Digestive and Liver Disease - AISF · Digestive and Liver Disease ... Carlo Catassi, Ancona, Italy Umberto Cillo, Padua, ... Medicine, University of Milano-Bicocca, Milan, Italy

An International Journal of Gastroenterology and Hepatology

Digestiveand Liver Disease

Abstracts of the 48th A.I.S.F. – Italian Association for the Study of the Liver – Annual Meeting 2015 Rome, 19-20 February 2015

Volume 47 Supplement 1 15 February 2015 ISSN 1590-8658

An International Journal of Gastroenterology and HepatologyVol. 47 Supplement 1 (2015)

Digestiveand Liver Disease

Official Journal of:Italian Society of GastroenterologyFrench Society of GastroenterologyItalian Association for the Study of the LiverItalian Society for Digestive EndoscopyItalian Association for the Study of the PancreasItalian Society for Pediatric Gastroenterology, Hepatology and NutritionItalian Association for Hospital Gastroenterologists and Digestive EndoscopistsItalian Group for the Study of Infl ammatory Bowel Disease

SECTION EDITORS

Infl ammatory Bowel DiseaseLaurent Peyrin-Biroulet, Vandoeuvre, France

Alessandro Armuzzi, Rome, Italy

Digestive OncologyThomas Aparicio, Bobigny, France

Côme Lepage, Dijon, France

Tamara Matysiak-Budnik, Nantes, France

Pediatric GastroenterologySalvatore Cucchiara, Rome, Italy

General GastroenterologyElisabetta Buscarini, Crema, Italy

Cesare Hassan, Rome, Italy

HepatologyJérôme Dumortier, Lyon, France

Massimo Puoti, Milan, Italy

Sébastien Dharancy, Lille, France

Stefano Ginanni Corradini, Rome, Italy

Clinical Methodology Agostino Colli, Lecco, Italy

Digestive EndoscopyMassimo Raimondo, Jacksonville, USA

Pier Alberto Testoni, Milan, Italy

Alberto Larghi, Rome, Italy

Pancreatic DiseaseMassimo Falconi, Ancona, Italy

Gabriele Capurso, Rome, Italy

Basic ScienceGianfranco Alpini, Temple, USA

Romina Mancinelli, Rome, Italy

Editor EmeritusGabriele Bianchi-Porro, Milan, Italy

Editorial AssistantSerena Rotunno, Rome, Italy

Editor in ChiefMario Angelico, Rome, Italy

Managing EditorPaola Piccolo, Rome, Italy

Statistical ConsultantAlessandra Nardi, Rome, Italy

EDITORIAL BOARD

Domenico Alvaro, Rome, ItalyAngelo Andriulli, Foggia, ItalyPaolo Angeli, Padua, ItalyAdolfo Attili, Rome, ItalyGabrio Bassotti, Perugia, ItalyLaurent Beaugerie, Paris, FranceRobert Benamouzig, Bobigny, FranceAntonio Benedetti, Ancona, ItalyMarc Benninga, Amsterdam, NetherlandsMarina Berenguer, Valencia, SpainRoman Bogorad, Cambridge, USAJaime Bosch, Barcelona, SpainJean-Pierre Bronowicki, Vandoeuvre-Lès-Nancy, FranceWilliam Brugge, Boston, USANicola Caporaso, Naples, ItalyCarlo Catassi, Ancona, ItalyUmberto Cillo, Padua, ItalyDario Conte, Milan, ItalyGino Roberto Corazza, Pavia, ItalyEnrico Corazziari, Rome, ItalyAntonio Craxì, Palermo, ItalyRoberto De Franchis, Milan, ItalyGianfranco Delle Fave, Rome, Italy

Anthony Demetris, Pittsburgh, USASharon DeMorrow, Temple, USAPhilippe Ducrotte, Rouen, France Amal Dutta, Dallas, USALuca Elli, Milan, ItalyStefano Fagiuoli, Bergamo, ItalyLuigi Familiari, Messina, ItalyMassimo Fantini, Rome, ItalyPiero M. Fisichella, Boston, USAHeather Francis, Temple, USAMirella Fraquelli, Milan, ItalyDennis Freshwater, Birmingham, UKGiovanni B. Gaeta, Naples, ItalyAntonio Gasbarrini, Rome, ItalyEugenio Gaudio, Rome, ItalyShannon Glaser, Temple, USAPietro Invernizzi, Milan, ItalyRobert Jensen, Baltimore, USAMichel Kahaleh, Charlottesville, USADavid Laharie, Pessac, FranceRené Laugier, Marseille, FranceAstrid Lièvre, Saint-Cloud, FranceGiovanni Maconi, Milan, Italy

Riccardo Marmo, Salerno, ItalyMarco Marzioni, Ancona, ItalyDavid Mutimer, Birmingham, UKMattijs Numans, Leiden, NetherlandsJean-Marc Phelip, Saint Etienne, FranceAntonio Pinna, Bologna, ItalyFranco Radaelli, Como, ItalyAlessandro Repici, Milan, ItalyOliviero Riggio, Rome, ItalyMario Rizzetto, Turin, ItalyRenato Romagnoli, Turin, ItalyMassimo Rugge, Padua, ItalyTilman Sauerbruch, Bonn, GermanyJean-Christophe Saurin, Pierre-Benite, FranceVincenzo Savarino, Genoa, ItalyLaurent Siproudhis, Rennes, FranceEtienne Sokal, Brussels, BelgiumMario Strazzabosco, New Haven, USAGiacomo Carlo Sturniolo, Padua, ItalyGiuseppe Tisone, Rome, ItalyMichael Trauner, Vienna, AustriaVincenzo Villanacci, Brescia, ItalyFrank Zerbib, Bordeaux, France

Contents Vol . 47 Supplement 1 ( 15 February 2015 )

Index Medicus (MEDLINE), Current Contents/Clinical Practice,Science Citation Index and EMBASE/Excerpta Medica

Sociedad Iberoamericana de Información Cient ıfi ca (SIIC)

Associato alla Unione Stampa Periodica Italiana

Abstracts of the 48th A.I.S.F. – Italian Association for the Study of the Liver – Annual Meeting 2015: Oral communications e1

Abstracts of the 48th A.I.S.F. – Italian Association for the Study of the Liver – Annual Meeting 2015: Poster sessions – THURSDAY e19

Abstracts of the 48th A.I.S.F. – Italian Association for the Study of the Liver – Annual Meeting 2015: Poster sessions – FRIDAY e43

A.I.S.F. 2015: Abstracts evaluation procedure e67

Abstracts of the 48th A.I.S.F. – Italian Association for the Study of the Liver – Annual Meeting 2015

Rome, 19–20 February 2015

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Digestive and Liver Disease 47S (2015) e1–e18

Contents lists available at ScienceDirect

Digestive and Liver Disease

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bstracts of the 48th A.I.S.F. – Italian Association for the Study of theiver – Annual Meeting 2015: Oral communications

C-01

ELECTIVE LXR� INTESTINAL ACTIVATIONEDUCES HEPATIC INFLAMMATION ANDIBROSIS DURING THE DEVELOPMENT OFHRONIC LIVER INJURY

. Pierantonelli 1, C. Pinto 1, E. Mingarelli 1,. Saccomanno 1, L. Agostinelli 1, C. Rychlicki 1,. Trozzi 1, A. Benedetti 1, M. Marzioni 1,. Moschetta 2, G. Svegliati-Baroni 1

Dipartimento di Gastroenterologia, Universitàolitecnica delle Marche, Ancona, ItalyDipartimento di Medicina Interdisciplinare,niversità di Bari, Bari, Italy

Background: Hepatic fibrosis represents the wound-healingesponse of the liver to chronic injury characterized by increasednd altered deposition of newly generated extracellular matrix. Theathogenesis of liver fibrosis is not fully understood, leading to a

ack in effective therapies. Liver X receptors (LXR �/�) are impor-ant regulators of lipid metabolism. While in the liver LXRs regulateholesterol and fatty acid metabolism, intestinal LXR� activationas been implicated in reverse cholesterol transport (RCT) pathway

eading to high levels of the anti-inflammatory HDL.Aims: To evaluate the effect of the selective intestinal LXR� acti-

ation on the development of hepatic fibrosis associated to chroniciver injury.

Methods: Male FVB/N mice with intestinal constitutive LXR�ctivation (iVP16LXR�) and their control (iVP16) were treated withwice weekly i.p. injection of Carbon Tetrachloride (1 �l/g bodyeight) for 2 months.

Results: Immunohistochemistry for the macrophage marker4-80 indicated lowered infiltration in iVP16LXR� liver (p < 0.05).ene expression of the pro-inflammatory cytokines IL-6, TNF�,CP-1 and NF-Kb were reduced in iVP16LXR�-CCL4 mice (p < 0.05,< 0.05, p < 0.005 and p < 0.005 respectively). Collagen synthesisnd deposition was reduced in iVP16LXR� mice compared to iVP16s determined by type-I-collagen and TGF� mRNA expression (both< 0.005) and Sirius Red morphometry (p < 0.0005). RCT induced by

ntestinal LXR� activation was assessed by measuring plasmatic
DL concentration and found increased in iVP16LXR�-CCL4 mice
p < 0.005). Such activation is not associated to hepatic de novo

590-8658/$36.00

lipogenesis as shown by gene expression of SREBP1c and FAS anddecreased triglyceride content (p < 0.005).

Conclusions: Specific intestinal LXR� activation reduces liverinjury by increasing the level of the anti-inflammatory HDL choles-terol, thus leading to decreased hepatic fibrosis. Selective intestinalactivation of LXR� might be considered as a therapeutic approachto reduce liver fibrosis avoiding the occurrence of hepatic steatosisas a side effects associated to systemic LXR induction.

http://dx.doi.org/10.1016/j.dld.2015.01.007

OC-02

GUANFACINE (SPECIFIC ALPHA-2AADRENOCEPTOR AGONIST) RESTORESNATRIURESIS IN EXPERIMENTAL CIRRHOTICASCITES RESISTANT TO CLONIDINE ANDSTANDARD DIURETICS

G. Sansoè 1, M. Aragno 2, R. Mastrocola 2,M. Parola 2

1 Division of Gastroenterology, Gradenigo Hospital,Torino, Turin, Italy2 Department of Clinical and Biological Sciences,University of Torino, Turin, Italy

Introduction: In human cirrhosis, adrenergic hyperfunctioncauses proximal tubular fluid retention and contributes to refrac-tory ascites; clonidine, a sympatholytic drug, plus diureticsimproves natriuresis in otherwise refractory ascites.

Aim: To compare clonidine (aspecific �2-adrenoceptor agonist)and SSP-002021R (specific �2A-receptor agonist and prodrug ofguanfacine), both associated with diuretics, in cirrhotic refractoryascites.

Methods: Seven groups of 12 rats were studied: controls (G1);controls receiving furosemide and potassium canrenoate (G2); ratswith ascitic cirrhosis due to 14 CCl4 weeks (G3); cirrhotic ratstreated with furosemide and canrenoate over the 11th–14th CCl4weeks (G4); cirrhotic rats treated with furosemide, canrenoateand clonidine (0.5 mcg) (G5); cirrhotic rats treated with diureticsand low-dose clonidine (0.3 mcg) (G6); cirrhotic rats treated with
diuretics and SSP002021R (5 mg/kg b.w.) (G7). Three rats in eachgroup had their hormonal status and renal function assessed at theend of 11th, 12th, 13th, and 14th CCl4 weeks.

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Results: Cirrhotic rats in G3 and G4 gained weight over the1th–14th CCl4 weeks. In G4, brief increase in sodium excretionue to diuretics (12th week) preceded worsening of inulin clear-nce and natriuresis (diuretic resistance). The addition of low-doselonidine (G6) or guanfacine (G7) to diuretics increased, respec-ively, electrolytes excretion over the 11th–12th CCl4 weeks, orlomerular filtration rate (GFR) and electrolytes excretion over the3th–14th CCl4 weeks. Natriuretic responses in G6 and G7 wereccompanied by significantly reduced catecholamines serum levelss. G3 and G4 (all P < 0.03).

Conclusions: Clonidine briefly potentiates diuretics-dependentatriuresis. Conversely, guanfacine (�2A-receptor agonist) restoreslomerular filtration rate and natriuresis, and prevents refractoryscites in experimental advanced cirrhosis.

ttp://dx.doi.org/10.1016/j.dld.2015.01.008

C-03

HE PROTO-ONCOGENE TYROSINE-PROTEININASE MER AS A NOVEL MODULATOR OFIBROGENESIS IN NAFLD

. Di Maira 1, S. Petta 2, A. Cappon 1, E. Vivoli 1,

. Di Marco 2, F. Marra 1

Dipartimento di Medicina Sperimentale e Clinica,niversity of Florence, ItalyDipartimento Biomedico di Medicina Interna epecialistica, University of Palermo, Italy

Background and aims: MER (MERTK) is a receptor tyrosineinase of the TAM family often overexpressed or activated inarious malignancies. MERTK variants have been associated withbrosis severity at genome wide level in chronic hepatitis C, inarticular the variant rs4374383 G>A of MERTK gene has been asso-iated with reduced liver fibrosis progression. However, currentlyhere is no available experimental evidence on the involvement of

ERTK in the modulation of the biology of hepatic stellate cellsHSC) or in the fibrogenic process.

Methods: Primary human HSC were isolated from human liv-rs and cultured on plastic. C57BL6/J mice were treated with CCl4or 6 weeks. Balb/C mice were fed with a methionine and choline-eficient (MCD) diet for 8 weeks. Intrahepatic gene expression wasssayed by quantitative real time PCR.

Results: Using immunoprecipitation, we found the expressionf MERTK in human HSC. Stimulation of human HSC with theERTK ligand GAS6 resulted in a time-dependent activation of

RK1/2. Exposure to GAS6 also induced an enhancement of celligration of human HSC comparable to the effects of 10% FBS. In

oth models of fibrosis (chronic CCl4 and MCD diet) we observedsignificant increase of expression of MERTK compared to controlroup. Finally we studied the MERTK level in liver specimens ofatients with NASH with different degree of liver fibrosis. MERTKxpression was significantly higher in the group of patients withore severe fibrosis.Conclusions: In vitro and in vivo data indicate that MERTK could

lay an important role of in the biology of HSC and the fibrogenicrocess.


isease 47S (2015) e1–e18

OC-04

THE CYSTIC FIBROSIS CONDUCTANCE REGULAR(CFTR) CONTROLS C-SRC TYROSINE KINASESIGNALING AND REGULATES INNATE IMMUNITYAND EPITHELIAL POLARITY IN CHOLANGIOCYTES

R. Fiorotto 1, A. Villani 1, R. Scirpo 2, C. Spirli 1,M. Strazzabosco 1,2

1 Department of Internal Medicine, Liver Center andDigestive Diseases Section, Yale University, NewHaven, USA2 Department of Surgery and InterdisciplinaryMedicine, University of Milano-Bicocca, Milan, Italy

CFTR is expressed at the apical membrane of cholangiocyteswhere it regulates Cl− and HCO3

− secretion. CFTR also modulatesinnate immune responses in the biliary epithelium. In fact, TLR4-mediated responses to LPS are increased in cholangiocytes fromCftrtm1Unc (Cftr-KO) mice along with the activity of c-Src, a non-receptorial tyrosine kinase.

Aim: To understand how CFTR deficiency leads to up-regulationof c-Src activity in cholangiocytes.

Methods and results: Primary cholangiocytes were isolatedfrom Cftr-KO mice and their WT littermates. In Cftr-KO-cholangiocytes, c-Src inhibition by treatment with PP2 decreasedbasal and LPS-stimulated NF-�B activation and cytokines secretion.CFTR co-immunoprecipitated with proteins involved in the nega-tive regulation of c-Src (EBP-50, Csk and CBP); confocal imagingconfirmed their co-localization at the apical membrane in WT cells.Since c-Src is important for maintaining the integrity of epithelialcell–cell junctions, we investigated the distribution of ZO-1 (tightjunction), afadin (adherens junction) and subcortical F-actin fibers.In Cftr-KO cells ZO-1 and afadin lost their junctional restriction andalso appeared diffusely distributed in the cytoplasm; also the cor-tical actin ring failed to form properly, suggesting a polarity defect.Treatment with c-Src inhibitor PP2, rescued the polarity phenotype,as shown by ZO-1 and F-actin re-distribution. Inhibition of c-Srcin vivo significantly attenuated biliary damage and inflammationin a Cftr-KO mouse model.

Conclusions: Expression of CFTR is essential for the assembly ofan apical protein-complex located in lipid rafts that negatively reg-ulates c-Src. Lack of CFTR perturbs this complex. Consequently c-Srcself-activates promoting an increase in TLR4 responses, the desta-bilization of cell–cell junctions and an impairment in cell polarity.The protective effects of in vivo cSrc inhibition confirm the patho-

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expressing Omomyc; we engineered the cells also with luciferaseto follow cancer growth in vivo by bioluminescence imaging.


Abstracts / Digestive and L

C-05

ETFORMIN TARGETS A NEWZH2-PHOSPHOSTAT3-MIRNAS PATHWAY TONHIBIT LIPID DROPLETS ACCUMULATION ANDNTRACELLULAR INFLAMMATION

. Pediconi 1,2, S. Di Cocco 3, D. Salerno 3,. Belloni 2,3, S. Piconese 3, V. Barnaba 3,. Levrero 2,3

Department of Molecular Medicine, Sapienzaniversity, Rome, ItalySapienza Center for Life NanoScience, Rome, ItalyDepartment of Internal Medicine – DMISM,apienza University, Rome, Italy

Background and aim: Accumulation of triglyceride-containingipid droplets (LDs) within hepatocytes in NAFLD patients is aotentially reversible process, although sustained activation of

nflammatory signaling pathways leads to non-alcoholic steato-epatitis (NASH) that can evolve into cirrhosis and HCC. Here we

nvestigated the role of a new EZH2-phosphoSTAT3-miRNAs path-ay in LD accumulation and intracellular inflammation in an in vitro

ellular model of vescicularsteatosis.Methods: DMSO-differentiated human non-transformed Hep-

RG cells treated with oleic acid were used as a cellular model ofescicolarsteatosis.

Results: dHepaRG cells treated with oleic acid show: (a) anncreased lipid accumulation and intracellular reactive oxygenpecies (ROS) generation as compared to control cells; (b) dereg-lated lipid metabolism and liver-specific genes, including PDK4,LIN4, SLC2A1, ALB and ALDOB; (c) activation of an intracellular

nflammatory response, with the upregulation of IL6, IL8, OAS1,FKB and phosphoSTAT3. Oleate treatment also increased theRNA and protein levels of the EZH2 (Enhancer of Zeste Homolog

) histone methyl-transferase that catalyzes the repressive histonelysine 27 tri-methylation (H3K27me3). Treatment of oleate-

xposed HepaRG with S3I (STAT3 inhibitor) or MC1945 (new EZH2nhibitor) reduces triglycerides accumulation, represses metabolicenes over-expression and inhibits phosphoSTAT3 accumulation.e also found that several STAT3/IL6 responsive miRNAs, includingiR21 and mir24, are upregulated after lipid overload, paral-

eling STAT3 activation. Chromatin immuno-precipitation (ChIP)xperiments showed that the oleate-dependent transcriptionaleregulation of these miRNAs correlates with H3K27me3 levels,hosphoSTAT3 and EZH2 bound to their promoters. Moreover, met-ormin, an AMPK activator widely used as anti-diabetic drug andnown to improve lipid metabolism and to decrease steatosis innimal models, reduces phosphoSTAT3, inhibits miRNAs upregula-ion and reduces triglyceride accumulation.

Conclusions: We show that a new EZH2-phosphoSTAT3-iRNAs intracellular inflammation pathway amenable to thera-

eutic targeting is activated in well differentiated hepatocytes inesponse to lipid overload and is involved in vescicularsteatosis.


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OMOMYC AGAINST HCC: A MYC DOMINANTINTERFERING MINIPROTEIN TO COUNTERACTTUMOR GROWTH

B. Barbaro 1,2, C. Porcu 1, G. Toietta 3,R. Maggio 1,4, M. Savino 4, S. Nasi 4, C. Balsano 4

1 Francesco Balsano Foundation, Rome, Italy2 Department of Clinical and Applied Sciences andBiotechnology, L’Aquila University, L’Aquila, Italy3 Regina Elena Cancer Institute, Rome, Italy4 IBPM – CNR, Rome, Italy

Hepatocellular carcinoma (HCC) is the fifth most common tumorin the world for which the most promising strategy is targetedtherapy.

Abnormal Myc expression is linked to many solid tumors, butthis is usually not due to mutations in the Myc gene itself but resultsfrom upstream mutations. Then our strategy is to counteract itsaction avoiding its heterodimerization with Max, crucial for theDNA binding and consequent gene expression activation. To thisaim we used a dominant negative molecule, termed OMOMYC.

To note, also Hedgehog pathway, mostly Gli1 target gene, isreactivated during cancer, being normally switched-off in adults.Deregulation of Myc and Gli1 pathways is frequently observed inhuman hepatocarcinogenesis.

Aim: To determine whether HCC proliferation can be preventedby Omomyc and to ascertain whether this strategy can affect theHedgehog pathway.

Methods: We engineered HepG2 and Huh7.5.1 human hep-atoma cell lines to express Omomyc. We determined by microscopeexamination, MTS assay and cytofluorimetric analysis, morphol-ogy, proliferation and viability of cells with or without Omomycinduction. Real-time PCR and Western blot analysis were used fordetermining the expression of Myc, PCNA (Proliferating CellularNuclear Antigen), CCD1 (Cyclin D1), Hedgehog target gene Gli1.

Results: Omomyc is able to reduce HCC cell proliferation up to70% in 8 days. Analysis of mRNA expression of genes involved incell proliferation and corresponding protein levels confirmed theOmomyc effect in halting cell growth; moreover Omomyc was ableto decrease Gli1 expression.

Conclusion: Myc dominant negative miniprotein Omomyc isable to prevent HCC proliferation in vitro by specifically counter-acting the Hedgehog-mediated signaling.

We are now testing the outcome of this strategy in preventingtumor progression in a xenograft mouse model of HCC, using cells

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C-07

UT–LIVER AXIS DERANGEMENT DUE TO LACKF INFLAMMASOME ACTIVITY LEADS TOISCERAL OBESITY AND NASH DEVELOPMENT

. Agostinelli 1, C. Rychlicki 1, E. Mingarelli 1,. Saponaro 2, M. Gaggini 2, E. Buzzigoli 2,

. Pierantonelli 1, S. Saccomanno 1, C. Pinto 1,. Trozzi 1, A. Benedetti 1, S. De Minicis 1,. Marzioni 1, A. Gastaldelli 2, G. Svegliati-Baroni 1

Department of Gastroenterology, Polytechnicniversity of Marche, Ancona, ItalyInstitute of Clinical Physiology, National Researchouncil, Pisa, Italy

Introduction: NAFLD, the most common form of chronic liverisease, can lead to cirrhosis and hepatocellular carcinoma. Gutysbiosis and bacterial translocation induced by specific dietaryabits can elicit a proinflammatory and profibrogenic response.LRP3 inflammasome regulates intestinal homeostasis and medi-tes the release of proinflammatory cytokines in response toellular danger signals.

Aim: We want to investigate the role of NLRP3 inflammasomen a Western-lifestyle diet model of NAFLD.

Methods: Wild-type (WT) C57BL/6 and Nlrp3A350VneoR

Nlrp3−/−) mice were fed either a chow diet or a high-fat dietith fructose in drinking water (HFHC) for 12 weeks.

Results: Nlrp3−/− HFHC gained more weight (p < 0.01), showededuced energy expenditure and more fat mass (measured by dou-ly labeled water, all p < 0.05) with increased adipose tissue TNF�xpression (p < 0.05), and developed more hepatic steatosis (higherriglyceride content p < 0.01) compared to WT HFHC. Nlrp3−/−

FHC showed increased intestinal permeability, (i.e., lower cae-um zonulin-1 expression), that led to higher hepatic expression ofLR4 (p < 0.01) and TLR9 (p < 0.05). No differences were observedetween Nlrp3−/− HFHC and WT HFHC in hepatic expression ofREBP-1c effectors of de novo lipogenesis (ACC, FAS and SCD-1) thatere significantly increased in both groups. On the other hand,lrp3−/− HFHC showed higher expression of PPAR�2 (p < 0.01) andf its downstream effectors FABP-4 (p < 0.05) and CD36 (p < 0.01)hat regulate lipid uptake and storage. In addition Nlrp3−/− HFHChowed increased fatty acid mitochondrial-oxidation (i.e., higherPT1A expression) that, associated to the reduced expressionf NRF2 transcription-factor, led to increased superoxide pro-uction (measured by dihydroethidium-staining) (all p < 0.01).hese events were associated to increased macrophage infiltra-ion, collagen�1(I) and MCP1 gene expression (p < 0.05) in Nlrp3−/−

FHC mice only.Conclusions: In the Western-lifestyle diet, lack of NLRP3 inflam-

asome is associated with bacterial products translocation that
eads to metabolic alterations in both adipose tissue and liver, ando NASH development.

isease 47S (2015) e1–e18

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THE UK-PBC RISK SCORE: DERIVATION ANDVALIDATION OF A RISK SCORE TO PREDICT LIVEREVENTS IN THE UK-PBC RESEARCH COHORT

M. Carbone 1,2,3, S.J. Sharp 3, M.A. Heneghan 4,J.M. Neuberger 5, G.M. Hirschfield 6,A.K. Burroughs 7, D. Thorburn 7, A. Bathgate 8,M. Aldersley 9, C. Adgey 10, P. Trembling 11,K. Williamson 12, L. Jopson 13, R.T. Lim 5,N.J. Wareham 3, H.J. Cordell 14, G.J. Alexander 1,D.E. Jones 13, R.N. Sandford 2,G.F. Mells 1,2, UK-PBC Consortium

1 Division of Gastroenterology and Hepatology,Addenbrooke’s Hospital, Cambridge, United Kingdom2 Academic Department of Medical Genetics,University of Cambridge, Cambridge, UnitedKingdom3 MRC Epidemiology Unit, University of Cambridge,Cambridge, United Kingdom4 Institute of Liver Studies, King’s College Hospital,London, United Kingdom5 Liver Unit, Queen Elizabeth Hospital, UnitedKingdom6 NIHR Biomedical Research Unit and Centre for LiverResearch, University of Birmingham, Birmingham,United Kingdom7 The Sheila Sherlock Liver Centre, The Royal FreeHospital, London, United Kingdom8 Liver Unit, Royal Infirmary Hospital, Edinburgh,United Kingdom9 Liver Unit, St James’s Hospital, Leeds, UnitedKingdom10 Liver Unit, Royal Victoria Hospital, Belfast, UnitedKingdom11 Liver Unit, Barts and The London NHS Trust,London, United Kingdom12 Translational Gastroenterology Unit, JohnRadcliffe Hospital, Oxford, United Kingdom13 Institute of Cellular Medicine, NewcastleUniversity, Newcastle, United Kingdom14 Institute of Genetic Medicine, NewcastleUniversity, Newcastle, United Kingdom

Introduction: Outcomes in primary biliary cirrhosis (PBC) canbe predicted by biochemical response to ursodeoxycholic acid(UDCA). However, existing definitions do not take the stage of theliver disease into account and dichotomize UDCA response andlong-term risk whereas both are a continuum. We analyzed theUK-PBC Research Cohort to develop a risk score that includes mark-ers of disease stage, as well as post-treatment liver biochemistriesmodeled as continuous variables.

Methods: We constructed a PBC risk score for LT and liver-related death at 15 years in a derivation cohort (N = 2422) andevaluated it in a validation cohort (N = 1600). We used multivariablefractional polynomials (MFP) to model non-linear risk relationswith continuous variables, and 20 multiple imputations to replacemissing values. We fit a Cox proportional hazards model in eachimputed dataset, and used Rubin’s rules to combine the results.The resulting coefficients were used together with the baseline sur-vivor function to derive an equation for absolute risk at 15 years.
Net reclassification improvement (NRI) was calculated to comparethe predictive performance of this risk score compared with otherprognostic scores.

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Results: Albumin and platelet at baseline, ALP, bilirubin andransaminases after 12 months of UDCA were independently asso-iated with a liver event. The score incorporated these variables,ppropriately MFP-transformed. When applied to the validationohort, the score was highly predictive (c-statistic = 0.90). The NRIhowed that the risk score had greater ability to identify individ-als with and without events compared to other risk scores (NRIf PBC risk score vs. Paris1 = 90%; Paris2 = 64%; Barcelona = 38%;oronto = 71%).

Conclusions: Prognosis of UK patients with PBC can be accu-ately assessed with the PBC risk score by using readily availablebjective clinical measures. This may be used to identify high-riskatients for closer monitoring and second-line therapies, as wells low-risk patients who require infrequent monitoring and mightven be followed-up in primary care.


