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Bridging the gap between bench and bedside: Systematic review of treatment in experimental acute pancreatitis
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Abstracts / Pancreatology 13 (2013) e1–e20
vs. 79.1%). In HC, CP and PC, the combination of three markers (iC3b, ULBP2 and CA 19.9) has better predictive value than CA19.9 alone or combination of ULBP2 and CA19.9 or combination of iC3b and CA19.9 (79.6% vs. 78% vs. 77.8% vs. 79.4%). Conclusion: Our preliminary results show that there is possibility of better predictive value in sensitivity and specificity with a combination of markers rather than a single marker alone. More samples are needed to qualify this finding. Take-home message: A combination of markers may have better predictive value than a single marker alone. Abstract previously presented? no () Any disclosures? no ()
P13. Activated pancreatic stellate cells inhibit CD8+ infiltrate in pancreatic ductal adenocarcinoma via CXCL12 Abasi Ene-Obong, Andrew J. Clear, Alan G. Ramsay, Hemant M. Kocher. Barts Cancer Institute, UK Category: Malignant Background: Pancreatic ductal adenocarcinoma (PDAC) is characterised by a prominent immune cell response. We hypothesized a differential immune cell infiltrate in distinct stromal compartments. Method: Specific tissue microarrays focusing on juxta-tumoural stroma (<100 mm from tumour) from panstroma from various pancreatico-biliary diseases were stained for distinct immune cell markers. Immune cell densities of the stromal compartments were carried out using automated software. Prognostic significance was determined with X-Tile. In vitro and in vivo assays were performed to delineate the mechanistic aspect of the immune infiltrate pattern. Results: PDAC had more immune cell infiltrate when compared to other pancreatico-biliary diseases except for CD8+, which was lower and comparable to that of chronic inflammation (p<0.001). CD3+, CD8+, Foxp3+, CD20+ cells PDAC could not infiltrate the juxta-tumoural stromal compartment whereas myeloperoxidase+, CD4+, CD68+ could do so, in two cohorts (p<0.05). PDAC patients with higher CD8+ densities survived longer than patients with lower densities (p ¼0.03). KPC mice stroma demonstrated similar juxta-tumoral exclusion which could be reversed upon stromal collapse by targeting stellate cells (p<0.05). In vitro migration assays demonstrated increased T-cell migration towards activated pancreatic stellate cells (PSC) compared to quiescent PSC (p¼0.0006), which was abrogated by knockdown of CXCL12 in aPSC (p<0.01). Adhesion assays demonstrated increased adhesion of T-cells to aPSC than qPSC (p¼0.0079). Conclusion: Activated stellate cells hinder the migration of T-cells, specifically, cytotoxic CD8+ T-cells, towards the juxta-tumoural stroma due to secreted CXCL12. Rendering aPSC quiescent reduces CXCL12 production which may enable CD8+ cells to perform tumour cytotoxic functions. Take-home message: Activated pancreatic stellate cells hinder cytotoxic CD8+ T cell infiltrate to the tumour. Rendering activated pancreatic stellate cells quiescent may enable CD8+ cells to perform tumour cytotoxic functions. Abstract previously presented? yes (NCRI Liverpool) Any disclosures? no ()
P14. Bridging the gap between bench and bedside: Systematic review of treatment in experimental acute pancreatitis M.A. Javed 1,2, K. Altaf 1, H. Wei 1, D.N. Criddle 1,2, R. Sutton 1. 1
Liverpool NIHR Pancreas Biomedical Research Unit, Royal Liverpool University Hospital, UK 2 Department of Cellular and Molecular Physiology, University of Liverpool, Liverpool, UK
Category: Benign and Inflammatory Introduction: Experimental animal models of acute pancreatitis (AP) are well characterised, have been used to investigate disease mechanisms and test potential therapies before human trials. Objective: To evaluate current evidence for the treatment of experimental AP and assess the translation of these preclinical studies into randomised clinical trials (RCTs) in patients with AP. Methods: A comprehensive online search of medline, embase, Pubmed and the Cochrane library was conducted by two independent reviewers of all published preclinical studies and corresponding RCTs of AP treatment undertaken from the first reported studies to January 2012. Results: 142 compounds have been tested in 257 experimental studies. Of these, only 21 compounds have been tested in 116 RCTs. modes of action included microcirculatory modification [Trials (T) 38, Compounds (C) 19; RCTs 4, multiple compounds in individual RCTs (CRCT) 2], anti-inflammatory agents (T 31, C 21, RCTs 0), eicosanoid modulators (T 32, C 16; RCTs 3, CRCT 1), immune-modulators (T 35, C 23; RCTs 13, CRCT 5), protease inhibitors (T 31, C 12; RCTs 34, CRCT 5), secretion inhibitors (T 28, C 13; RCTs 40, CRCT 2), antioxidants (T 18, C 8; RCTs 14, CRCT 4) and compounds with multiple actions (T 24, C 16; RCTs 6, CRCT 2). Conclusion: While attrition is expected in drug development, our findings identify a significant translational gap between animal studies and RCTs in AP. We propose standardised reporting of preclinical studies to improve the quality, comparability and translation of experimental AP treatment. Take-home message – There is a lot of potential for improving translational research in acute pancreatitis in order to bridge the gap between bench and bedside. Abstract previously presented? yes (EPC meeting) Any disclosures? no ()
