脂肪肝最新研究摘要EASL.doc

PS066FXR AGONISM BY GS-9674 DECREASES STEATOSIS AND FIBROSIS INA MURINE MODEL OF NASHJ.T. Liles1, S. Karnik1, E. Hambruch2, C. Kremoser2, M. Birkel2, W.J. Watkins1, D. Tumas1, D. Breckenridge1, D. French1. 1Gilead Sciences,Foster City, United States; 2Phenex Pharmaceuticals AG, Heidelberg,GermanyE-mail: John.LilesGBackground and Aims: GS-9674 is a selective nonsteroidal FarnesoidX Receptor (FXR) agonist being developed by Gilead Sciences for thetreatment of NASH. FXR is a nuclear hormone receptor that is highlyexpressed in the gastrointestinal tract and liver, and is a primaryregulator of bile acid (BA) homeostasis. GS-9674 acts in the intestineto produce FGF19 (FGF15 in rodents), an endocrine hormone thatdown-regulates BA synthesis, lipogenesis and gluconeogenesis in theliver and increases energy expenditure in adipose tissue. Both lowFGF19 and high BA levels have been associated with the pathogenesisand progression of NASH including the degree of hepatic fibrosis. Inthis study we evaluated the therapeutic efficacy of GS-9674 in micewith established NASH.Methods:NASHwas induced in male C57BL/6 mice byadministrationof a fast food diet (FFD) high in fat, cholesterol, and sugar for 240 days.Animals were subsequently treated with either vehicle or GS-9674(30 mg/kg, BID, PO) for 90 days (n = 15/group). Endpoints includedsteatosis evaluated by morphometry, liver hydroxyproline levels,clinical pathology, NAFLD activity scores (NAS) by histology, and anoral glucose tolerance test. C57BL/6 mice on a normal diet (n = 15/group) were a control group.Results: Serum cholesterol, AST and ALT, and liver steatosis andfibrosis were significantly elevated in vehicle-treated mice following240 and 330 days of FFD relative to control mice. Compared tovehicle-treated mice, mice treated with GS-9674 from day 240 to 330demonstrated significant decreases in serum AST (192 21 vs. 293 12 IU/L, p 0.0002), ALT (178 35 vs. 267 24 IU/L, p 0.04) andcholesterol (144 12 vs. 321 23 mg/dl, p 0.0001). GS-9674treatment also significantly reduced hepatic steatosis as measuredby morphometric analysis (7.4 1.3 vs. 14.5 0.7% area, p 0.01). GS-9674-treatment reduced liver fibrosis as measured by hydroxyprolinecontent (127 12 vs. 320 37 mmol/g, p 0.0001) compared tovehicle-treated animals. GS-9674 did not affect food intake, bodyweight, or glucose metabolism.Conclusions: In a murine model of NASH, treatment with the FXRagonist GS-9674 significantly reduced serum ALTand ASTand hepaticsteatosis and fibrosis. The data support clinical evaluation of GS-9674for the treatment of NASH.PS067IDENTIFICATION OF LIPIDOMIC SIGNATURES THAT DEFINE THREESPECIFIC SUBTYPES OF NAFLD AND DIFFERENTIATE NASH FROMSIMPLE STEATOSISC. Alonso1, D. Fernandez-Ramos2, M. Iruarrizaga-Lejarreta1,M. Varela2, I. Martinez-Arranz1, M. Noureddin3, M.L. Martinez-Chantar2, S.C. Lu3, P. Ortiz1, J.M. Mato2. 1OWL; 2CICbioGUNE, Ciberehd, Derio, Spain; 3Division of Gastroenterology, Cedars-Sinai Medical Center, Los Angeles, United StatesE-mail: jmmatocicbiogune.esBackground and Aims: Nonalcoholic steatohepatitis (NASH) is ahistological definition that groups together defects in diversebiochemical processes causing hepatic fat accumulation,inflammation, necrosis and fibrosis. The identification of the typesof mechanisms leading to NASH and the discovery of noninvasivebiomarkers of NASH subtypes are central for the development ofeffective treatments and precise diagnosis. This study aims to capturethe metabolic architecture of the main NASH subtypes to help defineeffective treatments and discover specific serum metabolomicpatterns reflective of each NASH subtype.Methods: We have compared the serum metabolome (over 400different molecular species) of a mouse model (Mat1a-/-) thatspontaneously develops NASH with that of WT mice and selectedthe top fifty metabolites that more significantly differentiatedbetween both genotypes (p 1E-05).Results: Silhouette cluster analysis revealed that this metabolicsignature sub-classified a cohort of 377 patients with biopsy provenNAFLD (246 diagnosed of steatosis and 131 diagnosed of NASH) intothree clusters: a first cluster (n = 116) showing a serum metabolicprofile similar to that observed in the Mat1a-/- mice (M-subtype),a second cluster (n = 115) showing the opposite