C-09

IVER DAMAGE CAN BE ASSOCIATED WITHEREGULATION OF THE DE NOVO LIPOGENESISATHWAY IN SUBJECTS WITH NON ALCOHOLICATTY LIVER DISEASE

. Rosso 1, C. Saponaro 2,3, L. Mezzabotta 1,

. Vanni 1, R. Gambino 1, F. Saba 1,

. Ibrahim Kamal Jouness 1, M. Gaggini 2,4,

. Buzzigoli 2, G.P. Caviglia 1, M.L. Abate 1,

. Salomone 5, A. Smedile 1, M. Rizzetto 1,. Cassader 1, A. Gastaldelli 2, E. Bugianesi 1

Department of Medical Sciences, University oforino, Turin, ItalyCardiometabolic Risk Unit, CNR-Institute of Clinicalhysiology, Pisa, ItalyDottorato Pegaso Regione Toscana in Biochimica eiologia Molecolare, University of Siena, Siena, ItalyDipartimento di Patologia Chirurgica, Molecolare ei Area Critica, University of Pisa, Pisa, ItalyU.O.C. of Gastroenterology, Azienda Sanitariarovinciale di Catania, Catania, Italy

Introduction: Adipose tissue insulin resistance (IR) and ele-ated adipose free fatty acid (FFA) flux are prominent features inAFLD. De novo lipogenesis (DNL) is up to 3-fold higher in sub-

ects with fatty liver and is not suppressed on fasting. When DNLs stimulated, the production of saturated FAs is increased and thexidation of FAs of any source is reduced. Both mechanisms canavor inflammation and IR.

Aim: We measured FFA flux/composition and used a surrogatendex of DNL (DNLi) to evaluate their relationship with histologicaleatures in a group of non-diabetic subjects with biopsy-provenAFLD.

Materials and methods: Hepatic and adipose tissue-IR indicesere derived from [2H5]glycerol and [2H2] glucose kinetics in a

roup of non-diabetic NAFLD patients in the basal state (n = 40)nd after a 4 h oral glucose load test (n = 20). Gas chromatographyass spectrometry was used to assess FFAs composition. DNLi was

erived as the ratio palmitic/linoleic acid.Results: Fasting plasma glucose/insulin, lipid profile, hep-

tic/adipose tissue IR indices and DNLi were similar in the tworoups. Fasting DNLi was associated with triglycerides (TG) andFAs levels and with adipose tissue IR (r = 0.597, r = 0.330 and
= 0.394, respectively). Among histological features, fasting DNLiignificantly correlated with steatosis (r = 0.364, P = 0.02) and NAScore (r = 0.306, P = 0.05). After the glucose load, TG levels initially
isease 47S (2015) e1–e18 e5

increased despite elevated insulin levels, suggesting a significantcontribution of DNL. Accordingly, DNLi consensually increased withTG levels (r = 0.749, P < 0.005) and was significantly related to thedegree of fibrosis (rs = 0.514, P = 0.02).

Conclusions: Oral glucose load is associated with changes inDNL and hepatic triglyceride synthesis that can favor liver fibrosisin patients with NAFLD.

Funding: Funded by FP7/2007-2013 under grant agreementn◦ HEALTH-F2-2009-241762 for the project FLIP and by PRIN2009ARYX4T.


OC-10

THE PRESENCE OF WHITE MATTER LESIONS ISNOT ASSOCIATED WITH NON-ALCOHOLIC FATTYLIVER DISEASE BUT WITH ITS HISTOLOGICALSEVERITY

S. Petta 1, A. Tutolomondo 2, C. Gagliardo 4,R. Zafonte 5, G. Brancatelli 4, D. Cabibi 3,C. Cammà 1, V. Di Marco 1, L. Galvano 5, A. Licata 2,F. Magliozzo 5, G. Merlino 5, M. Midiri 4, A. Pinto 2,A. Craxì 1

1 Sezione di Gastroenterologia, DiBiMIS, University ofPalermo, Italy2 Dipartimento Biomedico di Medicina Interna eSpecialistica (Di.Bi.M.I.S), Università di Palermo,Palermo, Italy3 Cattedra di Anatomia Patologica, University ofPalermo, Palermo, Italy4 Department of Radiology, University of Palermo,Palermo, Italy5 Medicina Generale Palermo, Palermo, Italy

Background: Nonalcoholic fatty liver disease (NAFLD) has beenassociated with increased cardiovascular risk, including coronaryartery disease and cerebrovascular events. No studies howeverassessed the potential relationship between NAFLD and subclini-cal cerebrovascular alterations. We tested the correlation betweenNAFLD and its histological severity with vascular white matterlesions (WML) in patients with biopsy-proven NAFLD and in nonsteatosic controls.

Methods: The anthropometric, biochemical and metabolic fea-tures were recorded in 77 consecutive biopsy-proven NAFLD(Kleiner score), and in 35 controls with normal ALT, without chronicliver diseases, and without ultrasonographic evidence of steatosis.All patients underwent magnetic resonance assessment of WML.WML were classified according to the Fazekas score in absent (0/III),or present (mild I/III; moderate II/III, and severe I/III). For purpose ofanalyses all controls, as plausible, were considered without NASHand without F2-F4 liver fibrosis.

Results: WML were found in 26% of the entire cohort (29/112),even if of a moderate–severe grade in 5 patients only. The preva-lence of WML was similar in NAFLD compared to no NAFLD (27% vs23%; p = 0.62). Age ≥50 yrs, female gender, type 2 diabetes, arterialhypertension, presence of NASH (35% vs 18%, p = 0.05) and presenceof F2-F4 fibrosis (43% vs 17%, p = 0.003) were associated with WMLpresence (p = <0.01). At multivariate analysis age >50 yrs (OR 3.4495% CI 1.01–11.6. p 0.04), female gender (OR 3.71. 95% CI 1.28–10.7.p 0.01), and F2-F4 fibrosis (OR 3.39. 95% CI 1.17–9.84. p 0.02) weremaintained as factors independently associated with WML. When
considering NAFLD patients only, we confirmed F2-F4 fibrosis asthe only independent predictor of WML (OR 4.24. 95% CI 1.14–15.7.p 0.03).

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Conclusion: The presence of WML is not associated with NAFLDut with its histological severity. Clinical implications of this issueeed to be assessed by longitudinal studies.


C-11

ONG-TERM USE OF HUMAN ALBUMIN FOR THEREATMENT OF ASCITES IN PATIENTS WITHEPATIC CIRRHOSIS: THE INTERIM ANALYSIS OFHE ANSWER STUDY

. Bernardi 1, O. Riggio 2, P. Angeli 3,. Alessandria 4, S. Neri 5, F.G. Foschi 6,. Levantesi 7, S. Boccia 8, A. Airoldi 9, S. Fagiuoli 10,. Svegliati-Baroni 11, G. Laffi 12, R. Cozzolongo 13,. Butera 14, V. Sangiovanni 15, P. Toniutto 16,.A. Zocco 17, R. De Marco 18, F. Morisco 19,

. De Leonardis 20, I. Cacciola 21, G. Elia 22,. Federico 23, S. Massironi 24, R. Guarisco 25,. Marin 26, S. Piano 3, C. Elia 4, S. Nardelli 2,. Maiorca 5, E. Neri 6, A. Mastroianni 7,. Tufoni 1, L. Simone 8, L. Cesarini 9, G. Magini 10,. Marzioni 11, R.G. Romanelli 12,. Zappimbulso 13, F. Macaluso 14, G. Parrella 15,

. Pugliese 16, A. Tortora 17, M. Cavallin 3,. Andrealli 4, C. Pasquale 2, F. Fidone 5, A. Lanzi 6,. Alberti 9, F. Salerno 27, A. Roncadori 28,. Zaccherini 1,. Caraceni 1, the ANSWER Study Group

Università di Bologna, ItalyUniversità di Roma “La Sapienza”, ItalyUniversità di Padova, ItalyA.O. Città della Salute e della Scienza di Torino, ItalyUniversità di Catania, ItalyA.U.S.L. Ravenna – Osp. di Faenza, ItalyA.U.S.L. Bologna – Osp. di Bentivoglio, ItalyA.O.U. di Ferrara, ItalyA.O. Niguarda Ca’ Granda di Milano, Italy

0 A.O. Papa Giovanni XXIII di Bergamo, Italy1 Università di Ancona, Italy2 Università di Firenze, Italy3 IRCSS “De Bellis” di Castellana Grotte, Italy4 Università di Palermo, Italy5 A.O.R.N. dei Colli – Osp. Cotugno di Napoli, Italy6 Università di Udine, Italy7 Università Cattolica di Roma, Italy8 A.O. di Cosenza, Italy9 Università di Napoli “Federico II”, Italy0 Università di Roma “Tor Vergata”, Italy1 Università di Messina, Italy2 A.O.U. di Parma, Italy3 Seconda Università di Napoli, Italy4 Università di Milano – Osp. Maggiore Policlinico,taly5 A.S.L. Roma H – Osp. “San Giuseppe” di Marino,taly6 A.U.L.S.S. Mirano – Osp. di Dolo, Italy7 Università di Milano – Policlinico San Donato, Italy8 C.I.N.E.C.A. – Bologna, Italy

Introduction: Despite long-term human albumin (HA) adminis-ration for treating cirrhosis with ascites is commonly used in Italy,
he scientific evidence of its efficacy is still lacking.
Aim: To assess the efficacy of long-term HA administration inatients with cirrhosis and ascites.

isease 47S (2015) e1–e18

Methods: In this multicentre, prospective, randomized clinicaltrial, 420 patients with cirrhosis and uncomplicated ascites treatedwith anti-mineralocorticoids (≥200 mg/day) and furosemide(≥25 mg/day) are planned to be randomized 1:1 to either standardmedical treatment (SMT) or SMT + HA (40 g twice weekly for 2weeks, and then 40 g weekly). Death, liver transplantation, TIPS,refractory ascites requiring ≥3 paracentesis/month or 18 monthsfollow-up terminate the study. The primary end-point is mortality.Among the secondary end-points, those related to the manage-ment of ascites and the incidence of complications of cirrhosis wereassessed in this analysis.

Results: 386 (SMT: 188; SMT + HA: 198) patients were included.Their respective median follow-up duration was 183 (129–233)and 301 (238–355) days (p = 0.021). Baseline demographics andclinical and laboratory parameters were well balanced betweenthe two arms. Kaplan–Meier intention-to-treat analysis showedthat mortality was significantly reduced in patients receivingHA (at 18 months: SMT + HA: 22%, SMT: 34%; p = 0.045). Statisti-cally significant benefits were found in the SMT + HA arm in theincidence rate of paracentesis (−55%, p < 0.001), incidence of refrac-tory ascites (−42%, p < 0.001), and need of ≥3 paracentesis/month(−62%; p < 0.001). HA arm also presented an advantage in theincidence rates of SBP (−57%; p = 0.004), hepatic encephalopathygrade III–IV (−37%; p = 0.005) and renal impairment (serum cre-atinine > 1.5 mg/dl) (−28%; p = 0.011). HA administration did notincrease the risk of variceal bleeding, and the incidence of severeadverse effects was similar in the two arms.

Conclusions: Long-term HA administration prolongs survival inpatients with cirrhosis and ascites, improves greatly the manage-ment of ascites and reduces the incidence of severe complicationsof the disease.


OC-12

VALIDATION OF A NEW CHILD-TURCOTTE-PUGHCLASS 0 FOR PATIENTS WITH HEPATOCELLULARCARCINOMA IN A EUROPEAN COHORT

A. Gianstefani, A. Pecorelli, I. Pettinari,L. Venerandi, F. Piscaglia,L. Bolondi, the ITA.LI.CA. Study Group

Department of Medical and Surgical Sciences,S.Orsola-Malpighi Hospital, Alma Mater Studiorum –University of Bologna, Bologna, Italy

Background: Hepatocellular carcinoma (HCC) often arises inthe context of very well-preserved liver function, expressed bya Child-Turcotte-Pugh (CTP) class A, which comprises only twoscores, 5 and 6. Recent published data identified a new CTPscore, named 0, defined by fulfilling all the subsequent criteria:albumin ≥ 4 g/dL, bilirubin ≤ 0.8 mg/dL, prothrombin time prolon-gation < 0 s, no ascites, no encephalopathy. In an Asiatic population,this subgroup of HCC patients with very well-preserved liver func-tion had a better outcome compared to the CTP class A patients.

Aim: To validate the prognostic role of the new CTP 0 in a Euro-pean cohort of HCC patients.

Methods: A total of 4248 patients with a first diagnosis ofHCC enrolled in the ITA.LI.CA. database between 1986 and 2012were retrospectively analyzed. Patients were divided in four groupsaccording to the CTP class: 0 (232 pts), A (2514 pts), B (1221 pts)and C (281 pts). Patient survival probability was estimated using

Results: The 8.4% of CTP A patients were reclassified as CTP0. Median overall survivals statistically differed among groups

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months; 95% CI): CPT 0 62 (52.9–71.1), A 44 (41.6–46.4), B 2219.7–24.3), C 9 (6.6–11.3), p < 0.0001. Comparisons between sur-ivals of CTP 0 vs A, B and C were also statistically differentp < 0.0001 in all associations). The prognosis of patients in thentermediate BCLC stage also differed according to the liver function0 vs A vs B, p < 0.0001).

Conclusions: The newly proposed CTP class 0 identifies a differ-nt subgroup of patients with a better prognosis, also when appliedn a European cohort, where HCV aetiology is predominant.

This new approach impacts not only on outcome prediction butlso, potentially, on treatment allocation, better stratifying, in par-icular, the heterogeneous intermediate BCLC-B stage.


C-13

ATHOGENESIS AND CLINICAL IMPACT OFELATIVE ADRENAL INSUFFICIENCY INOSPITALIZED PATIENTS WITH ACUTEECOMPENSATION OF CIRRHOSIS

. Piano 1,2, E. Favaretto 2, S. Fasolato 1,2,. Scaroni 2, E. Gola 2, A. Brocca 2, A. Sticca 2,. Morando 2, M. Cavallin 2, A. Romano 2,. Gringeri 3, A. Gatta 2, U. Cillo 3, M. Plebani 2,. Angeli 1,2

Unit of Hepatic Emergencies and Liverransplantation, University of Padua, Padua, ItalyDepartment of Medicine, University of Padua,adua, ItalyUnit of Hepatobiliary Surgery and Liverransplantation, University of Padua, Padua, Italy

Background and aims: Relative adrenal insufficiency (RAI) haseen described in patients with cirrhosis and has been recentlyssociated with the development of hepatorenal syndrome, sepsis,nd poor survival. Its pathogenesis has not yet been clearly defined.he aim of our study was to explore factors associated with RAI ando investigate clinical impact of RAI in these patients.

Methods: 94 patients admitted to the hospital for an acuteecompensation of cirrhosis were consecutively enrolled in thetudy and followed up for 90 days. Adrenal function was assessedith short synacthen test (SST). RAI was diagnosed when the

ncrease in serum total cortisol after SST was <9 �g/dL in patientsith basal serum total cortisol < 35 �g/dL. Bacterial infections,arkers of inflammations, ACTH, substrates for steroidogenesis

nd apolipoprotein A1 (ApoA1), which is required for cholesteryl-ster accumulation in steroidogenic cells, were explored as possibleathogenetic factors involved in RAI.

Results: RAI was diagnosed in 42.6% of patients. Those withAI were younger (57.0 vs 61.5 years; p = 0.047), had higherELD-Na score (22 vs 18.4; p = 0.01), higher C-reactive protein

15.0 vs 11.5 mg/L; p = 0.047), and lower total cholesterol (1.5s 2.1 mmol/L; p = 0.028), HDL (0.4 vs 0.6 mmol/L; p = 0.034) andpolipoprotein A1 (0.6 vs 0.8; p = 0.006) than those without RAI.NF-alpha, IL6, IL-10, IL1 beta and ACTH were not significantlyifferent between the two groups. ApoA1 was the only inde-endent predictor of RAI (OR = 0.12; p = 0.011). In our series RAIas associated with a higher risk to develop bacterial infections

65.1 vs 42.4%; p = 0.039), hyponatremia (46.0 vs 14.3%; p = 0.002),nd with poorer 90-day transplant free survival (70.7 vs 90.3%;= 0.013). In multivariate analysis MELD-Na (HR = 1.096; p = 0.030),
ge (HR = 1.059; p = 0.043) and RAI (HR = 3.277; p = 0.041) wereound to be independent predictors of mortality.
isease 47S (2015) e1–e18 e7

Conclusions: RAI is frequent in patients hospitalized for anacute decompensation of cirrhosis. Low level of ApoA1 seems tohave a pivotal role in its pathogenesis. RAI is associated with a highrisk to develop bacterial infections, hyponatremia and with a lowprobability of survival in these patients.


OC-14

COST-EFFECTIVENESS OF PRE-TRANSPLANTSOFOSBUVIR TO PREVENT RECURRENCE OF HCVINFECTION AFTER LIVER TRANSPLANTATION

A. Vitale 1, G. Spolverato 1, P. Burra 1,T.M. De Feo 2, U. Cillo 1, S. Fagiuoli 3, On behalf ofthe Liver Transplantation NITp Working Group

1 University Hospital of Padua, Padua, Italy2 North Italy Transplant Program, Fond. IRCCS Ca’Granda OMP, Milan, Italy3 Gastroenterology and Transplant Hepatology,Ospedale Papa Giovanni XXIII, Bergamo, Italy

Background and aims: The strong efficacy of pre-transplantSofosbuvir (SOF) plus Ribavirin in preventing recurrence of hep-atitis C virus (HCV) infection after liver transplantation (LT) hasbeen reported. The aim of this study was to evaluate the cost-effectiveness of this strategy in the North Italy Transplant program(NITp) area.

Methods: We first evaluated the impact of HCV infection onpost-LT survival in 2376 consecutive adult patients (MELD ≤ 25,period 2004–2009, NITp area) and prevalence-costs of conven-tional standard of care (SOC) antiviral therapy (Peginterferon plusribavirin) after LT. A Markov model was developed to comparedtwo strategies: SOF + Ribavirin (RBV) as pre-LT anti-HCV treat-ment (strategy A), versus on-demand post-LT SOC antiviral therapy(strategy B). The endpoint was incremental cost-effectiveness ratio(ICER) as costs ($)/quality-adjusted life year (QALY).

Results: Among the 1794 patients undergoing LT and meet-ing the inclusion criteria, 860 (48%) were HCV+ and 50% of themreceived SOC therapy post-LT (drugs and adverse effects manage-ment mean costs = 16,440$ for patient). HCV etiology had a strongimpact on post-LT survival (Hazard Ratio = 1.59, 95% CI = 1.22–2.09,p = 0.0007). After Monte Carlo simulation, pre-LT SOF therapyshowed a median survival benefit of 2.0 QALYs, and an ICER of33,600$/QALY. Costs of SOF therapy, sustained viral response rate12 weeks after LT, and recipient age were the main ICER predictorsat multivariate analysis.

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C-15

NOS POLYMORPHISMS IN RELATION TOUTCOME IN ADVANCED HCC PATIENTSECEIVING SORAFENIB

. Casadei Gardini 1,∗, G. Marisi 1, E. Scarpi 1,. Scartozzi 2, F.G. Foschi 3, L. Faloppi 3, E. Tenti 1,

. Tamberi 3, E. Tamburini 3, G.L. Frassineti 1,

. Cascinu 2, P. Ulivi 1

IRST-IRCCS, Meldola, ItalyDepartment of Medical Oncology, A.O. Ospedaliiuniti Università Politecnica delle Marche, Ancona,

talyOspedale per gli Infermi, Faenza, Italy; AUSLavenna, Faenza, Italy

Background and aims: Cancer cells adapt to hypoxic microen-ironment through the activation of many molecules, includingndothelial nitric oxide synthase (eNOS). Sorafenib, by blocking theascular endothelial growth factor receptors (VEGFRs), induces annhibition of eNOS activity with a consequent decrease of the pro-uction of nitric oxide (NO). NO is associated with an increase ofumor angiogenesis, tumor invasion and metastasis formation. Inur study we analysed the role of eNOS polymorphisms in rela-ion to clinical outcome in patients with hepatocellular carcinomareated with sorafenib.

Methods: From a database of 257 patients diagnosed with hepa-ocellular carcinoma from 2004 to 2014, we selected 54 patientsho received sorafenib. Peripheral blood samples or FFPE tumor

issues were available for DNA extraction and genotyping analy-is. Three eNOS polymorphisms (eNOS +894 G/T, eNOS VNTR 27 bpa/b, eNOS −786 C/T) were analyzed by direct sequencing or Realime PCR method. We analyzed 21 patients for the VNTR 4a/b poly-orphism and 32 patients for −786 C/T polymorphism. All the

andidate genotypes were evaluated to identify a potential corre-ation with overall survival (OS) (log-rank test).

Results: With regard to eNOS VNTR it was observed that patientsarrying the b allele (5 repetitions of 27 bp) both in homozygosity4bb) and in heterozygosity (4ab) were associated with a betterS. The variants 4aa (4 repeats of 27 bp in homozygosity), 4abnd 4bb, were associated with a median OS of 5.7, 13.9 and 23.6onths, respectively (p = 0.016). For eNOS-786 the presence of theallele both in homozygosity (TT) and in heterozygosity (TC) was

ssociated with a statistically significant longer OS with respecto patients with CC genotype (15.6 versus 13.9 months, respec-ively, p = 0.031). No correlations were observed in relation to PFSp = 0.494).

Conclusions: eNOS VNTR and eNOS −786 could represent
rognostic markers in patients with advanced hepatocellular car-inoma treated with sorafenib.

isease 47S (2015) e1–e18

OC-16

THE AAA+ ATPASE RUVBL1 COORDINATES LIVERMETABOLISM AND HEPATOCELLULARCARCINOMA PROGRESSION

T. Mello 1,2, M. Materozzi 1, F. Zanieri 1,O. Bereshchenko 3, E. Ceni 1, M. Tarocchi 1,2,G. Marroncini 1, I. Simeone 1, S. Polvani 1,S. Tempesti 1, C. Nerlov 4, S. Milani 1,2, A. Galli 1,2

1 Department of Biomedical, Experimental andClinical Sciences “Mario Serio”, University ofFlorence, Florence, Italy2 Careggi University Hospital, Florence, Italy3 Department of Medicine, Section of Pharmacology,University of Perugia, Perugia, Italy4 Institute for Stem Cell Research and MRC Centre forRegenerative Medicine, University of Edinburgh,Edinburgh, UK

The AAA+ ATPase Ruvbl1 is overexpressed in several humancancers, including hepatocellular carcinoma (HCC), in which highRuvbl1 expression correlates with a poor prognosis. A growing bodyof data from in vitro models supports the concept that dysregulationof Ruvbl1 expression occurs in cancer to promote its growth andprogression, making this protein an attractive target for anti-cancertherapies. However, whether Ruvbl1 participate in the oncogenictransformation and cancer progression in vivo remains speculative.To challenge this question we realized a hepatocyte-conditionalRuvbl1 hemizygous mouse and evaluated the tumor onset and pro-gression after DEN injection.

Ruvbl1 hemizygous mice were obtained by crossing Ruvbl1-floxed with Albumin-Credeleter mice. The male offspring weresubjected to a single i.p. injection of DEN (5 mg/kg) to induce livercancer, and were monitored up to one year after cancer induction.

Conditional liver-hemizygous mice showed no obvious pheno-type with respect to liver size, viability and liver function (AST-ALT),however they had significant alterations of glucose, triglyceridesand cholesterol serum levels, and an increased body weight.

Despite a significant delay in the onset of liver cancer, Ruvbl1+/−mice eventually developed significantly larger tumors than con-trol mice. We found that Ruvbl1+/− mice had reduced hepatocyteturnover and impaired mTOR signaling than wild-type mice, whichis likely causing both the metabolic alterations and the delayedonset of HCC.

In conclusion, this is the first report highlighting a role ofRuvbl1 as a major regulator of hepatic metabolism. Contrary to ourexpectations, we found that although in the hepatocyte-conditionalRuvbl1+/− mice the onset of HCC is delayed, its progressionis accelerated. The underlying molecular mechanisms are underinvestigation, nevertheless this report highlights the potential risksof prolonged Ruvbl1 inhibition in a intact mammalian organism.

Acknowledgement: This research is funded by the Italian Min-istry of Health through grant GR-2009-1600315.


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high risk for EKD should be considered for alternative treatmentstrategies, including combined liver-kidney transplantation.



C-17

HE EMPIRICAL ANTIBIOTIC TREATMENT OFOSOCOMIAL SPONTANEOUS BACTERIALERITONITIS IN PATIENTS WITHECOMPENSATED CIRRHOSIS: RESULTS OF AANDOMIZED CONTROLLED CLINICAL TRIAL

. Piano 1,2, F. Salinas 3, S. Fasolato 1,2,

. Morando 2, M. Cavallin 2, A. Romano 2, S. Rosi 2,. Stanco 2, A. Sticca 2, M. Senzolo 4, P. Burra 4,. Zanus 5, U. Cillo 5, A. Gatta 2, P. Angeli 1,2

Unit of Hepatic Emergencies and Liverransplantation, University of Padua, Padua, ItalyDepartment of Medicine, University of Padua,adua, ItalyDivision of Medicine, Private Hospital “GiovanniXIII” of Monastier, Treviso, ItalyMultivisceral Transplant Unit, University of Padua,adua, ItalyUnit of Hepatobiliary Surgery and Liverransplantation, University of Padua, Padua, Italy

Background and aims: Spontaneous bacterial peritonitis (SBP)s a common and life-threatening complication of cirrhosis. Thirdeneration cephalosporins (TGCs) are the first line empirical treat-ent of SBP. In recent years, an increasing rate of SBP due to TGCs

esistant bacteria has been observed, in particular in nosocomialpisodes. Currently a broader spectrum antibiotic regimen such asarbapenems and glycopeptides/lipopeptides has never been com-ared to TGCs in the treatment of nosocomial SBP. The aim of ourtudy was to compare the efficacy of meropenem plus daptomycinersus ceftazidime in the treatment of nosocomial SBP.

Methods: Patients with cirrhosis, ascites and nosocomial SBPere randomized to receive meropenem (1 g/8 h) plus daptomycin

6 mg/kg/day) or ceftazidime (2 g/8 h) plus albumin in both groups.diagnostic paracentesis was performed after 48 h of antibiotic

reatment. A reduction in ascitic fluid neutrophil count <25% of there-treatment value was considered a treatment failure and antibi-tic therapy was changed accordingly. The primary outcome washe efficacy of the treatment defined by the resolution of SBP afterdays of treatment (NCT01455246).

Results: 32 patients were randomized and 31 were analyzed.he combination of meropenem plus daptomycin was significantlyore effective than ceftazidime in the treatment of nosocomial

BP (86.7 vs 25%; p < 0.001). TGCs resistant bacteria were isolatedn 81.3% of positive cultures. In multivariate analysis, ineffec-ive response to first line treatment (hazard ratio [HR] = 20.6;= 0.01), development of acute kidney injury during hospitalization

HR = 23.2; p = 0.01) and baseline mean arterial pressure (HR = 0.92;= 0.01) were found to be independent predictors of 90-day trans-lant free survival. The incidence of adverse events was similaretween the two groups.

Conclusions: The combination of meropenem plus daptomycins more effective than ceftazidime in the treatment of hospital
cquired SBP. The efficacy of the first line treatment is associatedith improved 90-day survival in these patients.

isease 47S (2015) e1–e18 e9

OC-18

RISK FACTOR FOR EARLY KIDNEY DISEASEAFTER LIVER TRANSPLANTATION AND ITSIMPACT ON GRAFT LOSS: INSIGHTS FROM THELIVER MATCH COHORT STUDY

S. Ginanni Corradini 1, A.P. Mitterhofer 2,A. Nardi 3, T. Marianelli 4, L. Parlati 1, F. Tinti 2,M. Angelico 4, the Liver Match Study Group

1 Gastroenterology, Sapienza Università di Roma,Italy2 Nephrology, Sapienza Università di Roma, Italy3 Biostatistics, Università di Tor Vergata, Roma, Italy4 Hepatology and Transplant, Università di TorVergata, Roma, Italy

Introduction and aim: Early Kidney Dysfunction (EKD) is fre-quent after liver transplantation (LT). We investigated the riskfactors for EKD and its effect on graft loss.

Methods: We evaluated 1302 patients undergoing LT in Italyfrom June 1, 2007 to May 31, 2009 belonging to the LiverMatch cohort. EKD was staged based on the lowest estimatedglomerular filtration rate (eGFR, MDRD-4 formula) at 3 and 6months after LT, as follows: eGFR > 60 ml/min (EKD stage 1-2);45 ≤ eGFR < 60 ml/min (EKD stage 3a); 30 ≤ eGFR < 45 ml/min (EKDstage 3b); and eGFR < 30 ml/min (EKD stage 4–5).

Results: 812 patients had EKD stage 1–2 (62.4%), 404 EKD stage3a/b (31.0%) and 86 EKD stage 4–5 (6.6%). Multinomial logistic anal-ysis identified four recipient risk factors for EKD: old age, femalegender, and low eGFR and sodium levels at LT (table).

