- Email: [email protected]
1114 TRANSPLANTATION OF EpCAM+ve HUMAN HEPATIC STEM CELLS IN LIVER CIRRHOSIS AND CELLULAR IMMUNE RESPONSE
POSTERS 1114 TRANSPLANTATION OF EpCAM+ve HUMAN HEPATIC STEM CELLS IN LIVER CIRRHOSIS AND CELLULAR IMMUNE RESPONSE M.A. Habeeb1 , A.A. Khan2 , A. Bardia2 , G. Sivaram2 , G. Pande3 . 1 Gastroenterology and Hepatology, 2 Centre for Liver Research and Diagnostics, Deccan College of Medical Sciences, 3 Centre for Cellular and Molecular Biology, Hyderabad, India E-mail: [email protected] Introduction: Liver transplantation is the only effective treatment for decompensated liver-cirrhosis. Several factors, such as nonavailability of donors, operative-risks, complications associated with rejection, usage of immunosuppressive agents, and high cost of treatment, make this strategy available to only a few people. Human foetal liver derived hepatic progenitor cell transplantation (HSCT) have shown to be a good alternative to manage end-stage liver diseases. In this retrospective study, we investigated safety and efficacy of HSCT by monitoring the T-cell, NK-cell and cytokines which play major role in cellular immune response and rejection of chronic decompensated liver cirrhosis patients. Materials and Methods: A total of 5 patients with decompensated liver cirrhosis were enrolled in the study. After giving human foetal liver-derived EpCAM positive cell transplantation, T-cell (CD3, CD4 and CD8), NK-cells (CD16) by flowcytometry and cytokine-levels (IL2, TNFb, IFNa, IFNb and IFNg) by ELISA were monitored four times within a month. Result: Present study demonstrated that after HSCT patient showed marked clinical recovery and decline in the MELD score and there was no significant variation found in cell mediated response and cytokine levels between pre and post transplantation. Conclusion: Hence this preliminary study demonstrated human foetal liver-derived EpCAM positive stem cell transplantation is safe for end stage liver cirrhosis. 1115 IDENTIFYING COREGULATED GENE CLUSTERS LINKED TO HEPATIC FIBROSIS R. Hall1 , R. Liebe1 , K. Hochrath1 , S.N. Weber1 , R. Alberts2 , K. Schughart2 , R.W. Williams3 , F. Lammert1 . 1 Department of Medicine II, Saarland University Medical Center, Homburg, 2 Department of Experimental Mouse Genetics, Helmholtz Centre for Infection Research, Braunschweig, Germany; 3 Department of Anatomy and Neurobiology, University of Tennessee, Memphis, TN, USA E-mail: [email protected] Background and Aims: Liver fibrosis is a complex trait that varies considerably among individuals. The underlying networks of genetic factors have not yet been clarified. Quantitative trait loci (QTL) analysis allows the identification of genetic loci, interacting genes and gene networks associated with fibrosis progression. Here, we provide the first systematic transcriptome analysis of liver fibrosis in a ‘genetic reference panel’ of recombinant inbred (RI) lines differing in fibrosis susceptibility. The aim of our study was to identify genetic networks and regulatory mechanisms of gene expression during fibrogenesis. Methods: We generated 96 hepatic expression profiles of fibrotic livers, using Mouse Gene 1.0 ST microarrays (Affymetrix), from 32 RI BXD lines after fibrosis induction by carbon tetrachloride (CCl4 ) challenge. Transcript levels were used as traits in QTL analysis and mapped to the BXD mouse genomes. Thereby, we identified loci regulating gene expression during fibrogenesis (eQTL). We defined selection criteria for candidate genes, within the hydroxyproline- and fibrosis-stage associated regions (pQTL) identified previously. These implicated the search for genes that 1) are locally regulated quantitative trait genes (cisQTG), 2) cosegregate significantly with fibrosis phenotypes and 3) show a fibrosis-specific regulation by comparing cisQTG in CCl4 -treated fibrotic animals to expression data of healthy animals.