metabolomicprofile (non-M-subtype) and a third cluster (n = 146) presenting anintermediate metabolic profile (I-subtype). Next, we wonderedwhether NAFLD patients in the M-type NAFLD subgroup could befurther separated into simple steatosis and NASH based exclusively intheir metabolic profile. Volcano plot analysis representation oflog10(p-value) and log2(fold-change) identified a group of highlysignificant lipids (p 1E-05, mostly lysophospholipids andpolyunsaturated fatty acids) that effectively differentiated betweenthese two conditions. Similarly,we successfully identified a metabolicsignature (mostly triglycerides and phosphatidylcholines) thataccurately differentiated between steatosis and NASH in the non-MtypeNAFLDsubgroup.Whenthis unsupervised approachwas appliedto the third cluster (I-subtype), the metabolic signature able todifferentiate between steatosis and NASH was mainly based inpolyunsaturated fatty acids.Conclusions: Our data document the power of serum metabolomicsto identify signatures that define NAFLD subtypes and differentiateNASH from simple steatosis in humans. Moreover, these metabolicsignatures may be used to analyze the individual response totreatment in clinical trials.PS068THE NEW GENERATION PAN-PPAR AGONIST IVA337 PROTECTS THELIVER FROM METABOLIC DISORDERS AND FIBROSISG. Wettstein1, C. Estivalet1, J. Tessier1, P.T. Boustugue1, I. Jantzen1,E. Defrene1, F. Kupkowski1, P. Faye1, V. Adarbes1, J.-M. Germain1,E. Vasseur1, C. Robert1, C. Fromond1, P. Broqua1, J.-L. Junien1,J.-M. Luccarini1, I. Konstantinova1. 1INVENTIVA, daix, FranceE-mail: Background and Aims: Non-alcoholic fatty liver disease (NAFLD) isa complex liver pathology starting from simple hepatocellularsteatosis to non-alcoholic steatohepatitis (NASH), fibrosis andultimately cirrhosis. NASH is proposed as the hepatic manifestationof the metabolic syndrome. Currently there is no treatment forNASH although several Peroxisome Proliferator-Activated Receptors(PPARs) agonists have demonstrated positive signals during clinicaltrials in NASH. PPARs play roles in lipid and glucose metabolism,inflammation, cellular growth, differentiation and fibrogenesis. Thissuggests that, IVA337, a well-balanced pan-PPAR agonist (EC50:PPAR 0.9 M, PPAR 0.5 M, PPAR 0.2 M), that underwent Phase 2clinical investigations in Type 2 diabetic patients, might represent anattractive therapeutic approach in NASH. We evaluated the effect ofIVA337 in vitro on proliferation and activation of Hepatic Stellate Cells(HSC) and in vivo on key parameters of the metabolic syndrome aswell as on hepatic fibrosis.Methods: In vitro we studied the effects of IVA337 on PDGF-inducedproliferation and stiffness-induced activation of Hepatic Stellate Cells.In vivo, we investigated IVA337 in models of High Fat/High Sucrose(HF/HS) diet and CCL4 induced liver fibrosis (prophylactic andtherapeutic).Results: Treatment with IVA337 but not with selective PPAR agonistsled to a complete dose-dependent inhibition of PDGF-inducedproliferation in HSC. IVA337 also inhibited HSC activation bypreventing upregulation of -SMA expression. In the HF/HS dietmodel, IVA337 given upon therapeutic dosing regimen for 4 weeks,resulted in a dose-dependent decrease of body weight, serumtriglycerides, adiposity index, insulin resistance and an increase ofserum adiponectin. IVA337 also inhibited CCL4-induced fibrosis(prophylactic and therapeutic), expression of TGF- familymembers and extracellular matrix components.Conclusions: These findings demonstrate that simultaneousactivation of the 3 PPAR isoforms by IVA337 exerts a beneficialeffect on HSC proliferation and differentiation in vitro, and on liverfibrosis in vivo. Interestingly, selective PPARs agonists did not inhibitHSC proliferation, pointing out the benefit of using a pan-PPARagonist for the treatment of liver fibrosis. In addition, IVA337demonstrated positive effects on metabolic markers consideredroot causes for NASH. Together this data support the clinicalinvestigation of IVA337 for the treatment of NASH patients.PS069THE EFFECT OF MTORC1 INHIBITION ON NAFLD AND SUBSEQUENTHCC DEVELOPMENTA. Umemura1,2, Y. Itoh1, M. Karin2. 