EKD stage 3a/b vs stage 1–2 EKD stage 4–5 vs stage 1–2

Odds ratio 95% C.I. p-Value Odds ratio 95% C.I. p-Value

Recipients age(×10 years)

1.63 1.40–1.89 <0.0001 1.70 1.28–2.27 0.0003

Recipient gender(F vs M)

2.29 1.68–3.12 <0.0001 2.00 1.58–3.46 0.0129

eGFR at LT 30–60vs ≥60 ml/min

2.55 1.79–3.64 <0.0001 2.72 1.50–4.94 0.0010

eGFR at LT < 30 vs≥60 ml/min

2.82 1.35–5.89 0.0059 9.74 4.08–23.28 <0.0001

Recipient sodiumat LT (per5 meq/L)

0.86 0.75–0.98 0.0252 0.73 0.58–0.92 0.0082

Multivariable Cox regression identified EKD stage as a signif-icant risk factor for 5-year graft loss. Assuming EKD stage 1–2as reference level, the hazard ratio of graft loss for EKD stage3a, 3b, and 4–5 were 0.89 (p = 0.4729), 1.68 (p = 0.0059) and 3.38(p < 0.0001), respectively.

Conclusions: The probability of EKD can be estimated beforeLT and EKD stage > 2b entails a high risk of graft loss. Patients at

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C-19

REVENTION OF BLEEDING FOLLOWINGNVASIVE PROCEDURES IN CIRRHOTICATIENTS: A SINGLE CENTER EXPERIENCE

. Tosetti 1, F. Invernizzi 1, V. La Mura 1,

. Aghemo 1, M. Primignani 1, A. Sangiovanni 1,.F. Donato 1, A. Nicolini 2, M. Iavarone 1,. Colombo 1

Division of Gastroenterology and Hepatology,ondazione IRCCS Cà Granda Ospedale Maggioreoliclinico, University of Milan, Milan, ItalyDivision of Radiology, Fondazione IRCCS Cà Grandaspedale Maggiore Policlinico, University of Milan,ilan, Italy

Background: Risk of bleeding in cirrhosis has predominantlyeen associated with coagulopathy and thrombocytopenia due to

mpaired liver function and splenomegaly. The evaluation of thectual bleeding risk in cirrhotic patients undergoing invasive proce-ures is a critical point to optimize management in terms of plateletPlt) or plasma prophylactic transfusions.

Methods: During 2013, 480 cirrhotic patients underwentiagnostic or therapeutic invasive procedures at our Liver UnitFondazione IRCCS Cà Granda, Ospedale Maggiore Policlinico,

ilan, Italy). Plt transfusion was performed when Plt count lesshan 50,000/mmc while fresh frozen plasma was infused whennternational normalized ratio (INR) > 1.5, except those undergoingaracentesis. Low-molecular-weight-heparin was discontinued4 h before any procedure while antiaggregant therapy was inter-upted five days before any procedure except oesophageal varicesand ligation and paracentesis. Major hemorrhagic events werehose requiring hospitalisation or blood transfusion; minor eventsas a haemoglobin decline > 1.5 g/dl post procedure without clini-

al relevance.Results: 174 Transarterial Chemo-Embolization (TACE), 16

adio-Frequency Termal Alblation (RFTA), 214 paracentesis, 59esophageal varices banding, 6 trans-jugular liver biopsy (LB) and1 Percutaneous Ethanol Injection (PEI) were performed. Over-ll, 61 procedures met the criteria for plasma or Plt infusion andajor bleeding complications occurred in 3 patients (0,6%). In 2

atients, anemia-related paracentesis was treated by blood trans-usions whereas one patient following variceal band ligation hado be hospitalized for severe anemia. Major complications rate was.6% in Plt/plasma infusion exposed patients versus 0.47% in unex-osed (p = 0.28). Minor events with a > 1.5 g/dl haemoglobin declineccurred in 17 patients (15 paracentesis and 2 band ligations), rateas 3.2% in Plt/plasma infusion exposed patients versus 3.5% innexposed (p = 0.9).

Conclusion: Pre-treatment platelet transfusion in cirrhoticatients with Plt count < 50,000/mmc and plasma infusion for

NR > 1.5, was associated with a negligible risk of bleeding andppeared as a safe, cost/effective strategy.


isease 47S (2015) e1–e18

OC-20

ANTI CAPSID DRUGS HAP12 AND AT130 TARGETHBV CORE PROTEIN NUCLEAR FUNCTIONS

L. Belloni 1, G.A. Palumbo 2, L. Li 3, S.R. Chirapu 4,L. Calvo 1,2, L. Lupacchini 1, M.G. Finn 4,U. Lopatin 5, A. Zlotnick 3,5, M. Levrero 2

1 Centre for Life Nanoscience, IIT-Sapienza, Rome,Italy2 Department of Internal Medicine – DMISM,Sapienza University of Rome, Italy3 Department of Molecular & Cellular Biochemistry,Indiana University, Bloomington, USA4 Department of Chemistry, Georgia Institute ofTechnology, Atlanta, USA5 Assembly Bioscience, Bloomington, USA

Introduction and aim: HBV core protein (Cp) represents anattractive new therapeutic target for HBV chronic infection. Cphas been shown to bind the nuclear cccDNA mini-chromosome aswell as a number of cellular genes promoters. Several compoundsthat target Cp and HBV capsids assembly, including the hetero-aryl-dihydropyrimidines (HAPs) and the phenyl-propenamidederivatives AT61 and AT130, have been shown to inhibit HBV repli-cation in vitro and in vivo. HAPs and AT130 enhance the rate of Cpassembly and stabilize preferentially non-capsid polymers of Cp.Here we investigated the ability of the Core protein Assembly Mod-ulators (CaMPs) HAP12 and AT130 to affect both nuclear cccDNAtranscription and cytoplasmic capsid assembly Cp functions.

Methods: HAP12 and AT130 effects on capsid-associated HBV-DNA, cccDNA and pgRNA levels (quantitative real-time PCR withspecific primers) were assessed in: (a) HBV-infected NTCP-HepG2cells; (b) AD38 inducible HBV stable cell line. Recruitment of HBcand histone modifications on the viral minichromosome wereassessed using the cccDNA ChIP assay in AD38 cells.

Results: CaMPs treatments resulted in a very strong inhibition ofHBV replication (>95%) and a significant but incomplete reductionof the stable cccDNA pool. A strong effect on cccDNA-dependentHBeAg production (AD38 tet-off) and pgRNA transcription (AD38tet-off/tet-on and NTCP-HepG2 infected cells) was also demon-strated. The ability of HAP12 to target cccDNA transcription wasconfirmed by the reduced cccDNA-bound H3 histone acetylationand the decreased HBc occupancy on the cccDNA in induced AD38cells. Importantly, when CaMPs treatment was started during infec-tion, cccDNA formation/accumulation was completely inhibited(>95%) and viral replication was blunted.

Conclusions: Anti-capsid compounds (CpAMs) have an impacton Cp nuclear functions at multiple levels: block of new cccDNAformation/accumulation, reduction of an established cccDNA pool


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C-21

ESIDUAL HCV-RNA IN LIVER EXPLANTS FROMATIENTS UNDERGOING SOFOSBUVIR ANDIBAVIRIN TREATMENT WHILE AWAITING LIVERRANSPLANTATION IS NOT ASSOCIATED WITHCV RELAPSE

. Gambato 1,2, S. Pérez-del-Pulgar 1,. Hedskog 3, J. Svarovskia 3, M.S. Paulson 3,

. Denning 3, M.P. Curry 4, N. Caro-Pérez 1,.C. Londono 1, X. Forns 1

Liver Unit, Hospital Clinic, IDIBAPS, CIBERehd,arcelona, SpainMultivisceral Transplant Unit, Department ofurgery, Oncology and Gastroenterology, Paduaniversity Hospital, Padua, ItalyGilead Sciences Inc., Foster City, CA, USABeth Israel Deaconess Medical Center, Boston, MA,SA

Background and aims: Sofosbuvir (SovaldiTM, SOF) in combi-ation with ribavirin (RBV) administered to HCV-infected patientsith hepatocellular carcinoma awaiting liver transplant (LT) pre-

ented recurrence of HCV post-LT in 70% in those individuals withndetectable serum HCV-RNA at time of LT (Curry et al., Gas-roenterology, 2014). The present study examined the presence ofCV-RNA in available liver explants from patients in the Phase 2linical study.

Patients and methods: Liver explant samples were collectedrom 38 HCV-infected cirrhotic patients that underwent LT. Allatients received 3–52 weeks of SOF + RBV while awaiting LT. HCV-NA levels in the liver were determined by quantitative real-timeCR in triplicate experiments. Liver explants from 39 HCV-RNAegative patients were used as controls.

Results: Thirty-four patients were included in the final analysis.wenty-three out of 34 (68%) patient liver explants were HCV-RNAositive (range 0.7–67566 copies/�g) and 11 (32%) were HCV-RNAegative. HCV-RNA was not detected in any of the control explants.reatment duration and time with undetectable HCV-RNA in serumefore LT were significantly lower in patients with HCV-RNA in

iver explants (16 and 7 weeks, respectively), compared to thoseith undetectable HCV-RNA (24 and 13 weeks, p = 0.037, p = 0.045,

espectively). Twenty-four out of 34 patients (71%) achieved SVR12fter LT and 10 patients (29%) presented recurrent HCV infection.nterestingly, HCV-RNA was detected in liver explants from 1668%) of the 24 responders and in 7 (70%) of the 10 non-respondersp = 0.850).

Conclusions: SOF + RBV regimen is an efficacious therapy forreventing HCV recurrence. The presence of HCV-RNA in liverxplants did not significantly correlate with the likelihood of recur-ent HCV infection post-LT. Residual HCV in liver explants mayepresent non-functional HCV-RNA (incomplete genomes versus
nfit viral strains) in the context of HCV-RNA polymerase inhibitiony SOF.

isease 47S (2015) e1–e18 e11

OC-22

A HYPER-GLYCOSYLATION OF HBV SURFACEMAJOR HYDROPHILIC REGION CORRELATESWITH IMMUNOSUPPRESSION-DRIVEN HBVREACTIVATION AND HAMPERS HBSAGRECOGNITION IN VITRO

L. Colagrossi 1, R. Salpini 1, M. Surdo 1, A. Battisti 1,M. Pollicita 1, A. Bertoli 1, F. Di Santo 1, C. Becker 2,C. Mastroianni 3, M. Marignani 4, S. Maylin 5,C. Delaugerre 5, F. Morisco 6, N. Coppola 7,A. Marrone 8, L. Sarmati 9, M. Andreoni 9,M. Angelico 10, J. Verheyen 11, C.F. Perno 1,V. Svicher 1

1 Department of Experimental Medicine and Surgery,Tor Vergata University, Rome, Italy2 Institute of Virology, University of Cologne,Germany3 “Sapienza” University, Rome, Italy4 Department of Gastroenterology, S.AndreaHospital, Rome, Italy5 Laboratoire de Virologie, AP-HP HopitalSaint-Louis, Paris, France6 Department of Clinical Medicine and Surgery,University of Naples Federico II, Section of InfectiousDiseases, Naples, Italy7 Department of Mental Health and Public Medicine,Section of Infectious Diseases, Second University ofNaples, Naples, Italy8 Internal Medicine and Hepatology Unit, SecondUniversity of Naples, Naples, Italy9 Infectious Diseases Unit, Tor Vergata UniversityHospital, Rome, Italy10 Hepatology Unit, Tor Vergata University Hospital,Rome, Italy11 Robert Koch Institute of Virology, University ofEssen, Essen, Germany

Introduction/aim: To investigate N-linked glycosylation pat-terns of HBsAg in immunosuppression-driven HBV-reactivationand to evaluate their impact on HBsAg-antigenicity.

Methods: Mutations associated with acquisition of N-glycosylation site were investigated in 127 HBsAg genotype-Dsequences from 47 patients with immunosuppression-drivenHBV-reactivation (defined as Hwang, 2014), and 80 chronicallyHBV-infected drug-naïve patients as control.

The impact of N-glycosylation sites on HBsAg-antigenicitywas analyzed by transfecting HepG2-cells with a plasmid encod-ing wild-type and mutated HBsAg linked to a streptavidin-tag(strep-tag). The strep-tagged HBsAg amount in supernatants wasquantified by a specifically designed ELISA targeting the Strep-tag(thus, not affected by HBsAg-mutations), and by ELISAs targetingthe HBsAg (Architect-Abbott, Monolisa-Biorad).

Results: Additional N-glycosylation sites are found in 19.1%(9/47) of HBV-reactivated patients versus 0/80 controls (P < 0.001).They localize in the major hydrophilic HBsAg-region (MHR), tar-get of antibodies. In 7 patients, a single additional N-glycosylationsite results from the mutations T115N (n = 2), T123N (n = 2),T131N (n = 2), and from the insertion of an N between 114 and115 position (ins114–115N) (n = 1). In the remaining 2 patients,2 additional glycosylation sites result from S113N+T131N and
ins114–115N+T117N, respectively.
Notably, 5/9 patients with ≥1 additional N-glycosylation sitesremain HBsAg-negative by diagnostic-test at HBV-reactivation(P = 0.002).

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In vitro, all additional N-glycosylation sites decrease thetrep-tagged HBsAg quantification by the 2 ELISAs targeting theBsAg. In particular, T115N, T123N, ins114–115N determine a90% decrease in HBsAg-quantification by both ELISAs. Similarly,ns114–115N+T117N cause a 90.2% and 75.4% reduction in HBsAg-uantification, respectively. No decrease of strep-tagged HBsAg

s revealed by ELISA targeting the Strep-tag. This suggests thatdditional N-glycosylation sites hamper HBsAg-recognition byntibodies without affecting HBsAg-release.

Conclusions: Additional N-glycosylation sites in MHR correlateith false HBsAg-negativity at ELISA despite HBV-reactivation, androfoundly affect HBsAg-antigenicity in vitro. This supports the rolef immune-escape mutations in HBV-reactivation during immune-uppression and the importance of HBV-DNA (more than HBsAg)n HBV-reactivation diagnosis.


C-23

BSAG LOSS IS ENOUGH TO DISCONTINUEONG-TERM NUCLEOS(T)IDE ANALOGUEHERAPY IN HBEAG-NEGATIVE CHRONICEPATITIS B PATIENTS IN REAL PRACTICE?

. Fasano 1, M. Ciarallo 1, G. Niro 2, R. Fontana 2,. Cozzolongo 3, A. Maci 4, I. Carraturo 4,. Miglietta 4, G. Angarano 5, T. Santantonio 1

Infectious Diseases, University of Foggia, ItalyGastroenterology, Casa Sollievo Sofferenzaospital, San Giovanni Rotondo (FG), ItalyDivision of Gastroenterology 1, National Institute ofastroenterology S. de Bellis, Castellana Grotte, Bari,

talyInfectious Diseases, Vito Fazzi Hospital, Lecce, ItalyInfectious Diseases, University of Bari, Italy

Introduction: Currently, oral nucleos(t)ide analogues (NAs) arehe most common therapeutic strategy for treatment of chronicepatitis B (CHB). HBsAg loss is a well-established endpoint ofherapy, but it is still unclear whether HBsAg clearance alone isnough to stop long-term NA therapy or seroconversion to anti-HBss required.

Aim: To evaluate in clinical practice the outcome in HBeAg-egative CHB patients who discontinued NA therapy after HBsAg

oss.Methods: HBeAg-negative CHB patients with HBsAg clearance

uring long-term NA therapy followed for at least 24 weeks after NAiscontinuation were retrospectively examined. HBsAg, anti-HBs,BV DNA levels and alanine aminotransferase levels were moni-

ored every 1–3 months after NA discontinuation in the first yearnd every 6 months thereafter. Virological, biochemical and clinicalata were collected and entered into a database.

Results: Among 590 monoinfected HBeAg-negative CHBatients who received NA therapy between 1998 and 2009, 244%) lost HBsAg after a median duration of therapy of 103 monthsrange 22–180) and were included in this study. Median age was 53ears (range 34–170), 18 were males, 18% cirrhotics. Twenty-twoatients received NA monotherapy (15 lamivudine, 3 entecavir, 4enofovir) and two patients received lamivudine + adefovir com-ination therapy. Only 11/24 patients (46%) developed anti-HBsanti-HBs level: ≤100 IU/ml in 5 and 100–500 IU/ml in 6), whereas3 patients remained persistently anti-HBs negative during follow-
p. All 24 patients stopped NA treatment after a median durationf consolidation therapy of 12 months (range 0–70). No case of
isease 47S (2015) e1–e18

virological and biochemical breakthrough were reported during 24months (range 6–180) of post-treatment follow-up.

Conclusion: HBsAg loss is a rare event in HBeAg-negative CHBpatients treated with NAs. However, regardless of anti-HBs sero-conversion and duration of consolidation therapy, long-term NAtherapy can be safely discontinued in clinical practice in patientswhit HBsAg loss, even in those with advanced liver disease.


OC-24

HIGH CAPABILITY OF CONTRAST ENHANCEDULTRASOUND IN DEFINING A RAPID DIAGNOSTICAND THERAPEUTIC WORK-UP FOR NODULES<2 CM IN CIRRHOTICS DURING SURVEILLANCE

A. Giorgio 1, G. Iaquinto 2, L. Montesarchio 1,P. Gatti 3, B. Santoro 4, F. Amendola 5,P. Matteucci 6, C. Coppola 2,7, V. Giorgio 7,8

1 Interventional Ultrasound Unit, Tortorella ClinicalInstitute, Consorzio ISMESS, Salerno, Italy2 Unità Operativa di Medicina Interna, Casa di CuraS. Rita, Atripalda, Avellino, Italy3 Unità Operativa di Medicina Interna, Ospedale diFasano, Brindisi, Italy4 Unità operativa ecografia interventistica, Athena,Caserta, Italy5 Unità Operativa di Medicina Interna, TortorellaClinical Institute, Consorzio ISMESS, Salerno, Italy6 Campus biomedico, Roma, Italy7 Unità Operativa di Medicina Interna, Epatologiainterventistica, ospedale di Gragnano, Napoli, Italy8 Università Cattolica del Sacro Cuore, PoliclinicoGemelli, Roma, Italy

Introduction: The disappearance of portal blood flow and thearterial vascularization is the hallmark of hepatocarcinogenesis,leading from benign regenerating nodules to well differentiatedHCC. The capability of a dynamic imaging modality detecting arte-rial hypervascurarization of small nodules is crucial to promote arapid diagnostic and therapeutic work-up improving survival.

Aim: To evaluate the capability of CEUS to detect arterial vas-cularization of ≤2 cm HCC nodules arising during surveillanceshorting diagnostic and therapeutic work-up.

Materials and methods: From July 2011 to June 2014 among1757 consecutive cirrhotics under surveillance with Ultrasound(US), 229 new single nodules 7–20 mm in diameter were detected.All these patients underwent CEUS followed by US guided percuta-neous 18 G needle core biopsy of the nodules as gold standard. Ofthe 229 nodules, 27 were hyperechoic lesions, 171 hypoechoic and31 isoechoic.

Results: On histology, 199/229 nodules were HCC and 30 werebenign. Of the 199 HCCs, CEUS showed arterial hypervascularityin 190 nodules (95.5%) [sensitivity 94.48%; specificity 100%; PPV100%, NPV 76.92%]. Of the 39 CEUS arterial-unenhanced nodules,30 were benign and 9 (4,5%) were well-differentiated HCC.

The fate of the 9 arterial-unenhanced CEUS HCC patients wasalso compared with that of the 6 HCC patients with no arterialenhancement on ultrafast dynamic gadolinium-enhanced MRI per-formed for staging in all HCC patients included in the study. Allthese 15 patients were treated with percutaneous radiofrequencyablation: 1-year distant recurrence rate was 11.1% in CEUS group
and 30% in MRI group (p < 0.01).
Conclusions: CEUS has a great capability to detect arterialhypervascularity of small HCC. Because only 4.5% of new nodules

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scape the demonstration of arterial hypervascularity, CEUS muste performed immediately after conventional US to contrast thealignant fate of small lesions arising in cirrhotics.


C-25

IR-17/92 EXPRESSION PATTERN: AOLECULAR SIGNATURE OF HCV-RELATEDIXED CRYOGLOBULINEMIA

. Piluso ∗, L. Gragnani, A. Genovesi, E. Fognani,. Monti, T. Urraro, A.L. Zignego

Department of Experimental and Clinical Medicine,niversity of Florence, Florence, Italy

Introduction: HCV infection is closely related to the devel-pment of lymphoproliferative disorders (LPDs), mainly mixedryoglobulinemia (MC) and B-cell lymphoma. Modification of thexpression levels of specific microRNAs (miRNAs) has been asso-iated with different autoimmune and/or LPDs. In particular, thendogenous miR-17/92 cluster is very often amplified in cancernd in autoimmunity. Scarce data exist about the modifications ofiRNA expression levels in HCV-related LPDs.Aim: To evaluate the modifications of miR-17/92 cluster levels

n HCV-positive patients with and without MC.Methods: miR-17/92 cluster expression levels were evaluated

y Real Time PCR in PBMC samples from 79 HCV patients: 34ithout LPD [HCV] and 45 with MC [HCV-MC]; among the 45C patients were included 20 patients who experienced a sus-

ained virological and clinical response after antiviral treatmentnd of which pre and post therapy sampling was available. Rela-ive expression levels of all the members of the miR-17/92 clusternamely, miR-17, miR-18a, miR-19a, miR-19b, miR-20a and miR-2a) were evaluated with the 2−��Ct method, using miR-let-7ds housekeeping.

Results: All the members of the miR-17/92 cluster were highlypregulated in PBMCs from HCV-MC patients (p < 0.001) whenompared to HCV group. A restoration of miRNAs levels wasbserved in the samples taken after viral eradication (miR-17, miR-8a, miR-19a, miR-19b, miR-20a, p < 0.001 and miR-92a, p < 0.05,hen compared with pre-treatment levels). Regarding miR-20a,

he levels in samples taken after HCV eradication continued to beignificantly higher than in controls (HCV patients) (p < 0.05), inpite of the sudden fall observed after therapy.

Conclusions: This study shows that the pattern of miRNA-17/92luster is modified in PBMC from HCV patients with MC. The suddenestoration of these values of expression in patients achieving austained virological and clinical response after antiviral treatment,
trongly suggests that miR-17/92 cluster plays a key role in theathogenesis of MC.

isease 47S (2015) e1–e18 e13

OC-26

THE ITALIAN ENTAS COHORT STUDY: ENTECAVIREFFECTIVENESS IN NAïVE AND TREATMENTEXPERIENCED PATIENTS WITH CHRONICHEPATITIS B

E. Porro 1, A. Di Leo 2, A. Marzano 3,G. Brancaccio 4, S. Maimone 5, M. Fasano 6,A. Grasso 7, F. Bronte 8, S. Fagiuoli 9,T. Santantonio 6, F. Morisco 10, E. Petrelli 11,G. Surace 12, G. Labbadia 13, L. Badia 14,G.A. Niro 15, M. Vinci 16, A. Montineri 17,F. Vinelli 18, M. Massari 19, L. Nosotti 20,G. Galati 21, G. Missale 1,22, Entas Study Group

1 Infectious Diseases and Hepatology, AziendaOspedaliero-Universitaria di Parma, Parma, Italy2 Gastroenterology and Digestive Endoscopy,University of Bari, Bari, Italy3 Gastroepatologia, Ospedale San Giovanni Battista,Torino, Italy4 Epatiti Virali Acute e Croniche, II Università degliStudi di Napoli, Napoli, Italy5 Clinical and Molecular Hepatology, UniversityHospital of Messina, Messina, Italy6 Malattie Infettive, Università degli Studi di Foggia,Foggia, Italy7 Gastroenterologia Osp San Paolo, Savona, Italy8 Gastroenterologia, Di.Bi.M.I.S., University ofPalermo, Palermo, Italy9 Gastroenterologia ed Epatologia Dei Trapianti,Ospedali Riuniti di Bergamo, Bergamo, Italy10 Gastroenterology Unit, Department of ClinicalMedicine and Surgery, University of Naples, FedericoII, Napoli, Italy11 Malattie Infettive, Azienda Ospedaliera SanSalvatore, Pesaro, Italy12 Clinica di Gastroenterologia, UniversitàPolitecnica delle Marche, Ancona, Italy13 Dipartimento di Medicina Interna e SpecialitàMediche, UOC Medicina Interna F, Sapienza,Università di Roma, Umberto I Policlinico, Roma,Italy14 Istituto Malattie Infettive, Azienda OspedalieroUniversitaria Policlinico S.Orsola Malpighi, Bologna,Italy15 Gastroenterology, Casa Sollievo SofferenzaHospital, IRCCS, San Giovanni Rotondo, Italy16 SC Epatologia e Gastroenterologia AO OspedaleNiguarda Cà Granda, Milano, Italy17 Malattie Infettive, PO Ferrarotto, Catania, Italy18 Gastroenterologia ed Endoscopia Digestiva,Azienda Ospedaliero Universitaria di Foggia, Foggia,Italy19 Infectious Diseases, IRCCS, Azienda Ospedaliera S.Maria Nuova, Reggio Emilia, Italy20 National Institute for Health, Migration andPoverty (NIHMP), Italy21 Unità di Medicina Clinica-Epatologia, PoliclinicoUniversitario Campus Bio Medico, Italy22 AISF Associazione Italiana per lo Studio del Fegato,Roma, Italy

Introduction: Real-life studies, mainly report results inuntreated patients.

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Aims: To define Entecavir antiviral potency and efficacy inntreated and IFN- and NUC-experienced patients in clinicalractice.

Material and methods results: 423 patients were evaluated forndetectable HBV-DNA, HBeAg seroconversion and monitored for

iver function and main complications.304 patients were untreated (naive), 97 IFN- and 34 NUC-

xperienced (12 IFN-experienced). Forty-five percent had liverirrhosis. Rate of undetectable HBV-DNA was 85–90% at 24–30onths in the whole cohort, while 84–90%, 87–91% and 85–80%

n naive, IFN- and NUC-experienced. Nine patients had been previ-usly treated with LAM or ADV and were not on treatment while 25witched to ETV or added ETV at enrollment. Two patients switchedo TDF, one because of virologic failure (L180M, M204V, M250V)nd the second for persistence of low viremia (<3 log 10) a third sub-ect added TDF because of partial virologic response. Two patientsre still viremic, one not compliant and the second is on dialysisreatment.

Thirty of 56 HBeAg positive patients seroconverted to anti-HBe53.5%) and 27 lost HBsAg (6.5%).

Bilirubin > 2 mg/ml, albumin < 3.5 g/dl and INR > 1.7 was in 15%,2% and 8% of cases at baseline with significant improvement at4 months (bilirubin, p < 0.001; albumin and INR, p < 0.05). Ascitesesolved in 22 of 37 cases while developed in 14 cases; 6 varicealleedings were observed. HCC was present in 41 patients at base-

ine and 30 developed HCC during follow-up with an incidence of.5% per year; 80% of new HCC cases were cirrhotic and significantlylder (p < 0.0001), while viremia, HBeAg status, coinfection (HCV,DV), ALT levels and BMI were not statistically different compared

o the other patients.Conclusions: The study of Entecavir treatment in field-practice

onfirms its efficacy in suppressing HBV replication also inreatment-experienced patients. Analysis of HCC rate confirmsesults of other real-life studies.


C-27

OTCH4 AND MHC CLASS II POLYMORPHISMSONTRIBUTE TO HCV-RELATED BENIGN ANDALIGNANT LYMPHOPROLIFERATIVE DISEASES

. Piluso 1,∗, L. Gragnani 1, A. Genovesi 1,

. De Re 2, E. Fognani 1, M. Libra 3, A.L. Zignego 1

Department of Experimental and Clinical Medicine,niversity of Florence, Florence, ItalyBio-Proteomics Facility, Department ofranslational Research, CRO, National Cancernstitute, Aviano, ItalyDepartment of Biomedical Sciences, Section ofathology and Oncology, Laboratory of Translationalncology and Functional Genomics, University ofatania, Catania, Italy

Introduction: Hepatitis C virus (HCV) can cause lympho-roliferative disorders (LPDs). The most frequent one is mixedryoglobulinemia (MC), clinically benign, but evolving in about0% of cases into a non-Hodgkin’s lymphoma (NHL). Recently, aWAS allowed the discovery of an association of two SNPs on chro-osome 6 and HCV-related MC; the first one is an intronic SNP

rs2071286) of the NOTCH4 gene, the second one (rs9461776) isocated between HLA-DRB1 and HLA-DQA1 gene segments.Aim:
o define the contribution of rs2071286 and rs9461776 in the pre-isposition to develop HCV-related LPDs.
isease 47S (2015) e1–e18

Methods: rs2071286 and rs9461776 were determined in 438patients: 85 HCV patients without LPDs (HCV), 73 HCV patientswith circulating cryoglobulins (MC-HCV), 108 with HCV-associatedMC syndrome (MCS-HCV), 62 with HCV-related NHL (NHL-HCV)and 110 HCV-negative patients with NHL (NHL).

Results: Concerning rs2071286, significantly higher minorallele frequency (maf) was observed in all the cases respect to con-trols (MC-HCV, p = 0.035; MCS-HCV, p < 0.001; NHL-HCV, p = 0.001and NHL, p = 0.019). Comparing to HCV group, the odds-ratio asso-ciated with rs2071286 maf were: MC-HCV, OR 1.79; MCS-HCV, OR2.59; NHL-HCV, OR 2.6; NHL, OR 1.81.

Regarding rs9461776, the higher maf was observed in NHL-HCVand in NHL groups respect to HCV (p = 0.011 and p = 0.0283, respec-tively). The presence of minor allele of rs9461776 conferred an ORof 2.42 to NHL-HCV and an OR of 1.7 to NHL group, in respect tocontrols.