P15. Nuclear translocation of FGFR1 and FGF2 in pancreatic stellate cells is necessary for pancreatic cancer cell invasion Stacey J. Coleman, Athina-Myrto Chioni, Mohammed Ghallab, Rhys K. Anderson, Nicholas R. Lemoine, Richard P. Grose, Hemant M. Kocher. Barts Cancer Institue, University of Queen Mary, UK Category: Malignant Background: In pancreatic cancer, a number of FGFs and their cognate receptors are over-expressed and linked to poor prognosis. How FGF ligands and receptors interact within pancreatic stellate cells (PSCs), to drive pancreatic cancer progression, is unclear. We investigated whether nuclear translocation of FGFR1 with FGF2 plays a role in PSC behaviour. Design: FGF2 and FGFR1 RNAi, together with an FGFR inhibitor (PD173074) were used in pancreatic cancer cell lines, PSCs and in organotypic cultures. Localisation of FGFR1 and FGF2 in the nucleus was also studied in vivo. Results: FGF2 and FGFR1 localised to the nucleus in stromal fibroblasts of human pancreatic cancer tissue. FGFR1 and FGF2 co-localised to the nucleus in PSCs but not in cancer cell lines. Abolishing FGFR1 and FGF2 functions using either RNAi or FGFR inhibitor in PSCs correlated with G1 cell-cycle block and a significant reduction in cell proliferation. In an organotypic model, nuclear FGFR1 and FGF2 were significantly greater in PSCs invading into the matrix. Furthermore, effective blockade of nuclear FGFR1 signalling (using an FGFR inhibitor) in PSCs abolished cancer cell invasion. Conclusion: These studies show for the first time that nuclear FGFR1 and FGF2 may play a role in driving pancreatic stellate cell proliferation. Preventing nuclear FGF/FGFR mediated proliferation in PSCs leads to disruption of the tumour microenvironment, thus preventing pancreatic cancer cell invasion.
Abstracts / Pancreatology 13 (2013) e1–e20
vs. 79.1%). In HC, CP and PC, the combination of three markers (iC3b, ULBP2 and CA 19.9) has better predictive value than CA19.9 alone or combination of ULBP2 and CA19.9 or combination of iC3b and CA19.9 (79.6% vs. 78% vs. 77.8% vs. 79.4%). Conclusion: Our preliminary results show that there is possibility of better predictive value in sensitivity and specificity with a combination of markers rather than a single marker alone. More samples are needed to qualify this finding. Take-home message: A combination of markers may have better predictive value than a single marker alone. Abstract previously presented? no () Any disclosures? no ()
P13. Activated pancreatic stellate cells inhibit CD8+ infiltrate in pancreatic ductal adenocarcinoma via CXCL12 Abasi Ene-Obong, Andrew J. Clear, Alan G. Ramsay, Hemant M. Kocher. Barts Cancer Institute, UK Category: Malignant Background: Pancreatic ductal adenocarcinoma (PDAC) is characterised by a prominent immune cell response. We hypothesized a differential immune cell infiltrate in distinct stromal compartments. Method: Specific tissue microarrays focusing on juxta-tumoural stroma (<100 mm from tumour) from panstroma from various pancreatico-biliary diseases were stained for distinct immune cell markers. Immune cell densities of the stromal compartments were carried out using automated software. Prognostic significance was determined with X-Tile. In vitro and in vivo assays were performed to delineate the mechanistic aspect of the immune infiltrate pattern. Results: PDAC had more immune cell infiltrate when compared to other pancreatico-biliary diseases except for CD8+, which was lower and comparable to that of chronic inflammation (p<0.001). CD3+, CD8+, Foxp3+, CD20+ cells PDAC could not infiltrate the juxta-tumoural stromal compartment whereas myeloperoxidase+, CD4+, CD68+ could do so, in two cohorts (p<0.05). PDAC patients with higher CD8+ densities survived longer than patients with lower densities (p ¼0.03). KPC mice stroma demonstrated similar juxta-tumoral exclusion which could be reversed upon stromal collapse by targeting stellate cells (p<0.05). In vitro migration assays demonstrated increased T-cell migration towards activated pancreatic stellate cells (PSC) compared to quiescent PSC (p¼0.0006), which was abrogated by knockdown of CXCL12 in aPSC (p<0.01). Adhesion assays demonstrated increased adhesion of T-cells to aPSC than qPSC (p¼0.0079). Conclusion: Activated stellate cells hinder the migration of T-cells, specifically, cytotoxic CD8+ T-cells, towards the juxta-tumoural stroma due to secreted CXCL12. Rendering aPSC quiescent reduces CXCL12 production which may enable CD8+ cells to perform tumour cytotoxic functions. Take-home message: Activated pancreatic stellate cells hinder cytotoxic CD8+ T cell infiltrate to the tumour. Rendering activated pancreatic stellate cells quiescent may enable CD8+ cells to perform tumour cytotoxic functions. Abstract previously presented? yes (NCRI Liverpool) Any disclosures? no ()
P14. Bridging the gap between bench and bedside: Systematic review of treatment in experimental acute pancreatitis M.A. Javed 1,2, K. Altaf 1, H. Wei 1, D.N. Criddle 1,2, R. Sutton 1. 1
Liverpool NIHR Pancreas Biomedical Research Unit, Royal Liverpool University Hospital, UK 2 Department of Cellular and Molecular Physiology, University of Liverpool, Liverpool, UK
Category: Benign and Inflammatory Introduction: Experimental animal models of acute pancreatitis (AP) are well characterised, have been used to investigate disease mechanisms and test potential therapies before human trials. Objective: To evaluate current evidence for the treatment of experimental AP and assess the translation of these preclinical studies into randomised clinical trials (RCTs) in patients with AP. Methods: A comprehensive online search of medline, embase, Pubmed and the Cochrane library was conducted by two independent reviewers of all published preclinical studies and corresponding RCTs of AP treatment undertaken from the first reported studies to January 2012. Results: 142 compounds have been tested in 257 experimental studies. Of these, only 21 compounds have been tested in 116 RCTs. modes of action included microcirculatory modification [Trials (T) 38, Compounds (C) 19; RCTs 4, multiple compounds in individual RCTs (CRCT) 2], anti-inflammatory agents (T 31, C 21, RCTs 0), eicosanoid modulators (T 32, C 16; RCTs 3, CRCT 1), immune-modulators (T 35, C 23; RCTs 13, CRCT 5), protease inhibitors (T 31, C 12; RCTs 34, CRCT 5), secretion inhibitors (T 28, C 13; RCTs 40, CRCT 2), antioxidants (T 18, C 8; RCTs 14, CRCT 4) and compounds with multiple actions (T 24, C 16; RCTs 6, CRCT 2). Conclusion: While attrition is expected in drug development, our findings identify a significant translational gap between animal studies and RCTs in AP. We propose standardised reporting of preclinical studies to improve the quality, comparability and translation of experimental AP treatment. Take-home message – There is a lot of potential for improving translational research in acute pancreatitis in order to bridge the gap between bench and bedside. Abstract previously presented? yes (EPC meeting) Any disclosures? no ()
P15. Nuclear translocation of FGFR1 and FGF2 in pancreatic stellate cells is necessary for pancreatic cancer cell invasion Stacey J. Coleman, Athina-Myrto Chioni, Mohammed Ghallab, Rhys K. Anderson, Nicholas R. Lemoine, Richard P. Grose, Hemant M. Kocher. Barts Cancer Institue, University of Queen Mary, UK Category: Malignant Background: In pancreatic cancer, a number of FGFs and their cognate receptors are over-expressed and linked to poor prognosis. How FGF ligands and receptors interact within pancreatic stellate cells (PSCs), to drive pancreatic cancer progression, is unclear. We investigated whether nuclear translocation of FGFR1 with FGF2 plays a role in PSC behaviour. Design: FGF2 and FGFR1 RNAi, together with an FGFR inhibitor (PD173074) were used in pancreatic cancer cell lines, PSCs and in organotypic cultures. Localisation of FGFR1 and FGF2 in the nucleus was also studied in vivo. Results: FGF2 and FGFR1 localised to the nucleus in stromal fibroblasts of human pancreatic cancer tissue. FGFR1 and FGF2 co-localised to the nucleus in PSCs but not in cancer cell lines. Abolishing FGFR1 and FGF2 functions using either RNAi or FGFR inhibitor in PSCs correlated with G1 cell-cycle block and a significant reduction in cell proliferation. In an organotypic model, nuclear FGFR1 and FGF2 were significantly greater in PSCs invading into the matrix. Furthermore, effective blockade of nuclear FGFR1 signalling (using an FGFR inhibitor) in PSCs abolished cancer cell invasion. Conclusion: These studies show for the first time that nuclear FGFR1 and FGF2 may play a role in driving pancreatic stellate cell proliferation. Preventing nuclear FGF/FGFR mediated proliferation in PSCs leads to disruption of the tumour microenvironment, thus preventing pancreatic cancer cell invasion.