UK Genetics of Liver Disease Consortium – ‘UK GoLD’ 1st MRC – IMPC Mouse Network Meeting, MRC Harwell, January 2012 Representatives Dr Quentin M. Anstee, Newcastle University Prof Alastair Burt, Newcastle University Prof Jeremy Lambert, Dundee University

Background Liver disease is 5th most common cause of death in UK. Common liver diseases are result of the interaction between environmental factors and multiple genetic factors which determine susceptibility. ONS, 2009

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Consortium Aims To identify genes that confer susceptibility to risk behaviors and determine disease severity; To understand mechanisms by which genes & gene-environment interactions cause disease; To validate GWAS and candidate gene associations through translational research; To identify and exploit novel potential therapeutic targets. To translate genetic association into a mechanistic understanding of disease pathogenesis and onwards to the identification of novel therapeutic targets

Cross-Cutting Research Themes An initiator of liver disease risk Mechanisms of Addiction Behavior Inflammation and hepatocellular injury Mechanisms of Metabolic Liver Disease The final common pathway of liver injury Modifiers of Hepatic Fibrosis and Liver Regeneration

Consortium Members Mechanisms of Addiction Behavior An initiator of liver disease risk * Prof J. Lambert Neuropharmacology Dundee * Dr D. Belelli * Prof D. Stephens Experimental Psychology Sussex * Prof T. Smart UCL Prof S. Brown Genetics MRC Harwell *§ Dr Q.M. Anstee Clinician Scientist Newcastle *§ Prof H.C. Thomas Imperial § Prof W. Wisden Molecular Neuroscience * MRC ‘GABA’ Addiction Cluster § MRC ‘ICCAM’ Addiction Cluster

Consortium Members Mechanisms of Addiction Behavior An initiator of liver disease risk * Prof J. Lambert University of Dundee * Dr D. Belelli * Prof D. Stephens Sussex University * Prof T. Smart UCL Prof S. Brown MRC Harwell *§ Dr Q.M. Anstee Newcastle University *§ Prof H.C. Thomas Imperial College § Prof W. Wisden * MRC ‘GABA’ Addiction Cluster § MRC ‘ICCAM’ Addiction Cluster

ENU Mutagenesis: A Route to Gene Discovery ENU-induced mutations of GABRB1 gene leads to increased alcohol consumption, increased motivation for alcohol, and increased sensitivity to alcohol. Mutant mice show massive increases in tonic GABA currents in accumbens medium spiny neurones that are integral to reward and motivation, suggesting that decreasing tonic GABA currents may reduce alcohol intake In humans, allelic differences of the GABRB1 gene were associated with alcohol dependence.

Linking Genes to Behaviour: Towards Endophenotypes Variations in the gene encoding a2 subunits of GABA receptors are associated with cocaine and alcohol addiction in people. Deleting a2 genes in mice reduces GABA currents in accumbens medium spiny neurones by 30% and prevents cocaine’s effects on conditioned behaviours. Variations in the gene for a2 are associated with cocaine addiction in people a2bg2 WT a2-/- a4bd Conditioned reinforcement in people

Consortium Members Mechanisms of Metabolic Liver Disease Inflammation and hepatocellular injury * Dr Q.M. Anstee Newcastle University Prof A. Burt * Prof C.P. Day * Prof A. Daly * Prof D. Jones * Prof R. Goldin Imperial College * Prof M. Thursz * Prof S. Brown MRC Harwell * Prof R. Cox * Dr P. Potter Prof D. Adams University of Birmingham Dr P. Newsome Dr G. Alexander Cambridge University * MRC Harwell Liver ENU Research Programme

Research Areas Non-Alcoholic Fatty Liver Disease Progressive steatohepatitis and fibrosis associated with T2DM, obesity and the metabolic syndrome. Tissue oxidative stress response. Role of caspase activation in disease pathogenesis. Immune Mediated Liver Injury Primary Biliary Cirrhosis Leucocyte-endothelial interactions and effector cell trafficking. Drug Induced Liver Disease & Toxicity Idiosyncratic drug reactions Paracetamol toxicity

GWAS Activity ‘FLIP’ – Fatty Liver Inhibition of Progression FP7 funded European research programme (Anstee, Burt, Daly, Day) GWAS of biopsy-proven NAFLD Establish large prospective and retrospective cohorts with NAFLD, NASH and NAFLD-related HCC for translational study. ‘UK PBC Consortium’ Recently completed GWAS of PBC (Jones, Mells, Alexander) Established the largest PBC cohort in the world for study. ‘GenomALC’ – Genetics of Alcoholic Liver Disease International study, GWAS of ALD (Day) ‘DILIGEN’ - Drug Induced Liver Injury GWAS of Idiosyncratic DILI across a range of agents (Daly, Day)

Consortium Members Modifiers of Hepatic Fibrosis and Liver Regeneration The final common pathway of liver injury Prof A. Burt Newcastle University Prof D. Mann Dr Q.M. Anstee Prof M. Thursz Imperial College Prof R. Goldin Dr P. Newsome University of Birmingham Prof D. Adams

Translation from Association to Mechanism & Therapy Role of coagulation system activation in Liver fibrosis and stellate cell activation. Translational studies demonstrate that carriers of FvL mutation have accelerated fibrosis. This was confirmed in FvL mice and anticoagulation demonstrated to be an effected anti-fibrotic. On-going MRC Experimental medicine funded clinical trial (WAFT-C). C57BL/6 FvL H&E Anti-SMA C57BL/6 + Warfarin

Targeting the Renin-Angiotensin System (RAS) to Treat Fibrosis Hepatic stellate cells have a RAS that prevents apoptosis. Drugs that induce apoptosis will stimulate fibrosis reversion despite ongoing liver injury.

UK GoLD Consortium Activity within the IMPC Current GWAS and hypothesis-driven research by UK GoLD members will help to guide IMPC gene prioritisation. UK GoLD members will establish a working group of clinicians to review IMPC clinical biochemistry data for metabolic, hepatitic and cholestatic compound phenotypes. UK GoLD will institute secondary histological screens for liver disease in selected knockout animals. Two internationally recognised expert liver pathologists (Burt and Goldin) with extensive experience in murine histopathology. Members will input into the phenotype pipelines to assess utility of screens and help guide technical development of the pipeline. Members will integrate with IMPC to use mice in established basic research programmes as specific knockouts become available.

UK GoLD Consortium Activity within the IMPC Current GWAS and hypothesis-driven research by UK GoLD members will help to guide IMPC gene prioritisation. UK GoLD members will establish a working group of clinicians to review IMPC clinical biochemistry data for metabolic, hepatitic and cholestatic compound phenotypes. Members will input into the phenotype pipelines to assess utility of screens and help guide technical development of the pipeline.

UK GoLD Consortium Activity within the IMPC Members will integrate with IMPC to use mice in established basic research programmes as specific knockouts become available. Potential UK GoLD Secondary Screens include: Electrophysiology (in vivo and in vitro), Addiction/motivational behaviours, Response to environmental challenges: EtOH, HFD, ALIOS, DILI, Fibrosis models: MCD, CCl4, Thiocetamide, BDL, etc. UK GoLD will institute histological screens for liver disease in selected knockout animals. Two internationally recognised expert liver pathologists (Burt and Goldin) with extensive experience in murine histopathology. Bidirectional translational Studies.