Conclusions: We confirm the previously demonstrated associ-ation between the two SNPs and HCV-related MC vasculitis, andshowed similar associations for HCV-related NHLs and, regardingrs2071286, also for patients with CGs but without symptoms. Fur-thermore, HCV-negative NHLs showed higher frequencies of thetwo minor alleles respect to controls, but lower compared to HCV-positive lymphomas. This suggests that HCV acts on a permissivegenetic substrate and confirms the virus contribution to the lym-phomagenesis.


OC-28

17�-ESTRADIOL INHIBITS HEPATITIS C (HCV)INFECTION VIA BINDING TO ITS RECEPTOR

P. Giarda, A. Magri, C.Z. Foglia, E. Boccato,E.M. Burlone, R. Minisini, E. Grossini, M. Pirisi

Department of Translational Medicine, Univertitàdel Piemonte Orientale, Novara, Italy

Background: Hormonal factors may play a role in control-ling hepatitis C virus infection (HCV). In fact, spontaneous HCVclearance is more common among premenopausal women than inmen and, when infection persists, histologic progression of chronichepatitis is slower in pre-menopausal women in comparison topostmenopausal women and men. Aim of our study has been toevaluate sex hormones as inhibitors of HCV replication.

Methods: Huh-7.5 cells infected with the JFH-1 virus wereexposed to one each among the following hormones: dehy-droepiandrosterone (DHEA), testosterone, progesterone and 17�-estradiol (the latter in the presence/absence of the estradiolreceptor antagonist fulvestrant). Based on the inhibition of viralreplication, dose–response curves covering the physiological rangewere established allowing calculation of IC50 values. In model A,hormones were added 3 h post-infection. In model B, Huh-7.5 cellswere incubated with hormones for 1 h before being infected; 3 hlater, the inoculum was replaced with fresh medium. In model C,a 16 h pre-incubation 17�-estradiol preceded replacement withfresh medium, followed by infection.

Results: Progesterone and testosterone showed no inhibitoryeffect on viral replication at the concentrations tested for bothmodel A or B. DHEA exhibited only a partial antiviral effect in bothmodel (39%). In contrast, in model A 17�-estradiol inhibited viralreplication up to 45%, allowing to estimate IC50 = 490 nM. Moreover,the inhibitory effect of 17�-estradiol reached 67% in model B and
64% in model C, with IC50 values = 140 and 160 nM, respectively.Although fulvestrant did not exhibit any appreciable effect on viral

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eplication, in its presence the inhibition exerted by 17�-estradiolas reverted in a dose-dependent manner.

Conclusions: In vitro, 17�-estradiol is able to block HCV infec-ion, likely by modulation of intracellular pathways followinginding to the estradiol receptor, whose activation leads to anntiviral state.


C-29

CQUIRED SPHEROCYTIC LIKE ANEMIAOMBINED WITH INEFFECTIVERYTHROPOIESIS SUSTAINS ANEMIA INATIENTS WITH CHRONIC HEPATITIS C

NFECTION RECEIVING TELAPREVIR OROCEPREVIR-BASED TRIPLE THERAPY

. Lupo 1, R. Russo 2,3, A. Iolascon 2,3, D. Ieluzzi 4,

. Toniutto 5, S. Piovesan 6,7, E. Raffetti 8,. Siciliano 1, A. Matte’ 1, F. Turrini 9, F. Donato 8,. Alberti 6, V. Zuliani 1, G. Fattovich 1,4,. De Franceschi 1

Department of Medicine, University of Verona,erona, ItalyDipartimento di Medicina Molecolare eiotecnologie Mediche, Università degli Studi diapoli Federico II, Napoli, ItalyCEINGE Biotecnologie Avanzate, Napoli, ItalyClinical Unit of Gastroenterology, Aziendaspedaliera Universitaria Integrata Verona, Verona,

talyDepartment of Medicine and Pathology Clinical andxperimental, Medical Liver Transplantation Unit,nternal Medicine, University of Udine, Udine, ItalyDepartment of Molecular Medicine, University ofadua, Padua, ItalyClinical Unit of General Medicine, Aziendaspedaliera Universitaria Integrata Padua, Padua,

talyInstitute of Hygiene, Epidemiology and Publicealth, University of Brescia, Brescia, ItalyDepartment of Oncology, University of Turin, Turin,

taly

Background and aims: The addition of protease inhibitors,oceprevir (BOC) or telaprevir (TVR), to peg-interferon and riba-irin (PR) increases the incidence of anemia. Although geneticariants in ITPA gene have been linked to the hemolytic anemianduced by PR, the mechanism sustaining the severe anemia duringriple therapy is still unknown.

Materials and methods: We studied 59 patients with chronicepatitis C: 29 treated with TVR/PR and 30 with BOC/PR. Patientsere examined at baseline, at week 4 (end of PR lead-in phase),

t 12, 16 and 24 weeks of treatment. In all patients we evaluatediochemical and hematological parameters, red cell index; while

n a subgroup, we carried out in vitro functional studies to dis-ect the mechanism(s) underlying anemia in triple therapy. IL28Brs12979860) and ITPA (rs1127354, rs7270101) polymorphismsere also analyzed.

Results: In chronic HCV patients on triple therapy, we foundn early acute normochromic normocytic hemolytic anemia (4–8eeks) followed by a late macrocytic hypo-regenerative ane-
ia with low reticulocyte count in response to the degree of
nemia (12–24 weeks). Although the beneficial effects of ITPA poly-orphisms impacted the early phase of anemia (4 weeks), the

isease 47S (2015) e1–e18 e15

functional studies on red cells revealed: (i) the presence of sphe-rocytes; (ii) increased osmotic fragility; (iii) changes in red cellmembrane protein composition; (iv) increased phosphorylationof �-adducin with reduced membrane-cytoskeleton stability; (v)increased release of erythroidmicroparticles.

Conclusions: Our data indicate that the bimodal pattern of ane-mia in chronic HCV patients on triple therapy might be induced byacquired spherocytic-like anemia as dominating component in theearly phase (4–8 weeks), followed by the appearance of hypore-generative macrocytic anemia (12–24 weeks), most likely relatedto the combination effects of PR and TVR or BOC on erythropoiesis.These data could guide strategy of anemia management duringtriple therapy.


OC-30

SOFOSBUVIR-BASED ALL-ORAL TREATMENT FORELDERLY CHRONIC HEPATITIS C PATIENTS: ACOST-EFFECTIVENESS ANALYSIS

A. Ciaccio 1, P.A. Cortesi 2, G. Bellelli 3, M. Rota 4,S. Okolicsanyi 1, M. Rota 1, L. Mantovani 2,G. Annoni 3, M. Strazzabosco 1,5

1 Department of Surgery and Translational Medicine,University of Milano-Bicocca, Milan, Italy2 Research Centre on Public Health (CESP), Universityof Milano-Bicocca, Milan, Italy3 Department of Health Sciences, Geriatric Medicine,University of Milano-Bicocca, Milan, Italy4 Department of Health Sciences, Centre ofBiostatistics for Clinical Epidemiology, University ofMilan-Bicocca, Milan, Italy5 Liver Center & Section of Digestive Diseases,Department of Internal Medicine, Yale UniversitySchool of Medicine, New Haven, CT, United States

Background and aim: A relevant proportion of patients affectedby chronic hepatitis C (CHC) is older than 65 years. Comorbidi-ties and a higher susceptibility to drugs toxicity have historicallylimited treatment in these patients. Recent approval of interferon-free regimens, characterized by high efficacy and limited toxicity,provides unprecedented chances for these patients to be cured.

The aim of this study is to assess cost-effectiveness, taking intoaccount the severity of liver disease, age, and the geriatric (frailty)status.

Methods: A semi-Markov model of CHC natural history wasbuilt. The study focuses on CHC patients older than 65 years, strat-ified according to liver fibrosis (METAVIR F3 and F4), age (65–85years old) and Fried’s frailty phenotype (not frail, pre-frail and frail)generating 30 simulated cohorts. Treatment with sofosbuvir plussimeprevir (SOF/SMV) versus no treatment was assessed for eachcohort. The model estimated costs, Life Years and Quality AdjustedLife Years (QALY), with a lifetime time horizon and the HealthSystem perspective. Results are presented as incremental cost-effectiveness ratios (ICERs) per QALY gained. Cost-effectivenesswas defined as an ICER under the 37,000D threshold.

Results: At each fibrosis score, ICER increased with age andfrailty index. Among F3 patients, ICER ranged from D13,934 in not-frail 65-years-old andD79,354 in frail 85-years-old patients. AmongF4 patients ICER ranged from D13,873 in not frail 65-years-old andD115,965 in frail 85-years-old patients. In both F3 and F4 cohorts
ICER was below D37,000/QALY up to age 80 in non-frail patients,up to age 75 in pre-frail patients, up to age 70 in frail patients.

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Conclusion: SOF/SMV treatment is cost-effective in most CHCatients older than 65 years, however a careful assessment of theatient geriatric status is mandatory. This cost-effectiveness anal-sis should promote a prospective clinical study to verify efficacynd side effects in elderly HCV patients.


C-31

UTCOME INDICATORS IN PRIMARYCLEROSING CHOLANGITIS: INTERIM ANALYSISF THE VALUE-BASED MEDICINE INEPATOLOGY STUDY

. Fabris 1,2, A. Ciaccio 3, S. Okolicsanyi 3, M. Rota 4,.A. Cortesi 5, M.R. Buonocore 3, M. Gemma 3,. Giani 6, L.S. Belli 7, S. Fagiuoli 6, L. Scalone 5,.G. Valsecchi 4, L.G. Mantovani 5,. Strazzabosco 2,3

Department of Molecular Medicine, University ofadua, Padua, ItalyLiver Center & Section of Digestive Diseases,epartment of Internal Medicine, Yale Universitychool of Medicine, New Haven, CT, United StatesDepartment of Surgery and Translational Medicine,niversity of Milano-Bicocca, Milan, ItalyDepartment of Health Sciences, Centre ofiostatistics for Clinical Epidemiology, University ofilano-Bicocca, Milan, ItalyResearch Centre on Public Health (CESP), Universityf Milan-Bicocca, Milan, ItalyDepartment of Gastroenterology, Papa GiovanniXIII Hospital, Bergamo, ItalyDepartment of Hepatology and Gastroenterology,iver Unit, Niguarda Hospital, Milan, Italy

Introduction: Primary sclerosing cholangitis (PSC) is an enig-atic disease with scarce therapeutic options. The clinicalanagement of PSC remains challenging and may benefit fromutcome Indicators (OI) to assess the quality of care.

Aims: This study aims to: (A) identify OIs for PSC, and (B) validateIs in a clinical context.

Methods: (A) A panel of experts generated a list of OIs by Delphiethod. (B) OIs with the highest RAND/UCLA score were tested in

n ongoing multicentric, prospective study (Value-based Medicinen Hepatology, VBMH).

Results: Five OIs were identified having the highest rating val-es and low disagreement indexes: annual rate of acute cholangitispisodes (OI#1); mortality rate for patients not yet listed for liverransplantation (OI#2); rate of quality of life improvement, mea-ured by EQ-VAS and EQ-5D (OI#3); number of patients died forholangiocarcinoma and colo-rectal carcinoma (OI#4); incidencend/or worsening of osteoporosis (OI#5). In the validation study,3 consecutive patients with PSC enrolled in 3 tertiary centres inorthern Italy were evaluated for a median 24-months follow-uperiod. For each OI, the following values were reported: (OI#1)umulative incidence of 5.2%, resulting in 0.029 cholangitis/patient;OI#2, OI#4) no patients died without being listed for transplanta-ion or because of cancer during study time; (OI#3) 38.9 and 19.4%f patients showed an improvement in EQ-VAS and EQ-5D param-ters, respectively; (OI#5) 3% of patients developed or worsenedsteoporosis.

Conclusions: Five OIs for PSC were identified reporting highonsensus. Albeit the study population is small (as in the case ofare diseases) and the follow-up time is short as compared to the

isease 47S (2015) e1–e18

long natural history of the disease, these OIs have proven to be easyto collect and to work appropriately. Therefore, they are suitable tobe extended to specialized centres involved in PSC management tofurther validate their clinical usefulness.


OC-32

HEPCIDIN RESISTANCE IN DYSMETABOLIC IRONOVERLOAD

R. Rametta 1, S. Pelusi 1, P. Dongiovanni 2,F. Iuculano 1, A.L. Fracanzani 1,2, S. Fargion 1,2,L. Valenti 1,2

1 Department of Pathophysiology andTransplantation, University of Milan, Milan, Italy2 Department of Internal Medicine, FondazioneIRCCS Cà Granda Ospedale Maggiore Policlinico,Milan, Italy

Background and aim: The dysmetabolic Iron Overload Syn-drome (DIOS) is defined by hyperferritinemia associated withsteatosis and insulin resistance, and is associated with increasedrisk of hepatic and cardiovascular disease. The pathogenesis of ironaccumulation during DIOS is still unclear. Aim was to characterizeiron metabolism by performing an oral iron tolerance test (OITT)in DIOS as compared to hereditary hemochromatosis (HH) andhealthy individuals.

Methods: In 17 healthy volunteers, 10 C282Y+/+ homozygousHH, and 13 DIOS patients negative for mutations in HFE and FPN1,we administered 65 mg ferrous-sulfate orally. All subjects had nor-mal ferritin (30–150 ng/ml, >4weeks after depletion in HH andDIOS), normal erythropoiesis and inflammatory indices. Ferritin,transferrin saturation (TS%), and the iron hormone hepcidin wereevaluated at baseline (T0), 4 (T1), 8 (T2) and 24 (T3) h. Hepcidin wasmeasured by a new generation ELISA kit (DRG).

Results: HH had increased TS% and lower hepcidin than controlsand DIOS at all time points (p < 0.05). Despite baseline levels werecomparable to those of controls, DIOS patients had both higherTS% and hepcidin levels at T1 and T3 (p < 0.05 for all). A hepcidinrelease index did not differ between controls and DIOS, while itwas reduced in HH. Conversely, a hepcidin resistance index wassimilar between HH and controls, while it was twofold higher inDIOS compared to both controls and HH (p = 0.0004 and p = 0.004,respectively).

Conclusions: Iron accumulation in DIOS is not secondary todefective production of the hormone hepcidin, but it seems ratherdue to resistance to the action of hepcidin in decreasing serumiron and intestinal iron absorption. Additional studies are requiredto clarify the underlying molecular mechanisms. Conversely, dataconfirmed that inadequate hepcidin release underpins iron accu-

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C-33

CUTE KIDNEY INJURY IN CIRRHOTIC PATIENTSNDERGOING CONTRAST-ENHANCEDOMPUTED TOMOGRAPHY

. Filomia 1, S. Maimone 1, C. Saitta 1, L. Visconti 2,. Caloggero 3, A. Bottari 3, A. Alibrandi 4,. Cacciola 1,5, G. Caccamo 1, S. Spinella 1,. Vadalà 1, C.G. Gambino 6, M.S. Franzè 6,. Gambino 6, G. Raimondo 1,5, G. Squadrito 1,5

Division of Clinical and Molecular Hepatology,niversity Hospital of Messina, Messina, ItalyDivision of Nephrology, University Hospital ofessina, Messina, ItalyDepartment of Biomedical Sciences andorphological and Functional Imaging, University ofessina, Messina, ItalyDepartment of Economical, Business andnvironmental Sciences and Quantitative Methods,niversity of Messina, Messina, ItalyDepartment of Clinical and Experimental Medicine,niversity Hospital of Messina, Messina, ItalyUniversity School of Medicine of Messina, Messina,

taly

Introduction: Contrast-induced acute kidney injury (CI-AKI)ollowing intravenous contrast medium administration is one ofhe most common causes of hospital-acquired acute kidney dam-ge.

Aim: To investigate the incidence and possible predisposing fac-ors of CI-AKI in cirrhotic patients undergoing contrast-enhancedomputed tomography (CECT).

Patients and methods: We analysed 393 consecutively hos-italized cirrhotic patients. We excluded patients with activeacterial infection, glomerular filtration rate (GFR) < 30 ml/min,ecently intake of antibiotics and anti-inflammatory drugs. CECTas performed in 249 patients (CECT-group) and was not in the

emaining 144 (control group). No differences for age, sex, liverisease aetiology, Child-Pugh (CP) and MELD scores were presentetween the two groups. GFR was estimated using the Modificationf Diet in Renal Disease (MDRD-6) formula. The contrast induced-ephropathy (CIN) and the AKI-network (AKIN) criteria were usedo assess CI-AKI development. Serum creatinine was evaluated justefore and 72 h after CECT examination and, analogously, it wasested in control group at admission and 72 h later.

Results: CI-AKI occurred more frequently in CECT compared toontrol group (22/249 vs 5/144, p < 0.05 according to AKIN criteria,nd 42/249 vs 14/144, p = 0.05 according to CIN criteria). The fol-owing variables were significantly associated with CI-AKI at the

ultivariate regression analysis: male sex (p < 0.0001, OR: 0.118,I: 0.04–0.34), lower GFR (p = 0.022, OR: 1.019, CI: 1.003–1.035),

ower serum sodium levels (p = 0.025, OR: 0.87, CI: 0.770–0.982).I-AKI occurred independently of the presence of diabetes, arterialypertension, CP and MELD scores. Three-months follow-up wasvailable in 16/22 patients who developed CI-AKI showing thaterum creatinine persisted abnormally elevated in 10/16 (62.5%)atients.

Conclusions: CECT is a risk factor for the development of CI-KI in hospitalized cirrhotic patients independently of the presencef known causes of chronic kidney injury and of the severity ofiver disease. In these patients CECT-associated kidney dysfunction
eems to persist over time.

isease 47S (2015) e1–e18 e17

OC-34

OBLITERATIVE PORTAL VENOPATHY IN THEABSENCE OF CLINICAL PORTAL HYPERTENSION:AN UNEXPLORED PLANET

M. Guido 1, S. Sarcognato 1, A. Sonzogni 2,M.G. Lucà 3, M. Senzolo 4, S. Fagiuoli 3,A. Ferrarese 4, M. Pizzi 1, L. Giacomelli 1,G. Colloredo 5

1 Surgical Pathology & Cytopathology Unit,Department of Medicine-DIMED, University ofPadua, Padua, Italy2 Pathology Department, Papa Giovanni XXIIIHospital, Bergamo, Italy3 Gastroenterology and Transplant Hepatology, PapaGiovanni XXIII Hospital, Bergamo, Italy4 Multivisceral Transplant Unit, Department ofSurgical and Gastroenterological Sciences, UniversityHospital of Padua, Padua, Italy5 Department of Internal Medicine, San PietroHospital, Ponte San Pietro (BG), Italy

Obliterative portal venopathy (OPV) is the primary histologicallesion associated with idiopathic non-cirrhotic portal hyperten-sion (INCPH). Data on OPV in the absence of portal hypertension(PH) are very scarce. This retrospective study aimed to assess theprevalence of OPV lesions in patients with long-lasting abnor-malities of liver function tests (LFTs) of unknown etiology, butno evidence of PH. Four-hundred-eighty-two cases were retro-spectively collected from the clinical and pathological databasesof three referral centers for liver diseases. Criteria for inclusioninvolved the absence of clinical signs of PH and any known cause ofabnormal LFTs, and the histological demonstration of OPV lesions.Histological findings were matched towards clinical and labora-tory features. OPV lesions were identified in 19.9% of cases. LFTsabnormalities were mild in most cases. Serum autoantibodiesturned positive in 54.2% of patients. Pro-thrombotic and autoim-mune disorders were highlighted in 5.9% and 12.5% of testedcases, respectively. Histologically, the most prevalent OPV-relatedchanges were aberrant vessels (96.9%) and portal angiomatosis(74.0%). Among other INCPH-associated lesions, the most fre-quent were sinusoidal dilatation (65.6%) and increased number ofparenchymal veins (56.3%). Nodular regenerative hyperplasia wasdetected in 9.6% of cases. Conclusions: OPV changes occur in aconsistent subgroup of patients with abnormal LFTs of unknownorigin, even in the absence of clinical PH. OPV lesions are frequentlyassociated with histological and clinical features commonly relatedwith INCPH, suggesting that patients may be in an early pre-

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C-35

NTICOAGULATION DOES NOT INCREASEORTAL HYPERTENSION RELATED BLEEDING,UT EXPOSES PATIENTS WITH CIRRHOSIS TO AIGH RISK OF MINOR HEMORRHAGES: RESULTSROM A COMPARATIVE COHORT STUDY

. La Mura 1,2,∗, S. Braham 3, F. Branchi 4,. Moia 3, A.L. Fracanzani 5, M. Colombo 1,

. Tripodi 3, M. Primignani 1

U.O. Gastroenterologia-1, IRCCS-Ca’ Granda,spedale Maggiore Policlinico, Università degli studii Milano, Milan, ItalyU.O. Medicina Interna, IRCCS-San Donato,ipartimento di Scienze Biomediche per la Salute,niversità degli studi di Milano, Milan, ItalyCentro Emofilia e Trombosi Angelo Bianchi Bonomi,

RCCS-Ca’ Granda, Ospedale Maggiore Policlinico,niversità degli studi di Milano, Milan, ItalyU.O. Gastroenterologia-2, IRCCS-Ca’ Granda,spedale Maggiore Policlinico, Università degli studii Milano, Milan, ItalyU.O. Medicina Interna, IRCCS-Ca’ Granda, Ospedaleaggiore Policlinico, Università degli studi diilano, Milan, Italy

Introduction: Anticoagulation with vitamin K antagonistsVKAs) is an effective and relatively safe therapy for patients withortal vein thrombosis (PVT).

Aim: Exploring the haemorrhagic risk of VKAs in relation withhe presence of cirrhosis.

Material and methods results: Retrospectively, we comparedhe VKAs-related bleeding risk in cirrhotic patients with de novoVT (PVT-cohort, n = 62) vs non-cirrhotic patients with a throm-oembolic event (TE-cohort, n = 160). Any bleeding during 4-yearsf follow-up or withdrawal of anticoagulation, was recorded.leeding-risk due to portal hypertension (PHT) in the PVT-cohortas compared with an independent series of cirrhotics with

HT unexposed to VKAs during follow-up (CH-cohort, n = 53).ajor bleeding episodes under anticoagulation were intracranial

r retroperitoneal events, fatal bleeding events, need of hospi-alization or transfusion, otherwise they were considered minorleedings. All patients with cirrhosis were prophylaxed for PHT-elated bleeding according to current guidelines.

TE-cohort and PVT-cohort were comparable for age, sex andime in therapeutic range (TTR) of the INR. The treatment with VKAsas longer for the TE-cohort (31.1 ± 16.9 vs 23.0 ± 16.2 months,= 0.001). Overall, 41 patients under anticoagulation experiencedbleeding episode (14 major/27 minor). The actuarial probabilityf major/minor bleedings was higher in PVT-cohort (23%/30%) thann the TE-cohort (6%/20%) (p < 0.001). However, the risk of upper-astro-intestinal bleeding in PVT-cohort (15%) was the same as inhe CH-cohort (13%) also adjusting for potential confounders, con-rming that VKAs do not increase the risk of bleeding due to PHT.inally, the exclusion of the upper-gastrointestinal bleeding in PVT-ohort led to a significant reduction of major bleedings accountableor VKAs, leaving a significant residual risk only for minor episodesp < 0.05).

Conclusions: VKAs expose patients with cirrhosis and PVT ton additional risk of minor bleedings. This should be taken intoccount in future clinical studies to ameliorate the benefit/risk ratio
f anticoagulation in this clinical setting.

isease 47S (2015) e1–e18

OC-36

DIAGNOSTIC ACCURACY OF LIVER AND SPLEENSTIFFNESS MEASUREMENT FOR PORTALHYPERTENSION USING BIDIMENSIONAL SHEARWEAVE ELASTOGRAPHY

H. Stefanescu 1,2, B. Procopet 3,4, G. Allegretti 1,A. Berzigotti 3, N. Gamal 1, F. Conti 1, D. Festi 1,P. Andreone 1, L. Bolondi 1, J. Bosch 3, F. Piscaglia 1

1 Department of Medical and Surgical Sciences,University of Bologna, Italy2 Hepatology Department, Regional Institute ofGastroenterology and Hepatology, ClujNapoca,Romania3 Liver Hemodynamic Unit, Hospital Clinic yProvincial, University of Barcelona, Spain4 3rd Medical Clinic, University of Medicine andPharmacy, ClujNapoca, Romania

Background: Liver and spleen stiffness measurement(LSM;SSM) are used for the noninvasive assessment of chronic liverdiseases and may identify clinically significant portal hypertension(CSPH). Besides transient elastography new methods are proposed,but their performance to assess PH and their reliability criteria areunknown. We aimed to assess diagnostic accuracy for CSPH and todetermine a set of reliability criteria for bidimensional shear waveelastography (2D-SWE).

Methods: 51 consecutive cirrhotic patients (61% M; 59.7 yrs)submitted to HVPG measurement were enrolled. All patientsunderwent LSM and SSM by 2D-SWE (Aixplorer, Supersonic-Imaging). Multiple recordings of LS and SS were performed; thefinal result was calculated as their median. Each measurement pro-vides mean stiffness within Region of Interest (ROI) and standarddeviation (SD). The median of individual SD and the interquartilerange (IQR) of measurements for each patient were also calculated.

Results: LSM was successful in 47 patients (92.2%), while SSMonly in 42(82.4%). The median number of valid measurements was5 (3–11) for LSM and 4 (1–10) for SSM.

Both LSM and SSM correlated well with HVPG (r = 0.663;p < 0.0001 and r = 0.622; p < 0.0001, respectively). Median LSM andSSM significantly differed in patients without/with CSPH (15.0 vs.26.1 kPa; p < 0.0001 and 26.0 vs. 39.3 kPa; p = 0.001, respectively).

LSM and SSM had good performance to predict CSPH:AUROC = 0.893; accuracy = 75.5% (34/45) and AUROC = 0.837; accu-racy = 65% (26/40).

Using IQR < 30% of median LSM as additional reliability criteria,the accuracy increased up to 85.2% (23/27 patients). No benefit wasobserved by increasing the threshold of valid measurements > 3, orby using stricter SD (<20% or <10% of median LSM).

IQR/median and SD/median SSM (both <0.3) raised the accuracyto 76.5% (13/17 patients) and 82.3% (14/17 patients), respectively.

Conclusion: LSM and SSM by 2D-SWE have a good applicabil-ity in patients with PH and acceptable diagnostic performance for

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bstracts of the 48th A.I.S.F. – Italian Association for the Study of theiver – Annual Meeting 2015: Poster sessions – THURSDAY

-01

DRENERGIC HYPERFUNCTION IS A KEYRIGGER OF SOLUTE-FREE WATER RETENTIONN ASCITIC CIRRHOSIS: VASOPRESSIN (ADH) ISOT THE ONLY AGENT TO BLAME

. Sansoè 1, M. Aragno 2, R. Mastrocola 2,. Rosina 1, M. Parola 2

Division of Gastroenterology, Gradenigo Hospital,urin, Turin, ItalyDepartment of Clinical and Biological Sciences,niversity of Turin, Turin, Italy

Introduction Adrenergic hyperfunction causes proximal tubu-ar fluid retention and reduces renal excretion of solute-free water,ut, in advanced cirrhosis, non-osmotic hypersecretion of vaso-ressin (ADH) is considered the cause of water retention andyponatremia. Aim. Since ADH V2 receptor antagonists are noteneficial in long-term treatment of ascites, we hypothesize thatater retention in experimental ascitic cirrhosis depends also on

drenergic hyperfunction.Methods Hormonal status, renal function and tubular free-

ater reabsorption (TFWR) were assessed in 6 groups of ratsith ascitic cirrhosis: rats with cirrhosis due to 13-week CCl4

dministration (group G1); cirrhotic rats receiving daily diuretics0.5 mg/kg furosemide plus 2 mg/kg K+-canrenoate) during 11th-3th weeks of CCl4 (G2); cirrhotic rats treated daily with diureticsssociated with guanfacine oral prodrug (�2A adrenergic recep-or agonist and sympatholytic agent) 2 (G3), 7 (G4), or 10 mg/kgG5), or SSP-004240F1 (V2 receptor antagonist) 1 mg/kg (G6) during1th-13th weeks of CCl4.

Results Sodium excretion was lower in G1 than G2-G4 and6 (all P < 0.05). TFWR was higher in G1 (untreated cirrho-is, 32 ± 11 microL/min) than in G6 (diuretics + V2-antagonists,1 ± 9 microL/min) or G3 (diuretics + guanfacine 2 mg/kg, 20 ± 8icroL/min) (all P < 0.03). Guanfacine, added in G3 to diuretics (G2),

educed serum norepinephrine from 423 ± 122 to 211 ± 111 ng/LP < 0.01), plasma renin activity from 25 ± 12 to 9 ± 7 ng/mL/hP < 0.03), and TFWR from 45 ± 18 to 20 ± 8 microL/min (P < 0.01).mong all rats, TFWR did not correlate with ADH levels (r = 0.02, P:
.s.), but did with plasma aldosterone (r = 0.51, P < 0.01) and urinaryotassium excretion (r = 0.90, P < 0.001).
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Conclusions In ascitic cirrhosis, reduced volaemia, adrenergichypertone, and secondary aldosteronism, especially when exac-erbated by furosemide, contribute to tubular water retention.Sympatholytic agents are as effective as V2-antagonists to blunttubular water retention.


