cell specific role of this molecule in modulating regeneration has not been completely investigated before. Aims: In this study we aim to determine the contribution of TGFb signaling to acinar cell proliferation during pancreatitis by using mice deficient in TGFb receptor II (TGFbRII fl/fl ) exclusively in acinar cells. Materials & methods: Pancreatitis was induced in control and PTF1b- Cre Tg ; TGFbRII fl/fl mice by multiple injections of cerulein. The expression of proliferation markers, cell cycle regulators, and the severity of tissue inflam- mation and fibrosis were analysed by immunohistochemistry and qRT-PCR. Results: Ki67 and BrdU analyses revealed increased number of prolif- erating acinar cells in TGFbRII fl/fl mice. Surprisingly, immunohistochemistry showed that expression of the cell cycle inhibitor p21WAF1/Cip1, one of the main targets of TGFb signaling, was comparable in WT and TGFbRII fl/fl mice. However, the expression of the cell cycle inhibitor p16INK4a increased only in control animals. In addition, we observed extended formation of acinar- to-ductal metaplasia, higher stellate cells activation and stronger fibrosis in TGFbRII fl/fl mice. Also, analysis of pan-leukocyte infiltration demonstrated stronger inflammation in TGFbRII fl/fl mice compared to control animals. Conclusion: Our data revealed that TGFb signaling inhibits activation of acinar cell cycle and prevents excessive ADM formation. Additionally, the loss of TGFb signaling in acinar cells potentiates fibrogenic processes during pancreatitis, suggesting the existence of a regulatory feedback between acinar and stellate cells. S-8 Abstract id: 147. Pancreatic stellate cells promote the hapto-migration of pancreatic cancer cells through collagen I mediated activation of alpha2beta1 in- tegrin pathway Jing Lu 1 , Shaoxia Zhou 1 , Marco Siech 2 , Hansjoerg Habisch 1 , Thomas Seufferlein 3 , Max. Bachem 1 . Abstracts / Pancreatology 13 (2013) S2–S98 S91
Transcript of Acinar specific TGF-β signaling regulates acinar cell regeneration
Abstracts / Pancreatology 13 (2013) S2–S98 S91
EMT and stemness were assessed in human and murine pancreatic cancermodels using migration/invasion and spheroid assay.
Results: Here, we identified antidromic NFATc1 and p53 transcrip-tional network control over EMT and stemness. We show that p53 acti-vation prevents cells from EMT in amiR200c dependent manner. However,disruption of the tumor suppressor pathway enables NFATc1/Sox2 chro-matin complex formation and transcription of EMT programmes, resultingin highly invasive and metastatic PDACs. Finally, re-expression of miR200cor NFATc1 inactivation suppresses EMT/stemness genes and re-sensitizesPDAC to chemotherapy.
Conclusion: Antidromic NFATc1 and p53 signaling pathways controlkey features of cellular plasticity and tumor progression at the level of genetranscription. These findings implicate key roles for NFATc1 in transcrip-tional regulation of differentiation and self-renewal in PDAC.
S-5 Abstract id: 247.
Serotonin regulates progenitor cell-based but not clonal regenerationin the adult pancreatic acinar cell
Enrica Saponara, Sabrina Sonda, Kamile Grabliauskaite, TheresiaReding, Rolf Graf.
Swiss HPB Center, Visceral & Transplantation Surgery, UniversityHospital Zurich, Switzerland
Introduction: Progenitor cell-based regeneration of acinar cells isactivated during cerulein-induced pancreatitis. This process requiresacinar de-differentiation via secretion of zymogen content, followed byexpression of progenitor cell markers and formation of acinar-to-ductalmetaplasia (ADM). Clonal regeneration without loss of zymogen contentand cell de-differentiation is observed following 60% pancreatectomy.
Aims:Wepreviously showed that serotonin (5-HT) is essential for acinarcell secretion with a strong impact on the pathophysiology of acutepancreatitis; we now investigate the role of 5-HT in pancreatic regeneration.
Patients & methods: Cerulein-induced pancreatitis and 60% pancre-atectomy were performed in wild type (WT) and tryptophan hydroxylase-1 knock-out (TPH1-/-) mice, with reduced peripheral levels of 5-HT. Theregenerative potential of pancreatic acinar cells was evaluated in vivo, overa period of two weeks.
Results: After experimental pancreatitis, WTmice showed an early up-regulation of 5-HT2A/5-HT2B receptors, amylase secretion and progenitorcell marker expression, indicative of acinar de-differentiation. Theseevents were followed by the appearance of ADM lesions and acinar cellproliferation. Conversely, in TPH1-/- mice these early parameters wereblunted and consequent formation of ADM and proliferation of acinar cellswere inhibited. After 60% pancreatectomy, clonal regeneration of differ-entiated acinar cell was comparable in the two strains. Similarly to whatwas observed in vivo, 5-HT2A/5-HT2B receptor agonist promoted prolif-eration only in de-differentiated but not in dexamethasone-differentiatedAR42J cells.