Results: Among over 1000 genes in pQTL regions, this eQTL mapping strategy identified 61 cisQTG; 35 cisQTG were differentially regulated compared to healthy animals, including known fibrosisassociated genes such as nuclear receptor LXR and tenascin C. The significant correlation of the fibrosis specific cisQTG to liver phenotypes and the presence of non-synonymous single nucleotide polymorphisms within coding regions of the cisQTG confirmed their relevance for fibrogenesis. Finally, we describe unique networks of the identified candidate genes co-regulated during fibrogenesis. Conclusions: Integrating QTL mapping of expression and phenotype data within the BXD RI panel as a ‘genetic reference population’ allowed us to identify novel candidate genes of liver fibrogenesis. Our experimental set-up provides a basic experimental framework to dissect gene networks that drive hepatic fibrogenesis and are modulated by therapeutic interventions. 1116 THE CHEMOKINE MACROPHAGE INFLAMMATORY PROTEIN-1a (MIP-1a) IS AN IMPORTANT MEDIATOR OF LIVER FIBROSIS D. Heinrichs, M.-L. Berres, A. Nellen, P. Fischer, C. Trautwein, H.E. Wasmuth, H. Sahin. University Hospital Aachen, Aachen, Germany E-mail: [email protected] Background and Aims: Chemokines are important mediators of acute and chronic liver injury. Overall, chronic liver fibrosis is strongly linked to activation of hepatic stellate cells as well as to influx of inflammatory cells into the liver. We here investigate the role of the chemokine CCL3, also known as macrophage inflammatory protein-1a, in two experimental liver fibrosis models. Methods: CCL3−/− mice and wild-type mice were treated with CCl4 or a MCD diet to induce liver fibrosis. Fibrosis was analyzed by Sirius red staining, hydroxyproline content and gene expression of fibrosis-associated genes. Immune cell infiltration was investigated by FACS analysis and immunohistochemical staining. The proliferation of the stellate cell line GRX was determined after stimulation with recombinant CCL3, in vitro. Results: We could show that the protein concentration of CCL3 is increased in wild-type mice after chronic liver injury (P < 0.05). CCL3 deficient mice showed significant decreased levels of fibrosis as assessed by histological staining and intrahepatic hydroxyproline content in both fibrosis models (P < 0.05). The activation of stellate cells was decreased in the CCL3−/− mice in contrast to the wildtype counterparts. FACS analysis and immunohistochemical staining showed a significant decrease of the T-cell population in the liver (P < 0.05). Stimulation of stellate cells with recombinant CCL3 led to significantly increased proliferation when compared to vehicle treated cells (P < 0.001). Conclusions: Our results identify the chemokine CCL3 as an important mediator of liver fibrogenesis. The inhibition of fibrosis in CCL3−/− mice in different fibrosis models is triggered by decreased activation of hepatic stellate cells and influx of recruited T-cells into the liver. Thus, therapeutical modulation of CCL3 might be a promising target for chronic liver diseases.
Journal of Hepatology 2013 vol. 58 | S409–S566
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Results: Among over 1000 genes in pQTL regions, this eQTL mapping strategy identified 61 cisQTG; 35 cisQTG were differentially regulated compared to healthy animals, including known fibrosisassociated genes such as nuclear receptor LXR and tenascin C. The significant correlation of the fibrosis specific cisQTG to liver phenotypes and the presence of non-synonymous single nucleotide polymorphisms within coding regions of the cisQTG confirmed their relevance for fibrogenesis. Finally, we describe unique networks of the identified candidate genes co-regulated during fibrogenesis. Conclusions: Integrating QTL mapping of expression and phenotype data within the BXD RI panel as a ‘genetic reference population’ allowed us to identify novel candidate genes of liver fibrogenesis. Our experimental set-up provides a basic experimental framework to dissect gene networks that drive hepatic fibrogenesis and are modulated by therapeutic interventions. 1116 THE CHEMOKINE MACROPHAGE INFLAMMATORY PROTEIN-1a (MIP-1a) IS AN IMPORTANT MEDIATOR OF LIVER FIBROSIS D. Heinrichs, M.-L. Berres, A. Nellen, P. Fischer, C. Trautwein, H.E. Wasmuth, H. Sahin. University Hospital Aachen, Aachen, Germany E-mail: [email protected] Background and Aims: Chemokines are important mediators of acute and chronic liver injury. Overall, chronic liver fibrosis is strongly linked to activation of hepatic stellate cells as well as to influx of inflammatory cells into the liver. We here investigate the role of the chemokine CCL3, also known as macrophage inflammatory protein-1a, in two experimental liver fibrosis models. Methods: CCL3−/− mice and wild-type mice were treated with CCl4 or a MCD diet to induce liver fibrosis. Fibrosis was analyzed by Sirius red staining, hydroxyproline content and gene expression of fibrosis-associated genes. Immune cell infiltration was investigated by FACS analysis and immunohistochemical staining. The proliferation of the stellate cell line GRX was determined after stimulation with recombinant CCL3, in vitro. Results: We could show that the protein concentration of CCL3 is increased in wild-type mice after chronic liver injury (P < 0.05). CCL3 deficient mice showed significant decreased levels of fibrosis as assessed by histological staining and intrahepatic hydroxyproline content in both fibrosis models (P < 0.05). The activation of stellate cells was decreased in the CCL3−/− mice in contrast to the wildtype counterparts. FACS analysis and immunohistochemical staining showed a significant decrease of the T-cell population in the liver (P < 0.05). Stimulation of stellate cells with recombinant CCL3 led to significantly increased proliferation when compared to vehicle treated cells (P < 0.001). Conclusions: Our results identify the chemokine CCL3 as an important mediator of liver fibrogenesis. The inhibition of fibrosis in CCL3−/− mice in different fibrosis models is triggered by decreased activation of hepatic stellate cells and influx of recruited T-cells into the liver. Thus, therapeutical modulation of CCL3 might be a promising target for chronic liver diseases.
Journal of Hepatology 2013 vol. 58 | S409–S566
S455