T-02

CHRONIC PARTICULATE MATTER EXPOSITIONAND WESTERN DIET INDUCE THE PROGRESSIONFROM STEATOSIS TO STEATOHEPATITIS

M. Tarocchi 1,∗, G. Marroncini 1, S. Polvani 1,S. Tempesti 1, E. Ceni 1, T. Mello 1, M. Peluso 2,A. Galli 1

1 Department of Experimental and ClinicalBiomedical Sciences, University of Florence, Florence,Italy2 Cancer Risk Factor Branch, Cancer Prevention andResearch Institute, Florence, Italy

Introduction: Nonalcoholic fatty liver disease (NAFLD) is themost common chronic liver disease, and the prevalence is rapidlyincreasing in developed countries. Nonalcoholic steatohepatitis(NASH), the severe form of NAFLD, can also progress to liver cir-rhosis and hepatocellular carcinoma. Recent evidences suggest thatenvironmental factors can trigger hepatic inflammation and pro-gression of steatosis to NASH.

Aim: We evaluate if a western style diet in association withchronic urban particulate matter exposition can modifies thepathogenesis and progression of NASH.

Materials and methods: The experimental model was created toreproduce urban lifestyle: C57Bl/6 mice were fed with a westernstyle diet (HFD), and treated with particular matter (PM) collectedfrom the urban area of Florence (Italy). After 4 and 8 weeks wereperformed the morphologic analysis of liver tissues, the evaluationof inflammatory cell infiltrate and the collagen deposition; we eval-uate also the effects of PM on cytokine production and oxidativestress related cellular damage.

Results: Both the HFD groups developed fat accumulation in theliver, and at 8 weeks, the NASH score was significantly increasedin HFD-PM group (p < 0.01) based on the histological analysis,the tissue fat content, the inflammatory cell infiltrate and the

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ollagen deposition. The cytokine profiles indicate an increasedroduction of pro-inflammatory molecules in presence of PM. The

evels of DNA adducts were significantly increased in HFD groupsnd,interestingly, the effects of HFD and PM were synergistic in facthe DNA adducts were about thirty fold greater in HFD mice com-ared to controls treated with the same amounts of air particulateatters.Conclusions: Our data suggest that in subjects with steatosis

ue to a western style of life, the association with a chronic expo-ition to urban particulate matter plays an important role in theathogenesis of NASH and its evolution to cirrhosis and also toancer.


-03

NSULIN RESISTANCE AND LIVER DAMAGE ARESSOCIATED WITH EARLY SIGNS OF LEFTENTRICULAR SYSTOLIC DYSFUNCTION INON-DIABETIC, NON-DYSLIPIDEMIC,ORMOTENSIVE PATIENTS WITHONALCOHOLIC FATTY LIVER DISEASE

. Vanni 1,∗, L. Mezzabotta 1, R. Faletti 2,. Morello 3, A. Marengo 1, G. Battisti 2, S. Frea 3,. Cannillo 3, C. Rosso 1, E. Mosso 1,

. Bergamasco 4, M. Rizzetto 1, E. Bugianesi 1

Department of Medical Sciences, Division ofastroenterology, University of Turin, Turin, ItalyDepartment of Diagnostic Imaging andadiotherapy, Radiology, University of Turin, Turin,

talyDepartment of Medical Sciences, Division ofardiology, University of Turin, Turin, ItalyUniversity of Turin, Turin, Italy

Background andAims: Nonalcoholic Fatty Liver Disease (NAFLD)as been associated with subclinical cardiovascular disease (CVD).his study was undertaken to evaluate the relationship betweenetabolic parameters, histologic features and parameters of car-

iac morphology and function in NAFLD subjects.Methods: Nineteen non-diabetic, non-dyslipidemic, non-

ypertensive patients with biopsy-proven NAFLD (17 men, 41 ± 8ears, BMI 26.8 ± 3 kg/m2) and 9 healthy controls (5 men, 30 ± 2ears, BMI 22.5 ± 2 kg/m2) underwent transthoracic echocar-iography and cardiac-MRI to evaluate cardiac morphology andunction. Endogenous glucose production (EGP) and lipolysis weressessed by stable isotope tracers. Hepatic Insulin resistance (IR)s EGPxfasting insulin, Oral Glucose Insulin Sensitivity (OGIS),dipo-IR as free fatty acids (FFAs)xfasting insulin were calculated.

Results: NAFLD patients had significantly higher con-entration of FFAs than controls (p < 0.05) and higher totalaturated and monounsaturated levels (p < 0.05). In NAFLDasal Hepatic-IR (NAFLD vs controls: 92 ± 34 vs 52 ± 18mol/minkg*mU/L) and Adipo-IR (NAFLD vs controls: 21 ± 10s 11 ± 5 mmol/L*mU/L) were significantly increased (p < 0.03 forll) and OGIS significantly reduced (NAFLD vs controls: 11.0 ± 1.64s 13.1 ± 1.1 mg/kgmin, p = 0.005). The end-systolic LV diameter30.4 ± 3.7 vs 27.2 ± 3.5 mm, p = 0.044) was significantly highern patients than in controls. In NAFLD patients, both hepatic-IRnd adipo-IR directly correlated with MRI end systolic LV volumeESV) (r = 0.63, p = 0.004 and r = 0.54, p = 0.018, respectively), while
GIS was inversely related to end-systolic LV diameter (r = -0.48,= 0.037) and ESV (r = -0.48, p = 0.036). At liver biopsy, steatosis33% was associated with increased ESV (p = 0.047), suggesting
sease 47S (2015) e19–e42

early systolic dysfunction. Similarly, ESV was increased in patientswith fibrosis (69.2 ± 16.9 vs 94.5 ± 31.4cc, p = 0.018), whereasejection fraction (51 ± 7 vs 59 ± 7%, p = 0.034) and cardiac index(2.9 ± 0.7 vs 3.8 ± 0.9, p = 0.03) were significantly reduced.

Conclusions: In NAFLD subjects metabolic derangements andhistological features are associated with early systolic LV dysfunc-tion, independently of diabetes, hypertension and dyslipidemia.

Funded by FP7/2007-2013 under grant agreement n HEALTH-F2-2009-241762 for the project FLIP and by PRIN 2009ARYX4 T


T-04

NLRP3 INFLAMMASOME INCREASES HEPATICFIBROSIS BY INDUCING INFLAMMATORYSIGNALS IN HEPATIC STELLATE CELLS

C. Rychlicki ∗, L. Agostinelli, E. Mingarelli,S. Saccomanno, C. Pinto, I. Pierantonelli, L. Trozzi,A. Benedetti, M. Marzioni, S. De Minicis,G. Svegliati-Baroni

Department of Gastroenterology, PolytechnicUniversity of Marche, Ancona, Italy

Background and Aims: Hepatic fibrosis represents the wound-healing response process to chronic liver injury, independentlyfrom aetiology. Hepatic stellate cells (HSCs) are the main liver extra-cellular matrix producing cells and also exert proinflammatoryactivity. The NLRP3 inflammasome mediates the release of pro-inflammatory cytokines in response to cellular danger signals invarious organs, but its role in the progression of hepatic injury isstill unclear. Thus, aim of the study was to evaluate the role of NLRP3inflammasome in fibrosis development and in HSCs behaviour.

Methods: Wild-type (WT) C57BL/6 and Nlrp3A350VneoR(Nlrp3-/−)mice underwent bile duct ligation for 3 weeks. Quiescent HSCs(quHSCs) and in vitro activated HSCs (AcHSCs) from WT mice wereobtained respectively after 12 hours and 6 days of culture.

Results: Compared to WT, Nlrp3-/-BDL mice showed a significantreduction in liver inflammasome activation, associated to signifi-cantly decreased TLR4, TLR9 and IL6 mRNA levels. Type I collagen,TGF�, CTGF and TIMP1 gene expression were significantly reducedin Nlrp3-/-BDL mice, leading to reduced collagen deposition mea-sured by Sirius Red staining. In vitro HSCs activation was associatedto significantly decreased gene expression of inflammasome com-ponents compared to (qu)HSCs. However, (Ac)HSCs stimulationwith LPS induced expression of the inflammasome pathway, with-out affecting neither type I collagen mRNA nor cell proliferation.After priming with LPS, incubation with ATP was needed for therelease of IL1�. (Ac)HSCs incubation with recombinant IL1� fur-ther stimulated the inflammasome pathway and led to increasedTGF� and CTGF gene expression and HSCs migration.

Conclusions: NLRP3 inflammasome contributes to hepatic fibro-sis by increasing the inflammatory response and inducing aparacrine loop that maintains TLRs activation and IL6 production.IL1� released by (Ac)HSCs stimulates fibrogenesis and cell migra-tion in an autocrine manner. Several new therapeutic targets can

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-05

NOVEL ROLE FOR THE KYNURENINEATHWAY IN EXPERIMENTAL STEATOHEPATITIS

. Vivoli 1, A. Cappon 1, A. Cozzi 1, N. Navari 1,. Gargano 2, F. Fallarino 2, F. Marra 1

Dipartimento di Medicina Sperimentale e Clinica,niversity of Florence, Florence, ItalyMedicina Sperimentale, University of Perugia,erugia, Italy

Background and Aims: The pathogenic mechanisms underly-ng development of nonalcoholic steatohepatitis are still elusive.ndoleamine 2,3-dioxygenase (IDO-1), an enzyme that mediateshe catabolism of L-tryptophan to L-kynurenine, plays an impor-ant role in hepatic immune regulation, mediating inflammation orolerance depending on the type of injury and the tissue involved.n the present study, we examined the effect of pharmacological orenetic inhibition of IDO-1 on the development of steatohepatitisnduced by a methionine and choline-deficient (MCD) diet in mice.

Methods: Balb/C mice fed a MCD diet for 8 weeks, werereated with the specific IDO inhibitor, 1-methyl-D-triptophan1MT, 5 mg/ml) or its vehicle in drinking water. WT and IDO-1 KO57Bl/6 mice were fed MCD or its control for 4 weeks. Intrahepaticene expression was assayed by quantitative real time PCR.

Results: 1MT administration caused a significant reduction ofLT levels in MCD-fed mice, while no changes were observedomparing the WT + MCD and IDO-1 KO + MCD groups. Both phar-acologic and genetic interference with IDO-1 resulted in an

melioration of the inflammatory phenotype. These effects weressociated with reduced intrahepatic expression of proinflam-atory factors, including CD11b, CCL2, TNF�, and IL-1�. Fibrosis

nduced by the MCD diet was decreased by 1MT co-administrationperformed for 8 weeks), together with reduced intrahepatic genexpression of TGF-� and �-SMA.

Conclusions: Interference with the IDO pathway by 1-MTdministration or by genetic deletion of IDO-1 ameliorates the phe-otype of dietary experimental steatohepatitis and modulates thexpression of proinflammatory and profibrogenic factors in thisontext.


-06

EDUCTION IN SUMOYLATION-DEPENDENT100A4 NUCLEAR IMPORT INHOLANGIOCARCINOMA BY LOW DOSEACLITAXEL HALTS TUMOR INVASIVENESS ANDEMATOGENOOUS METASTASIZATION BYOWN-MODULATING RHO-A AND CDC42CTIVITIES

. Spagnuolo 1,∗, M. Cadamuro 1,2, L. Sambado 1,. Indraccolo 3, G. Nardo 3, A. Rosato 3, E. Novelli 4,. Spirli 5, M. Strazzabosco 1,5, L. Fabris 1,2,5

Dep. of Surgery & Translational Medicine,niversity of Milan-Bicocca, ItalyDep. of Molecular Medicine, University of Padua,

talyIstituto Oncologico Veneto, IRCCS, Padua, ItalyClinica San Gaudenzio, Novara, Italy
Section of Digestive Diseases, Yale University, USA
Background. Cholangiocarcinoma (CCA) is characterized byarly and strong invasiveness. Nuclear expression of the S100A4

sease 47S (2015) e19–e42 e21

protein is a marker of increased CCA invasiveness and our pre-liminary data showed that Paclitaxel (PTX) could reduce S100A4nuclearization. We aimed at studying if nuclear S100A4 promotesCAA invasiveness and may represent a therapeutic target.

Methods and Results. CCA cells expressing nuclear S100A4(EGI-1) were treated with increasing PTX doses to study its effectson nuclear S100A4 expression, S100A4 sumoylation (a mechanismof nuclear import of proteins), cytoskeletal integrity, cell prolifer-ation/apoptosis, motility, invasiveness and Rho GTPases activity.PTX (1.5 and 15 nM) induced a marked reduction in nuclear S100A4.This decrement was linked with a significantly reduced sumoylatedfraction and an attenuation of cell migration, invasiveness andRho-A/Cdc42 activation, without affecting proliferation/apoptosisor cytoskeletal integrity. SCID mice xenografted with EGI-1 cellsby spleen injection, were treated, after tumor engraftment, withlow-dose metronomic regimen of PTX (2,6 mg/kg/die; n = 5) for14 days, using untreated mice as controls (n = 5). After micesacrifice, we evaluated PTX effects on tumor size, number ofmicrometastasis (MM) and isolated tumour cells (ITC) in liverand lung samples, along with proliferation, apoptosis and S100A4expression of EGI-1. PTX induced a significant reduction in both thetumor size and number of lung MM/ITC. The primary CCA mass ofPTX-treated mice, showed a significantly reduced number of EGI-1cells expressing nuclear S100A4, whereas proliferation/apoptosisrate did not change.

Conclusion. Down-regulation of nuclear S100A4 by PTX at dosesbelow those commonly used in chemotherapy but able to interferewith the sumoylation process, results in a decreased CCA cell motil-ity and invasiveness and in a reduction of hematogenous spread.These effects are not dependent upon changes in cell proliferation,apoptosis or cytoskeleton integrity, but upon down-modulation ofRho-A and Cdc42 activities by a reduced S100A4 nuclearization.


T-07

COMPLICATIONS AFTER PERCUTANEOUSRADIOFREQUENCY ABLATION (RFA) OFHEPATOCELLULAR CARCINOMA (HCC) INCIRRHOSIS: 20 YEARS EXPERIENCE IN A SINGLECENTER

A. Giorgio 1,∗, G. Iaquinto 2, L. Montesarchio 1,P. Gatti 3, B. Santoro 4, F. Amendola 5,P. Matteucci 6, C. Coppola 2,7, V. Giorgio 7,8

1 Interventional Ultrasound Unit, Tortorella ClinicalInstitute, Consorzio ISMESS, Salerno, Italy2 Unità Operativa di Medicina Interna, Casa di CuraS. Rita, Atripalda, Avellino3 Unità Operativa di Medicina Interna, Ospedale diFasano, Brindisi, Italia4 Unità operativa ecografia interventistica, Athena,Caserta, Italia5 Unità Operativa di Medicina Interna, TortorellaClinical Institute, Consorzio ISMESS, Salerno, Italy6 Campus biomedico, Roma, Italia7 Unità Operativa di Medicina Interna, Epatologiainterventistica, ospedale di Gragnano, Napoli, Italia8 Università Cattolica del Sacro Cuore, PoliclinicoGemelli, Roma, Italia

Aim: To report 20 years experience on complications after RFA
of HCC in cirrhotics, giving special emphasis on the changes of ChildClass, haemorragic events and deaths.
Material and Methods: From April 1994 to March 2014, 1787RFA procedures were performed percutaneously under ultrasound

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uidance in 1162 consecutive cirrhotics (57-85 years; mean 68;82 males; 852 Child A, 310 Child B). Diameter of HCC nod-les ranged from 1.2 to 6.8 cm. ProtrombinTime (PT) and Plateletsount were > 50% and 50.000 mm3, respectively; total bilirubinanged from 0,80 and 3.4 mg/dl (mean 1.6). Three RFA devicesere employed: hooks elecrtrode-needle (170 cases), perfused

lectrode-needle (1041 cases) and cold electrode- needle (476ases). 67 cirrhotics underwent RFA on both intraparenchymal HCCodule and tumor thrombus in the main portal vein (PVTT).

Results: Mortality. 4 patients (0,4%) died after RFA: 2 foraemoperineum, 1 for haemotorax, 1 for liver failure. No patientreated for both intraparenchimal HCC and PVTT died.

Morbidity. Changes in Child Class of cirrhosis were observed in9 patients (1.6%): 8 Child A cirrhotics had development of liverecompesation (ascites and/or jaundice and/decreased PT) chang-

ng from Child A to Child B class in 7, and from Child A to Childin 1 case, respectively. 11 Child B patients changed to Child C

lass. 6 patients had haemoperitneum with no death. Abscess for-ation was observed in 2 patients. One patient had an intraepatic

aematoma, that resolved spontaneously.Conclusion: RFA of HCC in cirrhotic patients can be consid-

red safe, even in case of advanced disease, such as invasionf portal venous system. Care must be taken in the evaluationnd surveillance of functional liver reserve. Haemorragic adversevents remain the leading cause of mortality.


-08

HARMACOLOGICAL INHIBITION OF SIRT1ROMOTES APOPTOSIS AND SENESCENCE OFEPATIC STELLATE CELLS DURING LIVER INJURY

. Tarocchi ∗, G. Marroncini, E. Ceni, S. Polvani,. Tempesti, T. Mello, A. Galli

Department of Experimental and Clinical Biomedicalciences, University of Florence, Florence, Italy

Introduction: Hepatic fibrosis is characterized by excessiveeposition of extracellular matrix (ECM) which leads to alterations

n liver function. Activated hepatic stellate cells (HSC) are consider-te to be the key cell type responsible for excessive ECM depositionfter liver injury. Modulation of SIRT1, a NAD-dependent histoneeacetylase, seems to regulate cellular metabolism, senescence andpoptosis.

Aim: We investigated the role of SIRT1 activation in HSC duringiver injury as a possible therapeutic target for liver fibrosis.

Materials and methods: We evaluated the expression of SIRT1n human tissues; we evaluated its role in response to stress condi-ions in primary human and mouse HSC. We also tested the in vivoffects of SIRT1 inhibition in different mouse models of liver injurynduced by carbon tetrachloride administration and bile duct liga-ion.

Results: We found that SIRT1 is constitutively expressed in HSC,nd in higher levels compare to other hepatic cells. In vitro, weemonstrated that starving, oxidative stress or hypoxia resulted

n SIRT1 increased activity and protected HSC in stress conditions.onversely its pharmacological inhibition reduced HSC lifespan dueo higher susceptibility to pro-apoptotic stimuli and the acquisitionf a senescent phenotype. Selective SIRT1 inhibition in mouse mod-ls of liver injury lowered the activated HSC specific genes as aSMA,MPs, TIMPs and pro-inflammatory cytokines. This was mainly

ue to a decrease in activated HSC content in the parenchyma. Inter-stingly this alteration in the hepatic cellular composition resultedn a reduction in collagen deposition.

sease 47S (2015) e19–e42

Conclusions: Selective SIRT1 inhibition in vitro increases stress-related senescence and apoptosis in primary HSC; the modulationof SIRT1 in the animal model liver decreases the number of acti-vated and proliferating HSC with a consequent reduction of ECMdeposition. The inhibition of SIRT1 could therefore prevent thefibrogenic alteration of the parenchyma during liver injury andpromote the recovery process.


T-09

JNK SIGNALING ACTIVATED BYPLATELET-DERIVED GROWTH FACTOR D(PDGF-D) STIMULATES SECRETION OF VASCULARENDOTHELIAL GROWTH FACTOR-C (VEGF-C) BYCANCER-ASSOCIATED FIBROBLASTS TOPROMOTE LYMPHANGIOGENESIS AND EARLYMETASTATIZATION IN CHOLANGIOCARCINOMA

M. Cadamuro 1,2, M. Vismara 1, S. Brivio 1,A. Furlanetto 3, M. Strazzabosco 1,4, L. Fabris 1,2,4

1 Department of Surgery and Translational Medicine,University of Milan-Bicocca, Milan, Italy2 Department of Molecular Medicine, University ofPadua, Padua, Italy3 Pathology Unit, Treviso Regional Hospital, Italy4 Section of Digestive Diseases, Yale UniversitySchool of Medicine, New Haven, Connecticut, USA

Background: Cholangiocarcinoma (CCA) is a highly invasivemalignancy with a poor prognosis. Currently, < 30% of CCA patientsundergo surgical resection, due to the early lymph node metasta-sization. In CCA, lymphangiogenesis develops within an abundantstroma, mainly composed by cancer-associated fibroblasts (CAF)recruited by Platelet-derived Growth Factor-D (PDGF-D) secretedby CCA cells. Mechanisms governing lymphangiogenesis in CCA areunknown. We aimed at investigating the role of PDGF-D-mediatedepithelial-mesenchymal interactions in promoting lymphangio-genesis.

Methods/Results: Immunohistochemistry on human CCA spec-imens (n = 6), were performed to understand the relationshipamong lymphatic vessels (D2-40+), blood vessels (CD34+), CCA andCAF (�SMA+), and the expression of PDGF-D, PDGFR�, VEGF-C andVEGFR3. Furthermore, lymphatic (LMVD) and vascular microves-sel density (VMVD) were calculated in CCA and compared withhepatoma (n = 6). With respect to hepatoma, CCA was charac-terized by an increased LMVD and a reduced VMVD. In CCA,lymphatic endothelial cells (LEC) laid in close vicinity to CAF andto cancer cells. We observed a specific expression of PDGF-D (onCCA cells), PDGFR� and VEGF-C (CAF), and VEGFR-3 (LEC), sug-gesting a sequential CCA cells-CAF-LEC crosstalk. To verify this,we measured in primary fibroblasts challenged with PDGF-D thesecretion of lymphangiogenic growth factors (VEGF-C, Ang-1 andAng-2) (ELISA). PDGF-D stimulated fibroblasts to secrete VEGF-Cbut not Ang-1 (Ang-2 was not expressed), an effect significantlyreduced by PDGFR� (imatinib mesylate), and JNK (SP600126)inhibitors. Using Boyden chambers, we evaluated the motilityof human LEC following stimulation with conditioned mediumof PDGF-D-challenged fibroblasts with/without SP600126. Con-ditioned medium induced a significantly stronger effect on LECrecruitment than that of unstimulated fibroblasts, an effect signif-icantly hampered by SP600126.

Conclusions: PDGF-D released by CCA cells stimulates CAF tosecrete VEGF-C through JNK signaling. In turn, VEGF-C secreted by

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-10

HE AMPK RELATED KINASE NUAK2 INTERACTSITH TGF-BETA AND REGULATES THE

CTIVATION PROCESS OF HEPATIC STELLATEELLS (HSC)

. Provenzano 1,∗, C. Tosti Guerra 1, A. Caligiuri 1,

. Rombouts 2, M. Pinzani 2, F. Marra 1

Dipartimento di Medicina Sperimental e Clinica,niversity of Florence, Florence, ItalyInstitute for Liver and Digestive Health, Universityollege London, London, United Kingdom

Background and Aims: Nuak2 is a member of AMPK relatedinases (ARKs), that act as energy sensors and controllers of cellulartructure, with different effects on cell motility and cytoskeletalrganization, depending on the cell types. AMPK possesses anti-brogenic activity. A recent study showed a link between Nuak2nd TGF�, a major driver of hepatic fibrogenesis-. However, littlenformation is available on the role of Nuak2 in liver fibrosis, andn particular on HSC trans-differentiation. Aims of this study waso elucidate the involvement of Nuak2 in HSC transactivation, ando investigate the possible interaction between Nuak2 and TGF�ignaling.

Methods: HSC were isolated from normal rat and human livernd activated by culture on plastic. Knockdown of Nuak2 waschieved by siRNA. Cell migration was evaluated in modified Boy-en Chambers. Protein expression and signaling pathways werenalyzed by Western blotting.

Results: In fully activated HSC, down-regulation of Nuak2ositively modulated cell migration and induced changes in thexpression of molecules involved in cytoskeletal organization.nockdown of Nuak2 increased �-SMA and p-SMAD3 expression,

n HSCV exposed to TGF�. Moreover, Nuak2 was up-regulated inSC following TGF� treatment.

Conclusions: The AMPK related kinase, Nuak2, is modulateduring the activation process of HSC, regulates cytoskeletal organi-
ation and cell motility and interacts with TGF-� in regulating thero-fibrogenic properties of HSC.

sease 47S (2015) e19–e42 e23

T-11

THE ITA.LI.CA STAGING SYSTEM FOR PATIENTSWITH HEPATOCELLULAR CARCINOMA: AMULTICENTER COHORT STUDY

F. Farinati 1,1, A. Vitale 1,1, F. Trevisani 2, T. Huo 3,4,Y.-H. Lee 3,4, U. Cillo 1, On behalf of the ITA.LI.CA.study group

1 Department of Surgery, Oncology andGastroenterology, University of Padua, Padua, Italy2 Department of Medical and Surgical Sciences,Division of Semeiotics, Alma MaterStudiorum–University of Bologna, Bologna, Italy3 National Yang-Ming University, Taiwan4 Department of Medicine, Taipei Veterans GeneralHospital, Taipei, Taiwan

Background The Barcelona Clinic Liver Cancer (BCLC) clas-sification ability to stratify HCC patient survival still remainscontroversial when compared to that of other systems (HKLC,MESIAH, CLIP, JIS). The aim of this study is to develop and validatea new staging system for HCC.

Methods The ITA.LI.CA (Italian Liver Cancer) database wasdivided in training (n = 3628, 70%) and validation (n = 1555, 30%)cohorts. A further cohort of 2651 HCC patients from Taiwan wasused for external validation. We defined the ITA.LI.CA Tumor Sta-tus (TS) using BCLC stages (0, A, B, C) but adding 3 intermediateB sub-stages: B1 = single > 5 cm or 2-3 nodules 3-5 cm; B2 = 2-3 nodules > 5 cm or > 3 nodules ≤ 5 cm; B3 = > 3 nodules > 5 cm orpresence of intrahepatic macrovascular invasion. TS was thenincluded in a multivariate survival model together with ChildPugh Score (CPS) and ECOG performance status (PST), and a novelITA.LI.CA prognostic staging was developed.

Results The ITA.LI.CA staging system was based on the follow-ing stages: 0n, An, B1n, B2n, B3n, Cn, where capital letters definedTS stages indicating potential for treatment as in BCLC (from rad-ical to palliative therapies), while n defined an additional score(0/1 = eligible for HCC therapy; 2/ > 2 = contraindication to therapy)based on CPS and PST indicating treatment feasibility. The concord-ance (c)-statistics for this model in training, internal and externalvalidation cohorts were respectively 0.72, 0.71, and 0.78, and theywere superior to that for BCLC (0.65, 0.64, and 0.73), CLIP (0.69,0.68, and 0.75), JIS (0.67, 0.67, and 0.70), MESIAH (0.69, 0.69, and0.77), and HKLC (0.68, 0,68, and 0.75).

Conclusion We developed a novel ITA.LI.CA staging system, easy
to apply in clinical practice, and with a strong prognostic impact intwo large western and eastern populations.

1 These authors equally contributed to this work.

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-12

IR-106B FOR HCC PATIENT MANAGEMENTFTER THERAPY

. Pascut 1,∗, R. Patti 2, N. Mezzina 3, C. Abazia 3,. Masutti 3, S.L. Crocè 2,3, C. Tiribelli 1,2,3

Fondazione Italiana Fegato, AREA Science Parkasovizza, Trieste, ItalyDepartment of Medicine, Surgery, and Healthciences, University of Trieste, Trieste, ItalyDepartment of General Surgery, Teaching Hospitalattinara, Trieste, Italy

Background: MicroRNAs (miRNAs) are regulatory noncodingNAs. Their aberrant expression is observed in many diseases,

ncluding Hepatocellular carcinoma (HCC). They are stable in bio-ogical fluids making miRNAs promising class of potential nonnvasive blood biomarkers for patients’ follow-up. mir-106b is fre-uently de-regulated in HCC with a role in cell proliferation andigration. Recent findings reported a circulating mir-106b altered

xpression in HCC.Aim: To investigate the role of miR-106b as peripheral blood

iomarker in HCC.Methods: 15 consecutive patients with early/intermediated

tage HCC were enrolled. Patients were treated according to theSSL/AASLD practice guidelines. All patients were staged at time 0,and 6 months after therapy with CT scan and/or MRI. The longest

ollow-up was 20 month. Stabilized Pax-gene tubes where usedo collect total blood before (T0) and one month (T1) after thereatment. Small RNA were purified and quantitatively analyzedy RT-qPCR. The relationships between their expression levelsnd clinicopathological parameters were further determined. Theaplan-Meier model was used to estimate disease-free survival

DFS).Results: A correlation between miRNA levels and treatment

esponse was observed (P < 0.001), 60% of the patients showed aomplete responseto treatment with a median mir-106b expres-ion of 2.4 at T1. The 40% showed a partial response or progressiveisease (median mir-106b expression = 0.78). Kaplan-Meier esti-ates and the log-rank test showed that high expression ofir-106b correlated with the longest DFS (P < 0.0038). Patients

n = 8) with higher miR-106b expression had a longer DFS timemedian = 15.5 months) as compared to patients (n = 7) with lowerxpression (median = 1 months). The expression of miRNA-106bas significantly correlated with the with BCLC staging A1, A2 and3 (P < 0.001).