Conclusion: Our results indicate that 5-HT, likely via 5-HT2A/5-HT2Breceptors, modulates progenitor cell-based but not clonal regeneration.Current investigations aim to elucidate the role of 5-HT in pancreaticproliferation induced by primary mitogen administration.
S-6 Abstract id: 19.
Effect of cystathionine-gamma-lyase gene deletion in caerulein-induced acute pancreatitis in the mouse
Madhav Bhatia, Abel Ang, Jack Rivers, Akhil Hegde.
University of Otago, Christchurch, New Zealand
Introduction: Hydrogen sulfide (H2S) has been reported to be involvedin the signaling of the inflammatory response; however there are differingviews as to whether it is pro- or anti-inflammatory.
Aims: The present study was aimed to investigate if endogenouslysynthesized H2S via cystathionine-g-lyase (CSE) plays a pro- or anti-in-flammatory role in caerulein-induced acute pancreatitis
Materials &methods: Acute Pancreatitis was induced in CSE knockout(CSE-/-) and wildtype (CSEþ/þ) mice by hourly caerulein injections (50 mg/kg) for 10 hours. Mice were sacrificed 1 h after the last caerulein injection.Blood, pancreas and lung tissues were collected and processed to measurethe plasma amylase, plasma H2S, myeloperoxidase (MPO) activities andprostaglandin E2 and cytokine levels in pancreas and lung.
Results: CSE-/- mice showed significantly less local pancreatic damageas well as acute pancreatitis-associated lung injury in comparison to theCSEþ/þ mice. There were also lower levels of pancreatic eicosanoid andchemokines in the CSE-/-mice when compared with CSEþ/þ mice. Addi-tionally, in CSEþ/þ mice, there was a greater level of pancreatic CSEexpression and sulfide synthesizing activity in caerulein-induced whencompared to the saline control. When comparing the two saline treatedcontrol groups, the CSE-/- mice showed significantly less pancreatic H2Ssynthesizing activity relative to the CSEþ/þ mice.
Conclusion: These data provide evidence that endogenous H2Sgenerated by CSE plays a key pro-inflammatory role in caerulein-inducedpancreatitis and its genetic deletion affords significant protection againstacute pancreatitis and associated lung injury.
S-7 Abstract id: 214.
Acinar specific TGF-b signaling regulates acinar cell regenerationCate-gory: Basic science - chronic pancreatitis.
Kamile Grabliauskaite, Theresia Reding, Enrica Saponara, SabrinaSonda, Rolf Graf.
Hospital Zurich, Switzerland
Introduction: TGF-b signaling is implicated in many pathophysiolog-ical functions of pancreatic cells, including regulation of regeneration andfibrosis. However, the function of TGF-b signaling is strongly context-dependent and an acinar cell specific role of this molecule in modulatingregeneration has not been completely investigated before.
Aims: In this study we aim to determine the contribution of TGFbsignaling to acinar cell proliferation during pancreatitis by using micedeficient in TGFb receptor II (TGFbRII fl/fl) exclusively in acinar cells.
Materials & methods: Pancreatitis was induced in control and PTF1b-CreTg; TGFbRIIfl/fl mice by multiple injections of cerulein. The expression ofproliferation markers, cell cycle regulators, and the severity of tissue inflam-mation and fibrosis were analysed by immunohistochemistry and qRT-PCR.
Results: Ki67 and BrdU analyses revealed increased number of prolif-erating acinar cells inTGFbRIIfl/flmice. Surprisingly, immunohistochemistryshowed that expression of the cell cycle inhibitor p21WAF1/Cip1, one of themain targets of TGFb signaling, was comparable inWTand TGFbRIIfl/flmice.However, the expression of the cell cycle inhibitor p16INK4a increased onlyin control animals. In addition, we observed extended formation of acinar-to-ductal metaplasia, higher stellate cells activation and stronger fibrosis inTGFbRIIfl/fl mice. Also, analysis of pan-leukocyte infiltration demonstratedstronger inflammation in TGFbRIIfl/fl mice compared to control animals.
Conclusion: Our data revealed that TGFb signaling inhibits activationof acinar cell cycle and prevents excessive ADM formation. Additionally,the loss of TGFb signaling in acinar cells potentiates fibrogenic processesduring pancreatitis, suggesting the existence of a regulatory feedbackbetween acinar and stellate cells.
S-8 Abstract id: 147.
Pancreatic stellate cells promote the hapto-migration of pancreaticcancer cells through collagen I mediated activation of alpha2beta1 in-tegrin pathway
Jing Lu 1, Shaoxia Zhou 1, Marco Siech 2, Hansjoerg Habisch 1, ThomasSeufferlein 3, Max. Bachem 1.