1Department of MolecularGastroenterology and Hepatology, Graduate School of Medical Science,Kyoto Prefectural University of Medicine, Kyoto, Japan; 2Laboratory ofGene Regulation and Signal Transduction, Departments of Pharmacologyand Pathology, UCSD, San Diego, United StatesE-mail: aumemura2002yahoo.co.jpBackground and Aims: Since mTORC1 is activated in up to 50% ofHCCs, there has been much interest in the use of mTORC1 inhibitorsfor HCC treatment. mTORC1 is also activated in response to obesitywhich greatly enhances HCC risk. So, we hypothesized that mTORC1inhibition attenuates obesity-induced fatty liver and its subsequentHCC development.Methods: We conducted short-term mTORC1 inhibition by amTORC1 inhibitor rapamycin (A). To determine the direct effect oflong-term mTORC1 suppression in hepatocytes,we used hepatocytespecificRaptor-deficient (Rpt KO) mice (B). We also used Tsc1knockout (TSC1 KO) mice, in which mTORC1 is constitutivelyactivated (C).Results: (A) Rapamycin treatment improved fatty liver in HFD (highfatdiet)-fed mice, surprisingly however, such mTORC1 inhibitionalso resulted in increased IL-6 production, and activation of STAT3,which is previously shown to enhance HCC development. (B)Unexpectedly, complete mTORC1 inhibition by Raptor ablation inhepatocytes resulted in not only increased IL-6 production andSTAT3 activation, but also strongly potentiating DEN-inducedHCC development in HFD-induced obese mice. Thus, mTORC1activation does not play a crucial role in obesity-promoted HCCdevelopment, at least in this model. (C) On the other hand, it isreported that mTORC1 inhibition prevented spontaneous liverdamage, fibrosis and tumorigenesis in TSC1KO mice, in whichmTORC1 is hyperactivated. We then generated TSC1/ p62 doubleknock-out mice to investigate the role of p62, a multifunctionalprotein in TSC1 KO liver. In the absence of p62, liver damage andfibrosis observed in TSC1 KO mice were attenuated, and mostimportantly, the tumorigenesis was completely abolished.Conclusions: mTORC1 inhibition suppresses liver damage, fibrosisand tumorigenesis in a mTORC1-hyperactive state. However,in normal mice, even obese ones, the adverse effects of mTORC1inhibition outweigh its benefits. Recently, increased liver damagewasalso observed during a clinical trial of a mTOR inhibitor everolimus(EVOLVE-1), and this trial did not meet the primary end point.Hyperactivation of AKT and aberrant liver regeneration, also seenafter rapamycin treatment and in Rpt KO livers, may be reasons to beconcerned about long-term mTORC1 inhibition as an HCCtreatment. Of note that patients with HCC usually underlie achronic liver disease, typically liver cirrhosis and requireregeneration to maintain their liver functions. In conclusion,mTORC1 inhibition may not be a suitable treatment for HCC.PS070REDUCTION OF LIVER STEATOSIS AND FIBROSIS WITH AN ASK1INHIBITOR IN A MURINE MODEL OF NASH IS ACCOMPANIED BYIMPROVEMENTS IN CHOLESTEROL, BILE ACID AND LIPIDMETABOLISMG. Budas1, S. Karnik1, T. Jonnson2, T. Shafizadeh2, S. Watkins2,D. Breckenridge1, D. Tumas1. 1Gilead Sciences, Foster City; 2Metabolon,Durham, United StatesE-mail: Background and Aims: GS-4997 is a first-in-class, small moleculeinhibitor of ASK1 that is in clinical development for the treatment ofNASH. ASK1 is a redox-sensitive kinase which is activated by variouspathological stimuli including oxidative stress, TGF-b andhyperglycemia. Activation of ASK1 mediates activation of JNK andp38 MAP kinases which can induce hepatocyte apoptosis, hepaticinflammation and myofibroblast activation leading to fibrosis. Wepreviously demonstrated that ASK1 inhibition reduces hepaticsteatosis and fibrosis and improves insulin sensitivity in preclinicalmodels of NASH. In the current study, we comprehensivelycharacterized the effect of ASK1 inhibition on the metabolome in amurine model of NASH.Methods: Male C57BL/6 mice were fed a diet high in fat, cholesterol,and sugar (FF diet) for 330 days. A NASH phenotype wasestablished by day 240, after which animals were treated for 90days with a small molecule inhibitor of ASK1 (GS-444217) or withvehicle (n = 15/group). Animals fed a normal dietwere used as controlgroup. Endpoints included hepatic steatosis evaluated bymorphometry, clinical pathology, liver hydroxyproline levels, anoral glucose tolerance test and mass spectrometric analysis of plasmaand liver using the MetabolonO platform.Results: Chronic administration of the FF