Conclusions: Circulating mir-106b detection appears as a
romising non invasive marker to identify patients with longer DFS
n response to anticancer therapies.


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T-13

OLEUROPEIN REDUCES INFLAMMATORYMEDIATORS AND HEPATIC IMMUNE CELLSINFILTRATION IN A MOUSE MODEL OF NAFLD

M. Arciello 1,2,∗, B. Barbaro 1, A. Longo 1,R. Maggio 1, C. Viscomi 1, C. Balsano 1,3

1 Laboratory of Molecular Virology and Oncology,Francesco Balsano Foundation, Rome, Italy2 Department Internal Medicine and MedicalSpecialties, “Sapienza” University of Rome, Rome,Italy3 Institute of Molecular Biology and Pathology(IBPM) - CNR, Rome, Italy

Introduction: The metabolic syndrome (MeS) is a cluster ofmetabolic abnormalities like obesity, insulin-resistance and car-diovascular disease, with a growing worldwide prevalence thatreaches about 40% in population over 50 years of age. Nonalco-holic fatty liver disease (NAFLD) is considered a pathogenic factorof MeS as well as its hepatic manifestation. In some cases NAFLDmay progress from simple steatosis to end-stage liver diseases.Olives, main component of the Mediterranean diet, exerts bene-ficial effects on liver and heart, and reduces inflammation in MeSpatients, probably due its polyphenols such as oleuropein.

Aim: We aim to assess whether oleuropein supplementationmay counteract metabolism alterations and inflammation pro-duced by an excessive fat intake.

Methods: As model for NAFLD we used C57BL/6 mice fed witha high fat diet (HFD). After 8 weeks of HFD feeding, mice receiveda HFD supplemented with 3% oleuropein (OLE) for further 8 weeks(HFD + OLE).

Results: After preliminary evaluation of oleuropein efficacy inin vitro model of steatosis, we evaluated its effects in mouse modelof NAFLD. After 16 weeks, HFD-fed mice show elevated fat deposi-tion, increased body (BW), liver (LW) and heart (HW) weights andan increase of several circulating cytokines. HFD + OLE mice showreduced weight gain (BW - 25%, LW–50%, HW–70%) compared toHFD-fed mice, reduced liver damage and inflammatory infiltra-tion. Moreover, using the Bio-Plex multiplex biometric ELISA-basedimmunoassay in HFD + OLE we appreciated a significant reductionof various cytokines, including monocyte chemoattractant protein1 (MCP1) and chemokine (C-X-C motif) ligand 1 (CXCL1). Interest-ingly, MCP1 and CXCL1 are renowned players in the recruitment ofimmune cells and their increase is correlated to MeS.

Conclusion: These results suggest that oleuropein may preventNAFLD progression and MeS occurrence counteracting immune
cells infiltration in the liver, a key event in the progression of hepaticdamage.

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-14

NCIDENCE AND RISK FACTORS OFEPATOCELLULAR CARCINOMA IN UNTREATEDUBJECTS WITH CHRONIC HEPATITIS B VIRUSNFECTION: A SYSTEMATIC REVIEW AND

ETA-ANALYSIS

. Raffetti 1,∗, G. Fattovich 2,3, F. Donato 1

Unit of Hygiene, Epidemiology and Public Health,niversity of Brescia, Brescia, ItalyDepartment of Medicine, University of Verona,erona, ItalyClinical Unit of Gastroenterology, Aziendaspedaliera Universitaria Integrata Verona, Verona,

taly

Introduction and aims. The hepatocellular carcinoma (HCC) riskn chronic hepatitis B virus (HBV) infection may vary in differentlinical settings. We aimed to assess incidence rates, and factorselated with occurrence, of HCC in untreated subjects with HBVhronic infection.

Materials and Methods. We performed a systematic review andeta-analysis searching for published articles in Medline, Embase,

nd the Cochrane Library up to October 2014 as well as in refer-nce literature. We included longitudinal studies assessing HCCncidence in untreated HBV infected patients. The HCC incidenceates (IRs) estimates and their 95% confidence intervals (95% CIs)ere extracted by each study and pooled together in random effectsodels. Assessment of heterogeneity and meta-regression analy-

es were done.Results. 79 studies were included with a total of 363,838 par-

icipants and 3,472 HCC cases. In asymptomatic carries, the HCCRs were 0.04 (95%CI: 0.012-0.07), 0.19 (0.07-0.31) and 0.44 (0.23-.66) per 100 person-years in Europe, North America (around 70%siatic subjects) and East Asia, respectively. In inactive carriers andubjects with chronic hepatitis, the HCC IRs were 0.02 (0.0-0.06)nd 0.12 (0.03-0.27) in Europe, and 0.05 (0.03-0.06) and 0.49 (0.32-.66) in East Asia, respectively. In Child-Pugh A cirrhosis, IRs were.03 (1.30-2.77), 2.89 (1.23-4.55) and 3.62 (2.65-4.58) in Europe,orth America and East Asia, respectively. Meta-regression analy-

is showed a statistically significant IR increase with increasing ageevery 10 years: 1.09; 1.05-1.14), male percentage (1.02; 1.00-1.05),n North America (3.10; 1.00-9.98) and East Asia (2.24; 1.04-4.82)

ith respect to Europe, and in asymptomatic carriers (1.84; 0.92-.66), and patients with chronic hepatitis (7.84; 3.44-17.98) andompensated cirrhosis (18.92; 9.93-35.71) with respect to inactivearriers.

Conclusions. The risk of developing HCC in untreated subjects
ith HBV chronic infection is strongly influenced by age, gender,acro-area of origin, and HBV liver disease status.

sease 47S (2015) e19–e42 e25

T-15

ADHERENCE TO AISF RECOMMENDATIONS FORINTEGRATED MANAGEMENT OFHEPATOCELLULAR CARCINOMA IN AREAL–WORLD CLINICAL PRACTICE

G. Piai 1, V. Messina 2, G. Valente 1, L. Rinaldi 1,G. Moggio 3

1 UOSD Fisiologia Epatica con Servizio di Assistenzaai Trapiantandi e Trapiantati Epatici, AORNSant’Anna e San Sebastiano, Caserta, Italy2 UOS Ottimizzazione e Monitoraggio delle EpatitiCroniche Virali UOC Malattie Infettive, AORNSant’Anna e San Sebastiano, Caserta, Italy3 UOSD Radiologia Interventistica, AORN Sant’Anna eSan Sebastiano, Caserta, Italy

Introduction and Aim: AISF recommendations (AISF-R) forHCC management are based on the assumption of a multidisci-plinary approach in a network with availability of all diagnosticand therapeutic resources. In our real world, however, a specializedhepatic surgery unit is missing. Hepatologists, radiologists, oncolo-gists were then all invited to join a multidisciplinary HCC-team thatwe connected to many centres of liver surgery throughout Italy.

Aim of study was to evaluate the efficiency of this organizedapplication of AISF-R, considered as a methodological effort toimprove quality of assistance and to reduce the high health mobilityfrom our geographic area.

Methods: One hundred twenty-six HCC patients, stratifiedaccording to BCLC system, were discussed by HCC-team (January2012 - September 2014). Each formal clinical record was then inde-pendently revised, expressing a “theoretical” indication accordingto AISF-R and comparing it to the really applied choice.

Results: In 77 cases surgery appeared obviously not indicatedor contraindicated, while liver surgery advice was requested in 49cases (39%): subdivided in BCLC A1-A3/A4/B/C/D groups, rates were100%, 2%, 37%, 26%, 17%, respectively.

Overall, 69% of patients were treated according to AISF-R. Adher-ence was 11/12 (92%) early HCC theoretically eligible for resection,10/21(48%) liver transplantation, and 21/28 (75%) percutaneoustreatments (PT). TACE was performed in 12/61 (20%) early HCC,in 3/31 (10%) advanced HCC, and excluded in 14/34 (41%) inter-mediate HCC. Resection was performed in 11/12 (92%) early HCCeligible for PT and in 1/33 (3%) advanced HCC.

Conclusions: The method of a multidisciplinary HCC-team 1)seems to overcome some limitations due to local unavailability ofliver surgery, as shown by high rates of adherence to qualified rec-ommendations, with well-founded reasons for discrepancies and
for referring to liver surgery centres; 2) provides the advantages ofa collegial choice and often reduces useless/costly patient mobility.

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-16

OST-TRANSLATIONAL REGULATION OFOLYCYSTIN 2 (PC2)EXPRESSION AS A NOVELECHANISM OF CHOLANGIOCYTE REACTION TO

ILIARY DAMAGE AND REPAIR

. Spirli 1,∗, A. Villani 1, C.M. Morell 3, L. Fabris 2,

. Fiorotto 1, M. Strazzabosco 1,3

Section of Digestive Diseases, Yale University, Newaven, Connecticut, USADepartment of Molecular Medicine, University ofadua, Padua, ItalyDepartment of Surgery and Interdisciplinaryedicine, University of Milan-Bicocca, Milan, Italy

Introduction: In polycystic liver diseases, defective PC2 leadso elevated production of cAMP, PKA-dependent activation ofhe ERK1/2 pathway, HIF1�-mediated VEGF production andtimulation of cyst growthand progression. Activation of theRK/HIF1�/VEGF pathway in cholangiocytes is fundamental dur-ng repair from biliary damage. In this study we hypothesized thatC-2 levels can be modulated during biliary damage/repair.

Results: PC2 protein expression was significantly reduced inivers from mice undergoing cholangiocyte damage (Mdr2-/--KO,DL, treatment with dehydrocollidine–DDC). However, PC-2-genexpression was not reduced,suggesting that the decrease in PC2as due to increased degradation. To understand if factors involved

n biliary damage influence PC2 protein expression, mouse cholan-iocytes were treated with pro-inflammatory cytokines, nitricxide (NO) donors and ER stressors. Expression of PC2 proteinas again significantly reduced, but not its gene expression.oteworthy, downregulation of PC-2 leads to increased ERK1/2hosphorylation, HIF1� transcription activity and secretion ofEGF. Expression of Herp and NEK, proteins that promote PC2egradation via the proteasome complex was also increased.re-treatment with proteasome inhibitor restored the expressionf PC2 in cells treated with cytokines but not in cells treatedith NO donors or with ER stressors. In the latter conditions,

C2 degradation was inhibited by inhibition of PI3 K, that blocksutophagosome and by blockade of lysosomes, suggesting a role forutophagy. Finally, treatment of DDC-treated mice with the protea-ome inhibitor bortezomib, significantly reduced the extent of theiver damage and restored PC2 expression.

Conclusion: PC2 is modulated post-translationally by pro-nflammatory cytokines, ER-stressors and NO-donors ands reduced in mice with biliary damage. Down regulationf PC2 protein expression in cholangiocytes increases theRK1/2/HIF1�/VEGFsecretion, thereby playing a pivotal rolen the regulation of cholangiocyte response to biliary dam-
ge.Treatments able to restore PC2 expression may represent aew therapeutic approach in biliary diseases.

sease 47S (2015) e19–e42

T-17

ANTIVIRAL THERAPY WITH SOFOSBUVIR PLUSRIBAVIRIN FOR THE TREATMENT OF SEVEREHCV RECURRENCE AFTER LIVERTRANSPLANTATION: PRELIMINARY DATA FROMA SINGLE-CENTRE EXPERIENCE

D. Arese 1,∗, S. Martini 1, S. Strona 1, M. Sacco 1,D. Cocchis 2, S. Mirabella 2, G. Rizza 2, F. Tandoi 2,R. Romagnoli 2, A. Ottobrelli 1,M.R. Torrani-Cerenzia 1, F. Balzola 1, M. Salizzoni 2,M. Rizzetto 1

1 Gastrohepatology Unit, A.O.U. Città della Salute edella Scienza di Torino, Torino, Italy2 Liver Transplant Centre, General Surgery Unit,A.O.U. Città della Salute e della Scienza di Torino,Torino, Italy

Background and aims: Recurrent hepatitis occurs in the majorityof HCV viremic recipients at liver transplantation (LT). We evalu-ated safety and efficacy of antiviral therapy (Tx) with sofosbuvir(SOF) plus ribavirin (RBV) in a population of patients (pts) havingsevere recurrent hepatitis after LT.

Methods: Among HCV viremic pts transplanted in our Centresince 2000, we enrolled to date 107 pts affected by HCVRNA-positive recurrent hepatitis with fibrosis score F3 (21 pts) or F4 (86pts) according to Metavir. 78% male, mean age 60.5 years, meanBMI 24.7; 49% receiving cyclosporine, 43% mycophenolate, 38%tacrolimus and 3% m-TOR inhibitors. GT1 75% (GT1a 13%), GT2 5%,GT3 15%, GT4 5%; IL28CC 24%; 65% experienced. According to com-passionate use, pts receive SOF 400 mg/die plus RBV (weight-based)for 24 weeks. Median time from LT to Tx 4.96 years (0.25-14.25);mean baseline HCVRNA 6.62 log10 IU/mL (3.4-7.9 log10 IU/mL);mean GFR 76 mL/min.

Results: Until now, 31/107 pts completed 12 weeks of Tx. After1 week of Tx, median decrease in HCVRNA was 3.61 log10 IU/mL;50/98 (51%) at week 4, 76/77 (99%) week 8, 31/31 week 12, 9/9week 16, 2/2 week 24 on Tx were HCVRNA negative. Of the 2pts who completed the planned therapy and reached 4 weekspost-Tx, one relapsed; he is IL28TT, GT1a and the only one whotested HCVRNA < 15 IU/mL at w8 on Tx. The most common adverseevents were fatigue, headache and nausea. During Tx, 1 patient hadvariceal bleeding (day 78) and 1 died due to multiorgan failure (day86); 25% received blood transfusion/epoetin. Minimal immunosup-pression dose adjustments were required on Tx and no rejectionswere recorded.

Conclusions: In pts with severe HCV recurrence post-LT, antivi-


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-18

SSESSMENT OF FREE LIGHT CHAINS IN HCVOSITIVE PATIENTS WITH MIXEDRYOGLOBULINEMIA VASCULITIS UNDERGOINGITUXIMAB TREATMENT

. Basile 1,∗, L. Gragnani 2, E. Torti 1, A. Piluso 2,. Gulli 1, T. Urraro 2, M.T. Dell’Abate 1, C. Stasi 2,. Monti 2, G.L. Rapaccini 3, A.L. Zignego 2

Department of Laboratory Medicine of the Catholicniversity of the Sacred Heart, Rome, ItalyDepartment of Experimental and Clinical Medicine,niversity of Florence, Florence, ItalyInstitute of Internal Medicine, Catholic Universityf the Sacred Heart, Rome, Italy

Introduction: Mixed Cryoglobulinemia (MC) is an HCV-relatedymphoproliferative disorder, secondary to a systemic vasculitis ofmall vessels. Treatment with anti-CD20 monoclonal antibodiesituximab (RTX) is proved to be very useful. Free light chain (FLC)/� ratio and FLC patterns were associated with MC and/or B-nonodgkin’s lymphoma.

Aims: The aim of this study was to evaluate changes in serumree light chains (FLC) in HCV positive patients with related Mixedryoglobulinemia (MC) undergoing anti-CD20 monoclonal anti-ody Rituximab (RTX) therapy. Furthermore, we attempted toorrelate FLC values with therapy response.

Patients and Methods: We retrospectively enrolled 46 patientsith HCV infection (26 females, 20 males), including 10 patientsithout signs/symptoms of MC-related vasculitis, 36 with MC-

asculitis. Clinical and biological data were recorded at baselinend six months after RTX treatment. Nephelometric measurementf serum FLCs was performed using a serum FLC assay.

Results: The mean serum FLC-k level was significantly higher inC patients, compared to HCV patients without MC and to blood

onors (p = 0.05 and p < 0.0001, respectively); the mean serum FLC-atio was significantly higher in MC patients, compared to HCVatients without MC and to blood donors (p = 0.0023 and p = 0.0008,espectively). An abnormal FLC-ratio at baseline correlated withresence of cryoglobulins, C4 consumption, higher RF level andigher vasculitis rate (p < 0.005 for each parameter).

In order to evaluate the predictive value of FLC patterns, MCatients were divided into two groups according to RTX therapyutcome (responders and no/partial responders).

Abnormal baseline FLC-ratio was significantly associated witho/partial response (OR 4.86–95% C.I. 0.89-28.72, p = 0.0314).

Conclusions: RTX-treatment in HCV-related MC induces aeduction of FLC-k and RF levels. Moreover, pre-treatment FLC-atio, which can be easily assessed by a routine test, may be useful
o predict response to this expensive treatment for HCV-related MCatients ineligible to IFN-based therapy.

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T-19

PNPLA3 RS738409 I748 M IS ASSOCIATED WITHSTEATOHEPATITIS IN NON OBESE SUBJECTSWITH HEPATITIS C

S. Petta 1,∗, E. Vanni 2, E. Bugianesi 2, C. Rosso 2,D. Cabibi 3, C. Cammà 1, V. Di Marco 1, M. Eslam 4,S. Grimaudo 1, F.S. Macaluso 1, D. McLeod 5,R.M. Pipitone 1, M.L. Abate 2, A. Smedile 2,J. George 4, A. Craxì 1

1 Section of Gastroenterology, Di.Bi.M.I.S., Universityof Palermo, Palermo, Italy2 Division of Gastro-Hepatology, Department ofMedical Sciences, San Giovanni Battista Hospital,University of Turin, Turin, Italy3 Cattedra di Anatomia Patologica, University ofPalermo, Palermo, Italy4 Storr Liver Unit5 Institute of Clinical Pathology and MedicalResearch, Westmead Millennium Institute andWestmead Hospital, University of Sydney, Sydney,Australia

Background and Aims: The PNPLA3/Adiponutrin rs738409 C/Gsingle nucleotide polymorphism is associated with the severity ofsteatosis, steatohepatitis and fibrosis in patients with non-alcoholicfatty liver disease, as well as with the severity of steatosis and fibro-sis in patients with chronic hepatitis C (CHC). We tested in genotype1 (G1) - CHC patients the putative association between the PNPLA3variant and histological features of steatohepatitis, as well as theirimpact on the severity of fibrosis.

Methods: 434 consecutively biopsied Caucasian G1-CHCpatients were genotyped for PNPLA3 rs738409. Histological fea-tures of steatohepatitis in CHC were assessed using the Bedossaclassification. Hepatic expression of PNPLA3 mRNA was evaluatedin 63 patients.

Results: The prevalence of steatohepatitis increased from 16.5%in patients with PNPLA3 CC, to 23.2% in CG and 29.2% in GG geno-type (p = 0.02). By multiple logistic regression, PNPLA3 genotype(OR 1.54, 95%CI 1.03-2.30, p = 0.03) together with age (OR 1.03,95%CI 1.00-1.05, p = 0.02), BMI ≥ 30 (OR 2.06, 95%CI 1.04-4.10,p = 0.03) and HOMA (OR 1.18, 95%CI 1.04-1.32, p = 0.006) wereindependently linked to steatohepatitis. When stratifying for obe-sity, PNPLA3 was associated with NASH in non obese patientsonly (12.0% in CC vs 18.3% in CG vs 27.3% in GG, p = 0.01), includ-ing after correction for metabolic confounders (OR 2.06, 95%CI1.26-3.36, p = 0.004). We confirmed the independent associationof rs738409 with the severity of steatosis (OR 1.71, 95%CI 1.20-2.45, p = 0.003), and, indirectly by steatohepatitis promotion (OR2.05, 95%CI 1.05-4.02, p = 0.003), with severe fibrosis. Higher liverPNPLA3 mRNA was associated both with the severity of steatosis(adjusted p = 0.03) and steatohepatitis after adjusting for gender,age, BMI and HOMA (p = 0.002).

Conclusions: In G1-CHC patients, the PNPLA3 G variant is asso-
ciated with a higher risk of steatosis severity and steatohepatitis inchronic hepatitis C, particularly among non obese subjects.

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-20

LA DPB1 RS9277535 POLYMORPHISMTRONGLY PREDICTS HBSAG CLEARANCE INNTERFERON TREATED GENOTYPE DBEAG-NEGATIVE PATIENTS WITH CHRONICEPATITIS B

. Lampertico 1,∗, E. Galmozzi 1, F. Facchetti 1,

. Cheroni 2, F. Invernizzi 1, V. Valveri 2,

. Soffredini 1, M. Viganò 3, S. Abrignani 2,

. De Francesco 2, M. Colombo 1

Division of Gastroenterology and Hepatology,ondazione IRCCS Cà Granda Ospedale Maggioreoliclinico, University of Milan, Milan, ItalyINGM Istituto Nazionale Genetica Molecolareilan, ItalyDivision of Hepatology, Ospedale San Giuseppe,niversity of Milan, Milan, Italy

Introduction and Aim: HLA DP polymorphisms have recentlyeen associated to spontaneous hepatitis B virus (HBV) clearance insians patients. Whether these SNPs influence IFN-induced HBsAgesponse in difficult to cure HBeAg-negative genotype D chronicepatitis B Caucasian patients, is currently unknown.

Materials and Methods, Results: 126 such patients with com-ensated disease (46 years, 82% males, 90% genotype D, HBV DNA.2 log cp/ml, ALT 132 IU/L, 40% with cirrhosis) were followedor a median of 11 (1-23) years after 22 (6-48) months of eithertandard or pegylated (Peg)IFN alfa treatment. The primary end-oint was HBsAg clearance. HLA DPA1 rs3077, HLA DPB1 rs9277535nd rs9277534, and IL28B rs12979860 genotypes were assessedy using TaqMan SNP Genotyping Assays. Twenty-eight (22%)atients ultimately cleared serum HBsAg with a 15-year cumula-ive probability of 30%, with a preference for patients with baselineower HBV DNA levels, higher ALT levels, IL28B rs12979860 CC,LA DPB1 rs9277534 AG/GG and HLA DPB1 rs9277535 AG/GG.t multivariate analysis, HLA DPB1 rs9277535 AG/GG genotypeas the strongest independent predictor of HBsAg seroclearance

HR: 6.88; 95%CI 2.82-16-7, p < 0.000) along with baseline ALT lev-ls (HR 1.00, 95%CI 1.00-1.00, p = 0.001), baseline HBV DNA levelsHR 0.54, 95%CI 0.37-0.79, p = 0.002), and IL28B rs12979860 CCenotype (HR 2.2, 95%CI 1.02-5.0, p = 0.044). The 15-year cumu-ative rates of HBsAg clearance were 62% in carriers of the HLAPB1 rs9277535 AG/GG genotype compared to 13% in carriers of

he AA genotype (p < 0.0000). Conclusions: HLA DPB1 rs9277535olymorphism strongly predicts IFN-induced HBsAg clearance inBeAg-negative patients chronically infected by genotype D HBV.
his genetic signature may help to select patients to IFN basedegimen.

sease 47S (2015) e19–e42

T-21

ADD-ON PEGINTERFERON ALFA-2ASIGNIFICANTLY REDUCES HBSAG LEVELS INCHRONIC HEPATITIS B, HBEAG-NEGATIVE,GENOTYPE D PATIENTS FULLY SUPPRESSED ONNUCLEOT(S)IDE ANALOGUES TREATMENT:HERMES STUDY INTERIM ANALYSIS

P. Lampertico 1,∗, M.R. Brunetto 2, A. Craxì 3,G.B. Gaeta 4, M. Rizzetto 5, G. Palmieri 6,M. Colombo 1

1 Division of Gastroenterology and Hepatology,Ospedale Maggiore Policlinico, University of Milan,Milan, Italy2 Liver Unit, Reference Centre for Chronic LiverDesease and HCC of the Tuscany Region, UniversityHospital of Pisa, Pisa, Italy3 Gastroenterology and Hepatology, Di.Bi.M.I.S.,University of Palermo, Palermo, Italy4 Department of Internal Medicine, Section ofInfectious Desease, Second University of Naples,Naples, Italy5 Department of Gastroenterology, University ofTurin, Turin, Italy6 Roche S.p.A., Monza, Italy

Backgound: Since nucleos(t)ide Analogues (NA) suppress viralreplication in HBeAg-negative CHB without inducing significantlyHBsAg loss, several studies are investigating the impact to HBsAglevels of Peginterferon alfa-2a (PEG-IFN) addition to NA treatedpatients. Here we present the week 24 analysis of an ongoing trialevaluating PEG-IFN add-on in NA treated CHB, HBeAg-negative,genotype D, Caucasians patients (HERMES study). Methods: In thisphase IIb, open label, single arm, multicenter Italian study, patientswho were receiving NA monotherapy, with HBV-DNA persistentlybelow 20 IU/ml for at least 12 months and HBsAg > 100 IU/ml, ini-tiated add-on treatment with Peginterferon alfa-2a 180 �g sc onceweekly for 48 weeks. Patients without any HBsAg decrease at week24 or those who will complete the add-on treatment period (week48) are going to discontinue PEG-IFN and be followed-up for addi-tional 48-weeks. Serum HBsAg decline at the end of treatmentwith PEG-IFN plus NA combination (week 48) is the primary studyendpoint. Results: 70 Caucasians who had completed 24 weeks ofcombination treatment were included in this analysis. Median agewas 50.5 (29-64) and 81% were males. At week 24 of combina-tion treatment, 27.1% decreased serum HBsAg≥50% from baseline,while 15.7% discontinued study due to lack of response. MedianqHBsAg significantly decreased from 1160 IU/ml at baseline to743 IU/ml at week 24 (p < 0.0001). The proportion of patients withHBsAg < 500 or < 1000 IU/ml increased from baseline (23% and 47%respectively) up to week 24 (39% and 59%, respectively). Fifty-sevenpatients (81.4%) experienced adverse events (AEs), 4.3% and 5.7% ofthe patients discontinued or temporarily interrupted PEG-IFN dueto AEs respectively, while in 8.6% of patients PEG-IFN dosage wasadjusted due to AEs. Conclusion: In HBeAg-negative, CHB, genotype
D patients treated with nucleot(s)ide analogues, add-on treatmentwith PEG-IFN results in significant reduction of serum HBsAg levels.

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-22

OLE OF KILLER CELL IMMUNOGLOBULIN-LIKEECEPTORS AND THEIR HLA CLASS I LIGANDS INUTOIMMUNE HEPATITIS

. Littera 1, C. Carcassi 1, L. Secci 2, S. Lai 1,. Cappai 1, R. Porcella 1, F. Alba 1, R. Maddi 1,. Serra 1, S. Cappellini 2, C. Salustro 2, S. Onali 2,

. Balestrieri 2, G. Serra 2, M. Conti 2, T. Zolfino 3,

. Scioscia 2, L. Barca 2, L. Chessa 2

Medical Genetics, Department of Medical SciencesM.Aresu”, University of Cagliari, ItalyCenter for the Study of Liver Diseases, Departmentf Medical Sciences “M. Aresu”, University of Cagliari,talyS.C. Gastroenterologia, AOB, Cagliari, Italy

Introduction: It is well known that CD4, and CD8 T lymphocytesre critical protagonists of the immunopathogenetic mechanismsf autoimmune hepatitis (AIH). However, so far, data on thenvolvement and role of natural killer (NK) cells are scarce.

Aim: Killer cell immunoglobulin receptors (KIRs) represent theajor family of cell surface receptors that inhibit and activate NK

ells. The elevated expression of NK cells in liver tissue prompteds to evaluate the impact of NK cells and KIRs on age of onset andvolution of the disease.

Materials and Methods: We retrospectively analyzed a totalf 114 Sardinian patients diagnosed with type I AIH. Metabolicnd genetic causes of hepatopathy were excluded in all patients.atients and controls were typed at high resolution (4 digits) forhe alleles at the HLA-A, -B, -C and DR loci and at 15 KIR geneoci. Patients were divided into 2 groups according to homozygosityor KIR haplotype A (KIR genotype AA), heterozygosity or homozy-osity for KIR haplotype B (KIR genotype Bx).The immunogeneticharacteristics of the AIH patients were compared with those of21 healthy individuals from the Sardinian bone marrow donoregistry.

Results: The activating KIR gene KIR2DS1 had a significantlyigher frequency in AIH patients compared to controls [57% vs3%; HR (95% CI) = 1.7 (1.0-2.7); P = 0.03]. Also the presence ofIR2DL1 + /KIR2DS1 + /HLA-C2 + was higher in AIH patients [47.3%s 35.7%; HR (95% CI) = 1.6 (1.0-2.6); P = 0.04].

Conclusions: Based on our findings, it can be hypothesized thatK cells are involved in the pathogenetic mechanisms of type I AIHnd more specifically, that the activating KIR gene KIR2DS1 is asso-
iated with the development of the disorder. The role of KIR2DS1s a marker for AIH warrants further investigation.

sease 47S (2015) e19–e42 e29

T-23

SLOW ACHIEVEMENT OF HCV-RNAUNDETECTABILITY IN CIRRHOTIC PATIENTSTREATED WITH SOFOSBUVIR + RIBAVIRIN:POSSIBLE CLINICAL IMPLICATIONS IN THE LIVERTRANSPLANT LIST MANAGEMENT

I. Lenci 1, V. Cento 2, M. Rendina 3, M.F. Donato 4,M. Milana 1, D. Sforza 5, M. Manuelli 5, M. Aragri 2,V.C. Di Maio 2, A. Abedrabbo 1, A. Castellaneta 3,F. Malinverno 4, S. Monico 4, M.L. Ponti 6,R. Canu 6, R. Ganga 6, R. Alfieri 7, L. Milanesi 7,A. Di Leo 3, G. Tisone 5, C.F. Perno 2,8,F. Ceccherini-Silberstein 2, M. Colombo 4,M. Angelico 1

1 Hepatology Unit, Policlinico Tor Vergata, TorVergata University, Rome2 Virology Chair, Tor Vergata University, Rome3 Gastroenterology and Digestive Endoscopy,University Hospital, Policlinico Bari4 Gastroenterology and Liver Transplant Units, CàGranda Maggiore Hospital Policlinico and Universityof Milan, Milan, Italy5 Liver Transplant Centre, Policlinico Tor Vergata, TorVergata University, Rome6 Grastroenterology and Hepatology Units, BrotzuHospital, Cagliari, Italy7 Institute for Biomedical Technologies-CNR, Segrate,(Milano)8 Virology Unit, Policlinico Tor Vergata, Tor VergataUniversity, Rome

Sofosbuvir (SOF) treatment ± ribavirin (RBV) prior to LT has thepotential to change the HCV recurrence after Liver Transplantation(LT). This approach has been reported to avoid graft reinfectionin compensated patients with hepatocellular carcinoma (HCC),but only among those who reached and maintained undetectableHCV-RNA (TND) before LT. Since the time to obtain this result indecompensated cirrhotics is unknown, we sought to investigatethe early HCV-RNA decay in this setting.

Sixteen decompensated patients (M/F 12/4, median age 55.3,CPT score ≥ B7), infected by HCV genotype 1a, 1b, 3 and 4 (2-8-4-2), 4 of whom with HCC, were treated with SOF 400 mg/dayand RBV (200-1000 mg/day), except 2 who received SOF alone, fora median(IQR) of 12(11-16) weeks awaiting LT. HCV-RNA levelswere measured weekly and safety and clinical parameters wereanalyzed.

No serious adverse events were reported. The median(IQR)RBV dose was 600(400-800). Despite 11/16 patients had lowbaseline viremia (<600.000 IU/ml), HCV-RNA decay in the first 4weeks of treatment was suboptimal (median[IQR] = -3.7[-4.3;-3.3]LogIU/ml). Only 3/16 (18.7%) patients reached TND HCV-RNA atweek 4 (rapid virological response, RVR), and 5/10 (50%) evalu-able patients were still viremic at week-12. Median(IQR) MELD
decreased from 15(13-16) at baseline to 13(12-15) at week 4,when 14/16 (87.5%) patients returned in a compensated stage. Onepatient underwent LT after 6 weeks of treatment, while HCV-RNAwas still positive (26 UI/mL). SOF was interrupted only during the

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30 Abstracts / Digestive and Liver Disease 47S (2015) e19–e42

rst 4 perioperative days, obtaining TND HCV-RNA at week 10 ofherapy.

The kinetics of HCV-RNA decay in decompensated cirrhoticatients are slower compared to those obtained in non cirrhoticatients. As a practical implication, pre-LT treatment may need toe longer and/or based on more effective antiviral strategies. Alter-atively, MELD-based prioritization to LT should be reassessed in

ight of the results of antiviral therapy.


-24

HRONIC HEPATITIS B TREATMENTNDIVIDUALIZATION BY MEANS OF SERUMBSAG AND MIR-B-INDEX KINETICS

. Cavallone 1, F. Oliveri 1, P. Colombatto 1,. Coco 1, P. Ciccorossi 1, V. Romagnoli 1,. Cherubini 1, F. Moriconi 1, F. Bonino 2,.R. Brunetto 1

Hepatology Unit, Reference Center of the Tuscanyegion for Chronic Liver Disease and Cancer,niversity Hospital of Pisa, ItalyDigestive and Liver Disease, General Medicine IInit, University Hospital of Pisa, Italy

Background and Aims Non-viral biomarkers of sustainedontrol of HBV infection are an unmet need for treatment indi-idualization in Chronic-Hepatitis-B (CHB). We reported thatnactive-Carriers and patients with sustained-virologic-responseSVR) to Peg-IFN share a common serum miRNA signature (MiR-B-ndex, MBI). We studied the kinetics of MBI and HBsAg during/aftereg-IFN.

Patients and Methods 46 CHB patients (11 HBeAg-pos, 29ales, median-age 48.6y, 22.4-64.1y; genotypes: A 5, D 39, F 2)ere treated with Peg-IFN2a 180 �g/w (median 13.2months, 8-

4 m): 20 SVR; 17 relapsers (REL) and 9 non-responders (NR).he quantitative analysis of miR-122-5p, miR-99a-5p, miR-192-5p,iR-126-3p, miR-335-5p and miR-320a was performed as pre-

iously reported. HBsAg and MBI were tested at baseline (BL),4-48w, end-of-therapy (EOT), 24w-post-treatment (PTFU).

Results MBI > -1.7 were found: at BL in 5 HBeAg-neg pts (4 SVR,NR); at EOT in 14/19 (73.7%) SVR (4 HBeAg-pos, 10 HBeAg-neg)

nd at PTFU in all SVR, but in 1 NR and 1 REL. Kinetics of HBsAgnd MBI in HBeAg-neg pts are reported in the table; MBI≥-3.0nd HBsAg < 1000 IU/mL at EOT had: 100-93.3% sensitivity, 94.7-5.0% specificity, 93.3-82.4% PPV, 100-94.4% NPV, 97-88.6% DA in

dentification of SVR (AUROC 0.988 and 0.951 respectively).Conclusions Overall HBsAg and MBI show good performances

n SVR prediction. In HBeAg-neg-CHB MBI predicts SVR since ther-py beginning and identifies all SVR at EOT qualifying as the bestiomarker for individual therapy monitoring.

HBsAg (Log10IU/mL) MiR-B-IndexBL T24w T48w EOT PTFU P** BL T24w T48w EOT PTFU P**

SVR(15) 3.45 2.81 2.69 2.15 1.33 .002 −4.49 −1.65 1.16 1.46 4.60 < .001REL(15) 3.76 3.63 3.50 3.44 3.34 .008 −9.81 −8.31 −8.80 −7.22 −4.90 .034NR(5) 3.70 3.60 3.43 3.58 3.10 .607 −7.26 −8.29 −7.20 −8.82 −5.92 .335P* .042 .001 < .001 < .001 < .001 .001 < .001 < .001 < .001 < .001

T-25

HBX–DLEU2 LNCRNA COMPLEX AFFECTSTRANSCRIPTION OF NEW TARGET PROMOTERS

F. Guerrieri 1,2, L. Chiodo 1, S. Jeddari 2,D. D’Andrea 3, A. Tramontano 3, G. Ruocco 1,M. Levrero 1,2

1 CLNS@SAPIENZA, Istituto Italiano di Tecnologia(IIT), Rome, Italy2 Department of Internal Medicine, SapienzaUniversity, Rome, Italy3 Biocomputing Lab, Department of Physics, SapienzaUniversity, Rome, Italy

Background: A ChIPSeq analysis of HBx genome wide recruit-ment identified 39 long non coding RNAs (lncRNAs), includingDLEU2, as direct HBx transcriptional targets in HBV replicatingcells. DLEU2 lncRNA overlaps the first exon of the TRIM13 gene andthe pri-Mir of the miR-15a-16.1 cluster. Up-regulation of specificDLEU2 splicing variants correlates with tumors.

Objectives: Aim of this study was to investigate HBx role inDLEU2-TRIM13-mir15.mir16 regulation.

Methods: Anti-HBx ChIP were performed in HepG2 cells repli-cating wt and HBx-mt HBV. HBx ChIPed DNA and lncRNA-geneexpression levels were analysed by TaqMan RT-PCR using specificprimers. Specific LNATM longRNA GapmeRs (Exiqon) were used forhighly efficient inhibition of DLEU2 lncRNA function. The I-Tasser,RNAfold and 3dRNA softwares were used for prediction of HBx pro-tein structureand RNA secondary and tertiary structures.

Results: HBx binding to the DLEU2 promoter leads to increasedH4 acetylation, a different DLEU2 splicing profile, down-regulationof miR-15a-16.1 cluster and up-regulation of the antisenseautophagic gene TRIM13, in the absence of HBx binding to theTRIM13 promoter. GapmeR-mediated selective degradation ofDLEU2 RNA results in reduced TRIM13 promoter H4 acetylationand a ∼50% reduction of TRIM13 expression in HBV replicatingcells. These results directly link DLEU2 RNA species with TRIM13transcriptional regulation in the presence of HBx. The interactionbetween HBx and DLEU2 was confirmed by RIP (RNA ImmunePrecipitation) experiments. By coupling ab initio modeling of HBxprotein and DLEU RNA tertiary structures and we constructed adocking model of DLEU2-HBV complex. Finally, we found thatDLEU2 inactivation has a profound impact on pgRNA transcrip-tion, suggesting a functional relevance of DLEU2-HBx interactionfor HBV replication.

Conclusion: HBx targets the DLEU2 promoter leading to a dif-ferent DLEU2 splicing profile and its binding to DLEU2 RNA speciesaffects cellular genes transcription and HBV replication.


Mann-Whitney (SVR vs REL + NR); **ANOVA (BL vs EOT).


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-26

MPULSIONAL, POINT AND BIDIMENSIONALHARE WAVE ELASTOMETRY FOR PORTALYPERTENSION: SAME STIFFNESS THRESHOLD?

. Stefanescu, G. Allegretti, C. Serra, G. Marasco,

. Gamal, F. Conti, A. Colecchia, D. Festi,. Andreone, L. Bolondi, F. Piscaglia

Department of Medical and Surgical Sciences,.Orsola-Malpighi Hospital, Alma Mater Studiorum -niversity of Bologna, Bologna, Italy

BACKGROUND: Liver stiffness (LS) is used to investigate theresence of portal hypertension (PH). LS measured with Transientlastography (TE) correlates with HVPG, and 21.1 kPa is proposeds cut-off to predict clinically significant PH (CSPH). New elasto-raphic techniques may overcome some limitations of TE, but theiriagnostic accuracy remain to be defined. We aimed to comparehe performance of TE, point shear wave elastography (ElastPQ)nd 2D-Shear Wave Elastography (2D SWE) to predict the grade ofH in a cohort of cirrhotic patients.

METHODS: 65 consecutive patients (63.1%M, 60.2y) withhronic liver disease submitted to HVPG measurement and ateast one elastographic technique: TE and/or ElastPQ (Philips IU22)nd/or 2D SWE (Aixplorer by Supersonic Imagine) blindly one fromach-other, were enrolled between December 2013 and November014.

RESULTS: 44/65 patients (67.7%) had CSPH (HVPG > 10 mmHg).0 patients (61.5%) underwent TE, 31(47.7%) ElastPQ, 25(38.5%) 2D-WE measuring liver stiffness.

HVPG was well correlated with either TE (r = 0.695;p < 0.0001),lastPQ (r = 0.684;p = 0.001), or 2D-SWE (r = 0.762;p < 0.0001).

The overall concordance between elastographic techniquestested in 51 patients who underwent either 2 or 3 methods) wasnly moderate (ICC = 0.409). TE was concordant with both otherechniques (ICC = 0.772 for ElastPQ, ICC = 0.754 for 2D-SWE), whilelastPQ and 2D-SWE were less concordant (ICC = 0.488).

No significant difference was detected among AUROCs for pre-icting CSPH (0.872 for TE, 0.962 for ElastPQ and 0.860 for 2D-SWE,espectively).

For the 21.1 kPa cutoff, the accuracy of each technique to cor-ectly classify CSPH was: 85%(34/40) for TE, 55%(17/31) for ElastPQnd 76%(19/25) for 2D-SWE, respectively.

CONCLUSION: Our findings showed that liver stiffness, assessedy new elastographic techniques (ElastPQ and 2D-SWE), haveimilar performance to predict CSPH like TE. However the bestiagnostic threshold for this diagnosis differs significantly among
ifferent elastography technologies and specific diagnostic accu-acy studies appear warranted for each type of equipment.

sease 47S (2015) e19–e42 e31

T-27

SERPINB3 AND YAP INTERPLAY INCREASES MYCONCOGENIC ACTIVITY

C. Turato 1, S. Cannito 2, D. Simonato 1,G. Villano 1,2,3, L. Terrin 1, S. Quarta 1, A. Biasiolo 1,M. Ruvoletto 1, S. Fasolato 3, G. Zanus 3, U. Cillo 3,A. Gatta 1, M. Parola 2, P. Pontisso 1

1 Department of. Medicine, University of Padua,Padua, Italy2 Department of. Clinical and Biological Sciences,University of Turin, Turin, Italy3 Department of Surgical, Oncological andGastroenterological Sciences, University of Padua,Padua, Italy

Background SerpinB3 has been recently described as an earlymarker of liver carcinogenesis. Myc is one of the most importantoncogenes in human cancer. Somatic amplification and overex-pression of Myc often correlate with more advanced and aggressivetumour forms. Yes-associated protein (Yap), the main effector ofthe Hyppo pathway, is a central regulator of proliferation andself-renewal of normal and cancer stem cells. This molecule,found up-regulated in hepatocellular carcinoma (HCC), has beendescribed also to increase Myc expression.

Aim The present study has been designed in order to investi-gate whether SerpinB3 may functionally modulate Myc in differentexperimental models and in human HCC specimens.

Materials and Methods Expression of Myc, Yap and its targetgenes was evaluated in relation to SerpinB3 expression in HCCspecimens, in C57BL/6 mice transgenic for human SerpinB3 andin hepatoma cells transfected with plasmid carrying the SerpinB3gene. Moreover the inhibitory effect of SerpinB3 on calpain wasassessed by means of the calpain activity assay.

Results A positive correlation between Myc and SerpinB3expression was observed at transcription and protein level in HCCspecimens, where Myc oncogene was found predominantly in thenucleus of cancer cells overexpressing SerpinB3. Myc expressionwas significantly up-regulated by SerpinB3 through calpain andHyppo-dependent molecular mechanisms. Recombinant SerpinB3protein was indeed capable to inhibit the activity of calpain invitro, likely reducing its ability to cleave Myc in its non-oncogenicMyc-nick cytoplasmic form. Furthermore, SerpinB3 increased thetranscription of Myc through the induction of Yap pathway, asdocumented by a remarkable Yap nuclear translocation and up-regulation of Yap target genes.

Conclusions Data from the present study provide evidence thatSerpinB3 can improve the production of Myc oncogene through
direct and indirect mechanisms that include the inhibition of gen-eration of its cytoplasmic form and the activation of Yap pathway.

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-28

RANSARTERIAL CHEMOOCCLUSION (TACO)ITH DEGRADABLE-STARCH-MICROSPHERE IN

NRESECABLE HEPATOCELLULAR CARCINOMA:PROSPECTIVE PILOT-STUDY

. Francioso, A. Orlacchio, F. Chegai, F. Santopaolo,. Lenci, M. Milana, A. Brega, M. Angelico

Liver and Transplant Unit, Tor Vergata University,ome, Italy

Background: According to BCLC algorithm, Transarterialhemoembolization is recommended for unresecable hepatocellu-

ar carcinoma (HCC) in patients with preserved liver function. Thentraarterial chemotherapy with transient artery occlusion coulde a valid treatment alternative in patients with high-risk of liverailure.

Aim of the present study was to evaluate efficacy and safetyf TACO using Degradable-Starch-Microspheres (DSM) in patientsith unresecable HCC.

Methods We prospectively enrolled 24 cirrhotic patients21/3 M/F, mean age 66.3 ± 10.5 years) with unresecable HCC, toe treated with three consecutives DSM-TACO at 5-week inter-als. Chemotherapy drug was Doxorubicin Chloridrate (50 mg/m2)ixed with 275 mg DSM (Embocept®S, PharmaCept). Liver func-

ion before and after treatments was evaluated with Child Pugh andELD scores. Treatment response was assessed by CT-scan 4 weeks

fter procedures according to mRECIST criteria.Results: Overall, Complete Response (CR) was observed in 5/24

20.8%), 9/24 (37.5%) and 14/24 (58.3%) pts after the first, the secondnd the third procedure, respectively. No patient showed Stableisease or Disease Progression at the end of the study. Patientsith monolobar disease (14/24: 58.3%) showed significant CR after

he 1st procedure compared with bilobar HCC localization (5/14s 0/10; p = 0.017). At the end, CR did not differ between monor bilobar disease (9/14: 64.2% vs 5/10: 50%; p = ns). Drop-outsccurred in 8 patients (33.3%): 4 after the 1st treatment (3 wors-ned liver function, 1 liver transplanted), 4 after the 2nd treatment2 worsened liver function, 1 liver transplanted, 1 non compliance).ostembolization syndrome was observed in 9 (37.5%), in 4 (16.6%)equiring extended hospitalization. One patient had cholecystitisesolved with medical care.

Conclusions: DSM-TACO offers a valid TACE alternative inatients with unresecable HCC. Careful patients selection isequired to avoid liver failure in patients with border-line liver
ompensation. Further investigation to define DSM-TACO effectsn survival is warranted.

sease 47S (2015) e19–e42

T-29

HIF2� NEDDYLATION AS A SELECTIVESERPINB3-DEPENDENT MECHANISM LEADINGTO ITS INCREASED STABILIZATION ANDNUCLEAR TRANSLOCATION IN LIVER CANCERCELLS

S. Cannito 1, G. Villano 2, C. Turato 2, E. Morello 1,C. Paternostro 1, E. Novo 1, S. Quarta 2,S. Colombatto 3, F. Lopitz-Otsoa 4,M.L. Martínez-Chantar 4, P. Pontisso 2, M. Parola 1

1 Department of Clinical and Biological Sciences, Unitof Experimental Medicine, University of Turin, Turin,Italy2 Department of Medicine, University of Padua,Padua, Italy3 Department of Oncology, University of Turin, Turin,Italy4 Department of Metabolomics, CIC bioGUNE, Centrode Investigacion Biomedica en Red de EnfermedadesHepaticas y Digestivas (Ciberehd), Technology Parkof Bizkaia, Bizkaia, Spain

Background SerpinB3, a cysteine-proteases inhibitor overex-pressed in hepatocellular carcinoma (HCC), has been reported tobe up-regulated by hypoxia through a HIF2�-dependent mecha-nism and to act as a paracrine mediator able to induce stabilizationand increased nuclear translocation of HIF2� and up-regulationof HIF2�-related genes in liver cancer cells. The ubiquitin-likemolecule NEDD8 is a key regulator of cell growth, viability andmalignant transformation and neddylation promotes stabilizationof proteins with essential regulatory roles. Overexpression of globalneddylation has been detected in HCC and suggested to be associ-ated with its poorest prognosis.

Aims To investigate whether SerpinB3 may induce selectiveHiF2� neddylation/stabilization in liver cancer cells.

Methods The cross-talk between SerpinB3 and HIF2� has beeninvestigated by taking advantage of morphological, molecular andcell biology techniques in the following experimental models: i)control HepG2 exposed to recombinant SerpinB3 (rSerpinB3); ii)HepG2 stably transfected to overexpress SerpinB3 (HepG2/SB3);iii) transgenic mice overexpressing SerpinB3 in the liver.

Results Immunohistochemistry performed onliver sectionsfrom SerpinB3 transgenic mice shows an impressive level of nuclearstaining for HIF2� and exposure of HepG2 to rSerpinB3 results inincreased HIF2� nuclear levels. Different experimental approachesrevealed that in relation to the action of SerpinB3: increased HIF2�protein levels are i) unrelated to its increased transcription as wellas ii) to an inhibition of overall proteasome activity by SerpinB3;iii) overexpression of SerpinB3 leads to an increase in the levelof neddylated proteins and of levels of NEDD8 activating enzyme1 (NAE-1); iv) the use of a specific inhibitor of NAE-1 selectivelyreduces HIF2� levels.

Conclusions SerpinB3 can affect the behaviour of target cells byincreasing protein levels of HIF2�, followed by its nuclear translo-cation and induced transcription of target genes, by inducing HIF2�
selective neddylation and consequent stabilization which is inde-pendent on hypoxia.

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-30

-DIMER AND FIBRINOLYTIC ACTIVITY INATIENTS WITH DECOMPENSATED LIVERIRRHOSIS

.G. Romanelli a, A.P. Cellai b, D. Lami b,. Natucci a, C. Tosti-Guerra a, R. Abbate b,. Prisco b, G. Laffi a

Dipartimento Medicina Clinica e SperimentaleDMSC) Liver Unit, ItalySezione delle Malattie Aterotrombotiche delipartimento dell’Area Critica Medico Chirurgicaniversità di Firenze - Azienda Ospedalieroniversitaria Careggi (AOUC), Firenze, Italy

Background and Aims: Cirrhotic plasma could generate simi-ar or even greater amount of thrombin; a procoagulant imbalanceas been introduced. Ascitic fluid has been hypothesized to be therigin of hyperfibrinolysis. Standard tests (INR) fail to really reflectleeding tendency. Aim of this study was to determine whether abrinolytic activity is detectable in ascites.

Material andMethods: We evaluated 33 patients with liver cir-hosis (11 in Child-Pugh class A, score 5.6, mean age 65 ± 11 (yrs);

in Child-Pugh class B, score 7.9, mean age 69 ± 17 (yrs); 13 inhild-Pugh class C, score 11.4, mean age 70 ± 12 (yrs) and 21 controlealthy subjects, mean age 68 ± 14 (yrs). We studied Clot Lysis TimeCLT), D-dimer, tissue plasminogen activator (t-PA), plasminogenctivator inhibitor (PAI-1), alpha2-antiplasmin (a2AP), plasmino-en (PLG), thrombin activatable fibrinolysis inhibitor (TAFI), andthrough EuroCLOT) clot formation, structure, and lysis.

Results: We found that a2AP (%), PLG (%), TAFI (mg/mL) andbrinogen (mg/dl) levels were significantly higher in plasma thanscites (p < 0.001) and lower in plasma from patients than con-rols (p < 0.05). Instead, D-dimer levels (ng/ml) were significantlyower in plasma than in ascitic fluid (p < 0.001), whereas similaroncentrations were found for t-PA and PAI-1. D-dimer, PAI-1 and-PA levels were significantly higher (p < 0.05) and CLT shorter inlasma from patients than from healthy subjects (p < 0.05). By Euro-LOT, statistically significant differences were observed betweenirrhotic patients and healthy controls (AUC control subjects: 585;hild-Pugh A 316*; Child-Pugh B 257*; Child-Pugh C 129*** (from< 0.05* to p < 0.001***).

Conclusion: Cirrhotic patients show hyperfibrinolytic activityersus healthy subjects. In ascitic fluid from cirrhotic patients highevels of D-dimer were found. These data further suggest that asciticuid compartment is in continuous exchange with plasmatic com-
artment, possibly through lymphatic flux, and contributes to theoagulopathy of liver cirrhosis.

sease 47S (2015) e19–e42 e33

T-31

SURVIVAL OF PATIENTS WITH HEPATOCELLULARCARCINOMA (HCC) WITHIN THE BOLOGNA LIVERONCOLOGY GROUP: COMPARISON WITHINTERNATIONAL GUIDELINES

E. Terzi, M. Piccinnu, F. Piscaglia, S. Leoni,A. Granito, L. Bolondi, on the behalf of theBLOG-Bologna Liver Oncology Group

Division of Internal Medicine, Department ofMedical and Surgical Sciences, University of Bologna,Bologna, Italy

Introduction: The Barcelona Clinic Liver Cance system (BCLC),endorsed by the latest international EASL-EORTC and AASLD guide-lines, represents the reference therapeutic and prognostic stagingsystem.

Aim: was to evaluate the overall survival and clinical deter-minants of survival in our series of HCC patients and to comparethe results to those of guidelines eventually attempting to identifyreasons for discrepancy.

Materials and Methods: Among 1028 HCC patients referred toour center between January 2000 and August 2013, we retrospec-tively identified the outcome of the 595 consecutive patients seenin our center on occasion of the first diagnosis of HCC.

Results: Stage was BCLC-0 in 23 patients (4%), BCLC-A in 273(46%,), BCLC-B in 155 (26%), BCLC-C in 114 (19%) and BCLC-D in 30patients (4%). Median survival in BCLC-0 was 88 months (95% C.I.41.3-134.7) with 1-, 3- and 5-year survival rates of 97%, 86% and61%; in BCLC-A was 44 months (95% C.I. 37.4-50.4) with 1-, 3- and5-year survival rates of 92%, 60% and 36%; in BCLC-B was 20 months(95% C.I. 14.2-25.8) with 1-, 3- and 5-year survival rates of 65%, 34%and 21%; in BCLC-C was 8 months (95% C.I. 5.2-10.8) with 1-, 3- and5-year survival rates of 39%, 2% and 0%; in BCLC-D was 7 months(95% C.I. 4.8-12.2). At multivariate survival analysis, age > 67 years,neoplastic portal vein thrombosis, AFP > 19 ng/mL and BCLC C-Dwere statistically associated with shorter survival.

Conclusion: The present study validates survival data reportedby the guidelines in a large unselected series of consecutive patientswith HCC managed under real life conditions. Such results wereachieved through a multidisciplinary and individually tailoredtreatment strategy of HCC patients within the Bologna Liver Oncol-ogy Group (BLOG).


T-32

TACE WITH CONE BEAM COMPUTEDTOMOGRAPHY IS MORE EFFECTIVE THANTRADITIONAL TECHNIQUE IN BCLC A HCCPATIENTS INELIGIBLE TO SURGERY

R. Patti 1,2, N. Mezzina 1,2, F. Masutti 1,2,C. Abazia 1,2, V. Lanzillotti 1,2, D. Pascut 2,C. Sukowati 2, F. Pozzi Muceli 3, C. Tiribelli 1,2,S.L. Crocè 1,2

1 Dipartimento di Scienze Mediche, Clinica Patologiedel Fegato, Università di Trieste, Trieste2 Fondazione Italiana Fegato, Italy3 Dipartimento di Scienze Mediche, UCO diRadiologia, Università di Trieste, Trieste

Introduction: In the last years despite the improvement in can-cer diagnostic and therapeutic tools HCC mortality is still high. Thedecision-making process in HCC treatment is mainly based on the

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arcelona Clinic Liver Cancer (BCLC) staging system that consideroth clinical and tumor characteristics. BCLC A patients shouldndergo surgical ablation but they are often not eligible due toomorbidities or tumor position. So the only treatment availables the trans-arterial-chemoembolization (TACE) performed either

ith traditional radiologic procedures or with Cone Beam Com-uted Tomography (CBCT) that allow a better 3D reconstruction ofhe HCC nodule and its supplying vessels.

Aim: Our study aims to evaluate the efficacy of TACE performedith CBCT or with traditional tecniques in the subpopulation ofatient in BCLC A ineligible to surgery.

Methods: Between January 2012 and June 2014 32 patientsere included in the study and 54 procedures were performed.

atients in BCLC A treated with CBCT were 57% (12/21) and patientreated with traditional technique were 27% (9/33).

Diagnosis of HCC is done considering EASL criteria and the radio-ogic response was evaluated at one month after treatment with theECIST score.

Results: The two groups were homogenous with no statisticallyifferences in their characteristics.

Patients in the CBCT group show a better response than patientsn traditional group (a 83% vs. 36%, p = 0.029). Patients in BCLC, instead, have similar response irrespective to the treatmenteceived (44% vs 54, CBCT and traditional group respectively, NA).

Conclusion: In BCLC A patients TACE performed with CBCT isignificantly more effective that the conventional procedure. This isrobably due to a more precise identification of the small nodule(s)nd blood supply. When the lesion is lager (group B) the advantagef CBCT disappears.


-33

LINICALLY GUIDED ADJUSTMENT OFORAFENIB DAILY DOSE IS ADVISABLE FORIRRHOTICS PATIENTS WITH HEPATOCELLULARARCINOMA (HCC)

. Gallusi, A. De Santis, C. Iegri, M. Lupo,

. Mascolo, A.F. Attili

Department of Gastroenterology, La Sapienzaniversity of Rome, Rome, Italy

Introduction: Sorafenib is approved for treatment of advancedCC. Its efficacy/tolerability is often unpredictable. We looked forrognostic factors of survival in a population of patients assumingorafenib for HCC.

Patients and methods: Ninety-three cirrhotic patients with HCCere prospectively followed-up until last clinical control/death.ommonest patients’ characteristics were: male sex (79,6%); viraltiology (HCV/HBV) of cirrhosis (51,1%); ECOG PS 0 (68,9%) vs 126,7%) and 2 (4,4%); BCLC C (60,4%) vs B (39,6%); presence of: por-al vein thrombosis (57,8%), esophageal varices (57,1%), metastasis76,7%). Median age was 70 (45-88; 95% CI 6472); median Child-ugh score was 6 (range 5-9; 95% CI 5,79-6,20). Everybody startedherapy at 800 mg/day. If needed, posology was then reducedccording to patient’s tolerability and/or adverse events. Modifica-ions of the posology/interruptions of treatment were registered. Athe end of observation, the dose assumed for ≥60% of the treatmenteriod was considered representative for each patient.