diet caused liver steatosis,fibrosis and insulin resistance, all of whichwere significantly reducedby treatment with an ASK1 inhibitor. The FF diet also significantlyincreased hepatic and serum cholesterol and serum bile acids. ASK1inhibition reduced serum and liver cholesterol by 14% and 45%,respectively, and strongly reduced serum bile acids including cholate(91% reduction) and deoxycholate (90% reduction). FF diet inducedalterations in hepatic fatty acid metabolism similar to that observedin human NASH. These alterations included increased hepatic levelsof monounsaturated fatty acids produced by stearoyl CoA desaturases(palmitoleic and oleic acids) and increased total hepaticdiacylglycerol and triacylglycerol levels. These FF diet-inducedalterations to the hepatic lipid profile were significantly reduced byASK1 inhibition.Conclusions: Inhibition of ASK1 in a murine model of NASH reducedhepatic steatosis and fibrosis. This efficacy was also accompanied byimprovements in cholesterol, bile acid and lipid metabolism asdetermined by unbiased metabolic profiling.PS071HSP72 OVEREXPRESSION PROTECTS FROM LIVER INJURY VIAATTENUATION OF JNK SIGNALLINGK. Levada1, N. Guldiken1, G. Vella1, L.P. James2, J. Haybaeck3, A.K. Kiemer4, S.M. Kessler4, C. Trautwein1, P. Strnad1. 1Department ofMedicine III and IZKF, University Hospital Aachen, Aachen, Germany;2Department of Pediatrics, Arkansas Childrens Hospital ResearchInstitute, Little Rock, United States; 3Institute of Pathology, MedicalUniversity of Graz, Graz, Austria; 4Department of Pharmacy,Pharmaceutical Biology, Saarland University, Saarbrcken, GermanyE-mail: Background and Aims: Heat shock protein (Hsp) 72 is a molecularchaperone that is upregulated in response to a variety of stresssituations and possesses broad cytoprotective functions. To elucidateits hepatic function, we studied its expression in various humanliver disorders and its biological significance in newly generatedtransgenic animals.ORAL PRESENTATIONSS170 Journal of Hepatology 2016 vol. 64 | S159S182Methods: Transgenic mice overexpressing Hsp72 under the controlof a tissue-specific tetracycline-inducible system were crossed withanimals carrying the tetracycline-responsive transactivator under thecontrol of the liver activator protein promoter (Hsp72-LAP mice).Acute liver injury was induced by a single intraperitoneal injection ofacetaminophen (800 mg/kg), whereas 8 week feeding withmethionine choline-deficient diet (MCD) was used to inducelipotoxic liver damage. Primary hepatocytes were subjected totreatment with palmitic acid (0.5 mM palmitic acid for 24 h).Results: Patients with non-alcoholic steatohepatitis and chronichepatitis C infection displayed significantly elevated Hsp72 mRNAlevels. Moreover, Hsp72 levels progressively increased with thehigher extent of hepatic inflammation. Hsp72-LAP mice displayeddoxycycline-regulated, robust Hsp72 overexpression in hepatocytes,but not in the other tissues or cell types. Primary hepatocytesisolated from these animals were more resistant towards theisolation-induced stress as determined through lower ALT and LDHlevels in the supernatant. Eighteen hours after acetaminopheninjection, a significantly lower liver injury was noted in Hsp72-LAPmice in comparison to single transgenes (ALT: 933 vs. 1977, p 0.05). Overexpression of Hsp72 protected also protected fromformation of APAP protein adducts (p = 0.03). After MCD-feeding,Hsp72-LAP mice displayed lower ALT levels (105 vs. 225, p = 0.02)and lower RIP-3 activation. Overexpression of Hsp72 did not affectthe extent of steatosis after MCD treatment (in mice) and palmiticacid treatment (in primary hepatocytes). On the other hand, in allthree models (isolated hepatocytes, APAP/MCD-treated mice),Hsp72-LAP mice/hepatocytes displayed significantly attenuatedJNK activation.Conclusions: Our results suggest that Hsp72 overexpression protectsagainst liver injury via attenuation of JNK signaling.LBP527SILYMARIN IMPROVES FIBROSIS IN NON-ALCOHOLICSTEATOHEPATITISW.K. Chan1, N.R.N. Mustapha2, S. Mahadeva1. 1Department ofMedicine, Faculty of Medicine, University of Malaya, Kuala Lumpur;2Department of Pathology, Hospital Sultanah Bahiyah, Alor Setar,MalaysiaE-mail: POSTER PRESENTATIONSJournal of Hepatology 2016 vol. 64 | S213S424 S225Introduction: Silymarin