Results: In the entire population, median overall survival was85 days (95% CI 161-256); median treatment duration 133 days
95% CI 103-182). Skin toxicity (hand-foot skin reaction/rash) washe most common adverse event (41,9%) and was associated to aignificantly higher median survival (396 vs 206 days, p = 0,02).
sease 47S (2015) e19–e42

Fifty-four patients (58.1%) assumed sorafenib 800 mg/day and 39patients (41.9%) 400 mg/day. The two groups did not differ forthe variables listed above. Probability of survival was significantlyhigher with daily dose of 400 mg than 800 mg (p = 0,000488) (Fig-ure 1). Accordingly, prevalence of skin toxicity was significantlyhigher in patients treated with 400 mg/day than 800 mg/day. Atmultivariate analysis, sorafenib dose and skin toxicity resultedthe only variables significantly associated to survival (respectivelyp = 0,0009 and p = 0,0245).

Conclusions: When guided by clinical judgment of opportunity,dose reduction should be considered feasible and maybe advisable,especially in case of skin toxicity.


T-34

ANALYSIS OF THE ASSOCIATION OF PEDIATRICNAFLD WITH TWO SNPS OF GENES ENCODINGFOR FIBROBLAST GROWTH FACTORS 19 AND 21RECEPTOR SYSTEM

Crudele, S. Ceccarelli, N. Panera, D. Gnani,C. De Stefanis, A. Alisi, V. Nobili

Liver Research Unit, Bambino Gesù Children Hospitaland IRCCS, Rome, Italy

Introduction: Non-alcoholic fatty liver disease (NAFLD) is oneof most common liver diseases worldwide characterized by alarge spectrum of histological damages, including simple steato-sis (NAFL), necro-inflammation and non-alcoholic steatohepatitis(NASH), eventually associated with liver fibrosis. It is known thatNAFLD is a multifactorial disease caused by the interaction betweenenvironmental factors and genetic background that trigger themultiple mechanisms involved in disease pathogenesis. Fibroblastgrowth factor(FGF)19 and FGF21 have been recently associated topediatric NAFLD suggesting a crucial role of their receptor system.

Aim: The aim of the present study was to investigate thepotential association of NAFLD with two single nucleotide polymor-phisms (SNPs) of two genes encoding for proteins that compose theFGF19/21 receptor system: the variant rs17618244 (Arg728Gln) ofKlotho-beta co-receptor (KLB) and the variant rs1966265 (Val10Ile)of fibroblast growth factor receptor 4 (FGFR4).

Materials and Methods: To this purpose, we performed a case-control study including 100 Italian children without NAFLD and150 with biopsy-proven NAFLD (74 with NAFL and 76 with NASH).The SNPs were evaluated by a 5′-nuclease TaqMan assay.

Results: In the case-control study, the presence of NAFLDincreased with the number of G alleles for rs17618244 variant inKLB (p < 0.01) and for rs1966265 variant in FGFR4 (p < 0.001). InNAFLD children the severity of the disease (measured in termsof the presence of NASH) was associated with several anthropo-metric and metabolic parameters of patients (body mass index,cholesterol, transaminases and triglycerides), but not with a higherpresence of rs17618244 G allele. Interestingly, NAFLD children car-rying the rs1966265 A allele had a trend for a higher prevalence ofNASH.

Conclusions: Our results suggest that the variant rs17618244 inKLB and the variant rs1966265 in FGFR4 could be negatively asso-

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-35

ELATIONSHIP BETWEEN INFLAMMATORYEDIATORS AND FREQUENCY OF THE INNATE

MMUNE CELLS IN PATIENTS WITHON-ALCOHOLIC FATTY LIVER DISEASE

. Maggio 1,3, L. Antonucci 1, C. Viscomi 1,. Costantini 2, G. Castello 2, C. Balsano 1,3

Francesco Balsano Foundation, Rome, ItalyCentro Ricerche Oncologiche di Mercogliano,

stituto Nazionale Per Lo Studio E La Cura Dei TumoriFondazione Giovanni Pascale”, IRCCS, MercoglianoAV), ItalyInstitute of Institute of molecular biology andathology (IBPM), National Research Council (CNR),ome, Italy

Nonalcoholic fatty liver disease (NAFLD) is the main liver dis-aseworldwide. Simple steatosis remains a benign process, even ift may progress to non alcoholicsteatohepatitis with necroinflam-

atory activity and inflammatory cell infiltration. Several studiesave demonstrated the involvement of proinflammatorymedia-ors and innate immune cells in the NAFLD pathogenesis. Wenvestigated a possible relationship between circulating levels ofro-inflammatory cytokines and percentage of immune cellular

nnate components in NAFLD patients.We enrolled 30 adult patients with a clinical diagnosis of

AFLD and 10 healthy donors. The analysis ofimmune mediatorsas performed on serum samples using multiplex immunoas-

ays.Peripheral blood mononuclear cells were isolated fromatients and circulating �� T, NK, and NKT cells weremonitored byow cytometry. Correlation analysis was performed using Pearsonorrelation test.

NAFLD patients showed elevated serum levels of chemoattrac-ant proteins IL-8/CXCL8 and MIP1-�/CCL4,compared to normalubjects (p < 0.05). Levels of IL-8/CXCL8 correlated significantlyith MIP1-�/CCL4levels, indicating a coordinated regulation of

heir secretion (p = 0.001, r = 0.723). We observed a strong posi-ivelinear correlation between IL-8/CXCL8 serum levels and �� Tell-frequency (p = 0.001, r = 0.716). Moreover, IL-8levels were sig-ificantly associated with the percentage of NKT cells (p = 0.003,= 0.773), while nocorrelation was observed with the percentage ofK cells. We revealed a markedly positive relationship between theIP1-�/CCL4 serum levels and the frequency of �� T cells (p = 0.002,

= 0.720). Thesame trend was obtained between MIP1-�/CCL4evels and the percentage of NKT cells (p = 0.005,r = 0.627).In con-lusion, we demonstrated a direct correlation between the levelsf the pro-inflammatory cytokines IL-8/CXCL8 and MIP1-�/CCL4,nd the cellular frequency of the innate immune system in NAFLDatients. Thismight suggest that the levelsof the pro-inflammatory
ytokines investigated could represent a “substratum” for themmunological events occurring during the NAFLD progression.

sease 47S (2015) e19–e42 e35

T-36

THE USE OF A POCKET-SIZED ULTRASOUNDDEVICE (PUD) IMPROVES PHYSICALEXAMINATION: RESULTS OF AN IN- ANDOUTPATIENT STUDY

A. Colli 1, D. Prati 2, M. Fraquelli 3, S. Segato 4,P.P. Vescovi 5, F. Colombo 6, C. Balduini 7,A. Baccarin 3, S. Della Valle 8, D. Conte 3,G. Casazza 9, for the Lombardy Ecoscopy Project

1 Dipartimento Medicina Interna, Ospedale AManzoni, Lecco, Italy2 Dipartimento Medicina Trasfusionale e Ematologia,Ospedale A Manzoni, Lecco, Italy3 Unità di Gastroenterologia ed Endoscopia, IRCCSFondazione Ca Granda Ospedale MaggiorePoliclinico, Università degli Studi di Milano, Italy4 Unità di Gastroenterologia ed Endoscopia, AziendaOspedaliero Universitaria Macchi, Varese, Italy5 Divisione di Medicina Interna, Azienda Ospedaliera“Carlo Poma” Mantova Italy6 Prima Divisione di Medicina Interna, A.O.NiguardaMilan, Italy7 Terza Divisione di Medicina Interna, FondazioneIRCCS Policlinico San Matteo-Università degli Studi,Pavia, Italy8 Dipartimento di Medicina Interna, Ospedale AManzoni Lecco, Italy9 Dipartimento di Scienze Biomediche e Cliniche,Ospedale L. Sacco, Universita degli Studi di Milano,Milano, Italy

Background/aim The performance of PUD is comparable withthat of standard ultrasonography, whereas the accuracy of physicalexamination (PE) is often poor requiring further tests to assess diag-nostic hypotheses. Adding PUD to PE could lead to an incrementalbenefit. Present cohort-impact study aimedat assessing whetherthe use of PUD in the context of PE could reduce the request rateof additional tests when used by physicians in different clinicalsettings

Methods The study involved medical wards(#4), hepatologicaloutpatient clinic(#1) and 90 GPs operating in the same geograph-ical area. After a short training course 135 physicians used PUDin addition to PE in 1962 consecutive patients to investigateten well-defined clinical hypotheses (ascites, biliary-duct dila-tion/gallstones, splenomegaly, pleural/pericardial effusion, urinaryretention/stones, abdominal mass/aortic aneurysm. PUD-relatedfindings were recorded. and the decision as to whether to requestfurther tests was left to the physician clinical judgement. The mainoutcome was to define the proportion of cases undergoing addi-tional tests after PUD. An accurate report of the frequency of theclinical questions in the different settings was also planning, usinglogistic regression analysis to assess the determinant(s) of the pri-mary outcome.

Results Of the 1962 included patients 726(37%) were inpatients,510(26%) hepatology outpatients and 726(37%) recruited from GPs.Gallstone (37%), ascites (17%), pleural effusion (13%), urinary stones(13%) and urinary retention(12%) accounted for > 90% of the clinicalquestions, confirmed by PUD in 66% of cases. The overall frequencyof further tests after PUD was 37%; and logistic was associated withboth the clinical questions and settings(p < 0.01). The rate of agree-
ment between PUD findings and additional tests was 89% (Sens91%,Spec 83%,LR + 5.4;LR-0.11).
Conclusions After a short training, PUD examination can beused in addition to PE in in- and outpatients to improve the answer

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-37

DHERENCE TO EASL-EORTC CLINICALUIDELINES FOR THE MANAGEMENT OFEPATOCELLULAR CARCINOMA IN FIELDRACTICE: RESULTS FROM THE ITALICAATABASE

. Sacco, V. Mismas, L. Giacomelli, S. Marceglia,

. Romano, M. Bertini, M. Bertoni, G. Federici,

. Parisi, S. Metrangolo, E. Tumino, G. Bresci

Department of Gastroenterology, Pisa Universityospital, Pisa, Italy

Introduction: Data on adherence to joint guidelines for theanagement of hepatocellular carcinoma (HCC) published in 2012

y the European Association for the Study of the Liver (EASL) andhe European Organization for Research and Treatment of CancerEORTC) are lacking.

Aim: We retrospectively evaluated the adherence to EASL-ORTC guidelines in field-practice, using data from HCC patientsegistered in the Nation-wide Italian database ITA.LI.CA. and diag-osed from 2012.

Methods: The ITA.LI.CA. database contains data of 5428 HCCatients treated at 18 Italian Centers. Patients were stratifiedccording to Child-Pugh (CP) and and the Barcelona Clinic Liverancer (BCLC) classifications. We investigated the adherence tourveillance, diagnosis, and first-line treatment recommendations.

Results: In ITALICA, 600 patients were diagnosed of HCC since012 (466 males; mean ± SD age 67.4 ± 10.9 years; 277(46.2%)P-A and 163(27.2%) CP-B; 44(8%) BCLC-0, 193(35.1%) BCLC-A,3(16.9%) BCLC-B, 172(31.3%) BCLC-C, 48(8.7%) BCLC-D). Over-ll, 317(55.2%) were diagnosed during a surveillance program. Ofhem, 231(57.9%) were cirrhotic (median surveillance duration: 6

onths). Four-hundred-ninety-six (85.3%, 449 cirrhotic) patientsere diagnosed applying a radiological, 80(13.7%) a histological,

nd 6(1%) a cytological criterion. Five (9.7%) patients in BCLC stage 0ith CP A, and single nodules underwent tumour resection; 3(1.4%)atients in BCLC-A received liver transplantation, and 83(43.1%)eceived radiofrequency ablation or Percutaneous Ethanol Injec-ion. Intermediate HCC-stage patients (BCLC-B) receiving TACEere 45(47.9%), and advanced-stage patients (BCLC-C) receiving

orafenib were 38(21.9%). Palliative care was provided to terminaltage patients (BCLC-D) in 31(64.3%) cases.

Conclusions: The overall adherence in a “real-world” practice toASL-EORTC guidelines was low, particularly in patients with earlytage HCC. Difficulties inpatients staging and the high prevalence oflder patients with relevant co-morbidities may partially explainhese findings. Strategies to help improve adherence to interna-
ional guidelines for HCC in field-practice and new scoring criteriare required.

sease 47S (2015) e19–e42

T-38

CLINICAL PATTERNS OF HEPATOCELLULARCARCINOMA (HCC) IN NON ALCOHOLIC FATTYLIVER DISEASE (NAFLD): A MULTICENTERCASE-CONTROL STUDY

F. Piscaglia 1, G. Svegliati Baroni 2, A. Barchetti 3,A. Pecorelli 1, S. Marinelli 1, C. Tiribelli 3,S. Bellentani 3 e gruppo di studio italiano HCCNAFLD composto da: L. Bolondi, M. Zoli,D. Malagotti, G. Brandi, E. Bugianesi, E. Vanni,L. Mezzabotta, G. Cabibbo, S. Petta, A. Fracanzani,S. Fargion, F. Marra, B. Fani, R. Sacco, F. Morisco,N. Caporaso, M. Guarino, M. Colombo,R. D’Ambrosio, L.S. Crocè, R. Patti, E. Giannini,A. Lonardo, E. Baldelli, L. Miele, A. Grieco,F. Farinati, C. Pozzan, M. Borzio, E. Dionigi,G. Soardo, P. Roselli, F. Ciccarese, F. Virdone,A. Affronti, F.G. Foschi, F. Borzio, F. Trevisani

1 Unità di Medicina Interna, Dipartimento di ScienzeMediche e Chirurgiche, Policlinico S.Orsola-Malpighi, Università of Bologna2 Unità di Gastroenterologia, Università Politecnicadelle Marche, Ancona3 Centro Studi Fegato (CSF), Liver ResearchCenter–Parco della Scienza - Basovizza Campus,Trieste

Background The prevalence of non-alcoholic fatty liver disease(NAFLD) is growing and the disease can progress to hepatocellularcarcinoma (HCC). Only scant clinical information on HCC in NAFLDis available.

The aim of the study was to assess the clinical features of patientswith NAFLD-related HCC (NAFLD-HCC), and to compare them tothose having HCV-related HCC.

Methods: The study is a multicenter observational case-controlstudy. 756 patients with either NAFLD (145) or HCV-related chronicliver disease (611) have been enrolled in Secondary Care ItalianCenters. Survival was modeled according to clinical parameters,lead time bias and propensity analysis.

Results: Patients with NAFLD- HCC were: less often male thanHCV-HCC (61% vs 79%), had a higher BMI (29.1 vs 27.6), similarChild-Pugh score (5.6 vs 5.8), larger tumor volume (largest nodule4.1 vs 3.3 cm), more often infiltrative pattern (15.4% vs 4.0%) anddetection outside specific surveillance (52.4% vs 36.7%). Survivalwas significantly shorter (p = 0.017) in NAFLD-HCC: 25.5 months(95% CI 21.9-29.1) than in HCV-HCC patients 33.7 months (95% CI31.9-35.4). This difference remained significant even after adjust-ment for lead time bias in surveilled patients. Cirrhosis was presentin only about 50% of NAFLD-HCC patients. To understand the intrin-sic aggressiveness of HCC and to eliminate possible confounders, apropensity score analysis was carried out considering age, gender,

group. The difference in mean surviva between the two groups wasno longer statistically significant (30.2 months in NAFLD-HCC and36.9 HCV- HCC, p = 0.330).

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Conclusions NAFLD-HCC rises in the absence of cirrhosis inbout 50% of the cases, and it is detected at a later tumor stage thann HCV-infected patients. Future research should identify patients

ith NAFLD who require surveillance in order to offer them theost timely and effective treatment.


-39

AS-LIGAND INVOLVEMENT IN END STAGE LIVERISEASE

. Trombetta 1, P. Cetrangolo 1, A. Cattaneo 1,. Rossi 2, D. Prati 3, L. Porretti 1, F. Colombo 1,∗

Flow Cytometry Service, Fondazione IRCCS Ca’randa Ospedale Maggiore Policlinico di Milano,ilan, ItalyLiver Transplant Unit, Fondazione IRCCS Ca’ Grandaspedale Maggiore Policlinico di Milano, Milan, ItalyDepartment of Transfusion Medicine andaematology, Ospedale “A. Manzoni”, Lecco, Italy

Introduction: FasL has been involved in lymphocyte functionnd disease development. Recently, it has been demonstrated thatD8 + /FasL + cells can be eliminated by tumor cells as escape mech-nism.

Aim: To evaluate if CD8 + /FasL + lymphocytes are also involvedn end stage liver disease.

Materials and Methods: 6 blood donors and 4 cirrhotic patientsere enrolled. FasL expression on CD8 + cells of donors and patientseripheral blood (PB) was evaluated by flow cytometry beforend after 24 h of activation with PMA and ionomycin. Moreover,e evaluated FasL expression on CD8 + cells infiltrating patients’

iver parenchyma in bioptic specimens. To mimic lymphocyte-liverarenchyma interaction, we implemented co-cultures with lym-hocyte and HCC cell lines. Donor PBMC, either before or after 4 h ofctivation, were co-cultured with 2 FasL + HCC cell lines (ratio 1:5,8 h) or alone in the same conditions. Cell mortality was evaluatedy means of 7AAD and Zombie Aqua staining.

Results: The percentage of PB CD8 + cells was equal betweenatients and controls, while FasL expression on CD8 + cells was

ower in patients (34.2 ± 4.5 vs. 51.4 ± 5.9, p < 0.01). Similar dataere obtained after 24 h of activation, although this process did not

ncrease FasL expression on CD8 + cells. Interestingly, liver infiltrat-ng CD8 + cells expressed less FasL in comparison to the circulatingounterpart (17.5 ± 9.5 vs. 34.2 ± 4.5, p = 0.032), although no dif-erences in CD8 + percentage were observed between PB and liverarenchyma (18.4 ± 13.2 vs. 33.4 ± 4.8, p > 0.05). Finally, an equalD8 + /FASL + cell mortality was observed when resting CD8 + cellsere co-cultured with HCC cell lines (13.3 ± 3.0, 12.7 ± 4.1 vs.

0.3 ± 7.9), while co-cultures with activated CD8 + cells produced aigher CD8 + /FASL + cell death (47.1 ± 12.5, 43.9 ± 5.2 vs. 25.1 ± 8.3,< 0.05).

Conclusions: Our data suggest that diseased liver cells coulde responsible for CD8 + /FasL + depletion by an escape mechanism,
oth within the liver parenchyma and in PB, favouring the progres-ion of the disease.

sease 47S (2015) e19–e42 e37

T-40

NEUTROPHIL FUNCTION IN PATIENTS WITHCHRONIC HEPATITIS C (CHC) UNDERGOINGTRIPLE ANTIVIRAL THERAPY (TT) WITH FIRSTAND SECOND GENERATION PROTEASEINHIBITORS (PI)

M. Gambato 1,2, N. Caro-Pérez 1, N. Canete 3,Z. Marino 1, S. Lens 1, JM. Sánchez-Tapias 1,J. Carrion 1, S. Pérez-del-Pulgar 1, M. Juan 4,M. Londono 1, X. Forns 1

1 Liver Unit, Hospital Clínic, IDIBAPS, Ciberehd,Barcelona, Spain2 Multivisceral Transplant Unit, Department ofSurgery, Oncology and Gastroenterology,PaduaUniversityHospital, Padua, Italy3 Digestive disease Unit, Hospital del Mar, Barcelona,Spain4 Immunology Unit, Hospital Clinic Barcelona, Spain

Introduction Cirrhotic patients undergoing TT with peg-IFN/RBV and a first generation PI have higher risk of bacterialinfections as compared to those receiving IFN/RBV (Londono et al. JHepatol 2014). PIs might inhibit neutrophil proteases and Toll-likereceptor (TLR) pathways.

Aim We aimed at evaluating innate immune recognition andneutrophil activity in CHC patients undergoing TT.

Materials and Methods: CHC patients undergoing TT after alead-in phase were prospectively studied. Blood samples wereobtained at baseline, week 4 and week 8. Phagocytic and oxida-tive burst activities were quantified by flow cytometry. Neutrophilswere isolated from peripheral blood and stimulated with flagellin(FLA) to determine cytokine secretion (IL-1b, IL-6, IL-10, IL-12).Quantification was performed by Luminex®.

Results: 18 patients have been enrolled so far (4 F2, 3 F3 and 11F4). PIs used were telaprevir (n = 9), boceprevir (n = 1) or simeprevir(n = 8). The number of phagocytized bacteria per cell, determinedby the mean fluorescence intensity (MFI), significantly decreasedat weeks 4 and 8 as compared to baseline (p = 0.02 and p = 0.035,respectively). However, there were no significant differences in MFIbetween weeks 4 and 8. Oxidative burst capacity did not changethroughout the study period. Cytokine levels measured after neu-trophil stimulation with FLA were increased at week 4 and highlydecreased at week 8 of therapy, but the difference was not statisti-cally significant (p = 0.3 for all cytokines).

Conclusions: Antiviral therapy with peg-IFN/RBV is associatedwith a significant decrease in neutrophil phagocytic capacity, butthe addition of a PI does not seem to further reduce this function.Interestingly, after 4 weeks of PI therapy, there was a decrease incytokine secretion which might suggest an inhibition of proteasesinvolved in TLR signaling pathways. The combination of both effects

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-41

MALL FIBERS PERIPHERAL NEUROPATHY INILSON’S DISEASE: AN IN VIVO

OCUMENTATION BY CORNEAL CONFOCALICROSCOPY

.C. Sturniolo 1, O. Bartolo 1, D. Lazzarini 2,. Berton 2, A. Leonardi 2, I.A. Fregona 2,

. Midena 2

Department of Gastroenterology, University ofadua, Padua, ItalyDepartment of Ophthalmology, University ofadua, Padua, Italy

Wilson disease (WD) is an inherited autosomal recessive disor-er of copper metabolism. Corneal confocal microscopy (CCM) isfast technique to analyze the human cornea in vivo. We aimed

o investigate central corneal changes and to assess the parame-ers of corneal subbasal nerve plexus (CSNP) in patients affectedy WD, using CCM. 24 patients affected by WD and 24 healthyontrol subjects were enrolled in this cross-sectional comparativetudy. One eye of each subject was examined to quantify differentorneal parameters, by CCM. WD induced significant alterationsn both corneal subbasal nerve plexus, and corneal epithelium. Allhe parameters of the subbasal nerve plexus were altered in WD,ocumenting, for the first time, a (corneal) peripheral nerve dam-ge in WD. The decrease of major CSNP parameters confirms theamage (and death) of a significant number of small nerve fibers,hereas the increase of fiber tortuosity is a sign of tentative nerve

egeneration. Mean epithelial cell diameter (P < 0.0001) and meanpithelial cell density (P < 0.0001) resulted significantly higher andower compared to controls, respectively. No significant differencen corneal stroma and endothelium were observed; no significantifferences were documented among different treatment groups.he comparison between predominantly neurological and predom-nantly hepatic presentation didn’t show any statistical differences.he only difference in subgroup was found between patients withrevious of KF ring e without KF ring. CCM showed significantorneal changes in subbasal nerve plexus, and secondary cornealpithelium changes in WD. CCM clearly documented for the firstime, a (corneal) peripheral nerve damage in WD. Corneal damageas more severe in pts with previous KF ring. CCM may contribute

o the diagnosis and monitoring of the peripheral nervous system
nvolvement in WD. Further larger studies needed to confirm thesendings.

sease 47S (2015) e19–e42

T-42

HEPATIC VENOUS PRESSURE GRADIENT IN THEPREOPERATIVE ASSESSMENT OF PATIENTS WITHRESECTABLE HEPATOCELLULAR CARCINOMA

A. Cucchetti 1, M. Cescon 1, F. Piscaglia 1,R. Golfieri 2, M. Renzulli 2, C. Mosconi 2,A. Cappelli 2, F. Mazzotti 1, F. Neri 1, G. Ercolani 1,A.D. Pinna 1

1 Department of Medical and Surgical Sciences,S.Orsola-Malpighi Hospital, Alma Mater Studiorum -University of Bologna, Bologna, Italy2 Department of Diagnostic Medicine andPrevention, S.Orsola-Malpighi Hospital, Alma MaterStudiorum - University of Bologna, Bologna, Italy

Background: According to guidelines, hepatic resection (HR) forhepatocellular carcinoma (HCC) should be offered to patients with-out significant portal hypertension (PH), namely, with a hepaticvenous pressure gradient (HVPG) < 10 mmHg. In real life, HCCpatients with clinical signs of PH are often referred for surgicalevaluation and eventual hepatectomy.

Methods: Data from 70 consecutive, prospectively enrolled,HCC patients submitted to HR were collected and analyzed to definethe role of HVPG in the prediction of post-operative liver failure(PHLF) grade B/C, as defined by the International Study Group ofLiver Surgery.

Results: Post-operative and 90-days mortalities were null. Themedian HVPG value was 9 mmHg (range: 4-18), median MELDwas 8 (range 6-14); 34 patients had HVPG≥10 mmHg (48.6%).Forty-nine patients experienced an uneventful (Grade A) post-operative course; of them, 17 had HVPG≥10 mmHg (24.2% of70 patients). Grade B complications occurred in 20 patients (3with HVPG < 10 mmHg and 17 with HVPG≥10 mmHg; P < 0.001);only one grade C complication occurred in a patient havingHVPG < 10 mmHg, subsequently successfully submitted to livertransplantation. Receiver Operating Characteristics curve for PHLFB/C showed for Model for End-stage Liver Disease (MELD) an areaunder the curve (AUC) of 0.727 (95%C.I.: 0.598–0.857) and for HVPGof 0.792 (95%C.I.: 0.677–0.907; P = 0.475).

Conclusions: Hepatic venous pressure gradient can be usedbefore surgery to stratify the risk of PHLF but does not add sig-nificant further prognostic ability with respect to MELD score. Theproposed cut-off of 10 mmHg is indeed associated with worse prog-
nosis but excludes about one quarter of patients who will benefitfrom surgery without short-mid term post-operative sequelae.

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EPATITIS DELTA PREVALENCE AND CLINICALXPRESSION IN AN IMMIGRANT POPULATION INTALY

. Colucci 1, M. Colombo 1, G. Lunghi 2, P. Bono 2,. Fadelli 3, M. Adamoli 3, D. Aurelio 3,. Abbruzzese M 3, R. Romeo 1

Division of Gastroenterology and Hepatology,ondazione Ca’ Granda, Policlinico, Milan, ItalyLaboratory medicine Fondazione Ca’ Granda,oliclinico, Milan, ItalyPoliambulatorio Opera San Francesco per i Poveri

OSF), Milan, Italy

Background and aims: Hepatitis delta virus (HDV),a defectivegent coinfecting 5% of chronic hepatitis B virus (HBV) carriers, hasecently regained the interest of the scientific community becausef its increasing prevalence and morbidity rates, particularly ineveloping countries. In this respect, we started a screening pro-ram at one of the largest NGO clinic in Italy, Opera San FrancescoOSF) in Milan where patients are mainly illegal immigrants orefugees lacking health insurance coverage and often proper hous-ng accommodation.

Methods: Using commercially available tests (Diapro Srl, Sesto. Giovanni, Italy) HBV and HDV infections were sought in consec-tive, unselected individuals, independently on their health status.ollowing HBsAg and IgG and IgM anti-HDV detection, positiveatients underwent HBVDNA and HDV RNA testing whereas thecreening tests were offered to their close relatives.

Results: Starting September 2014, 119 migrants originatingrom North Africa (10%), Eastern Europe (48%), Central and Southmerica (33%) and Asia (9%) have been screened. 23 patientsesulted HBsAg positive of whom two patients with compensatedirrhosis tested also IgG and IgM anti-HDV positive but HDV RNAndetectable (< 30 copies/ml) and are currently treated with Ente-avir, 300 mg/die.

Conclusion: This interim analysis suggests a high HBsAg car-ier state prevalence (19.3%) and HBV/HDv co infection rate of 8.6%mong migrants slightly lower than that recently reported, partic-larly in east Europe, but probably better reflecting the situation ofhe general population in these countries as we did not apply anyatients selection. By the time this screening project is concluded itill be the largest ever carried out and the most informative on the

ctual HBsAg and HDV prevalence in a mixed immigrant populationrom high endemic countries.


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T-44

PNPLA3 VARIANT IS AN INDEPENDENTPREDICTOR OF SEVERE STEATOSIS IN PATIENTSWITH CRONIC HEPATITIS C AND HIV INFECTION

C. Sagnelli 1, M. Merli 2, C. Uberti-Foppa 2,A. Gradone 3, H. Hasson 2, G. Cirillo 3,S. Salpietro 2, C. Minichini 4,E. Miraglia Del Giudice 3, A. Lazzarin 2,E. Sagnelli 4, N. Coppola 4

1 Department Experimental Medicine and Surgery “F.Magrassi e A. Lanzara”, Second University of Naples,Naples, Italy2 Department of Infectious Diseases, Vita-SaluteUniversity,San Raffaele Scientific Institute, Milan,Italy3 Department of Pediatrics, Second University ofNaples, Naples, Italy4 Department Mental Health and Public Medicine,Section of Infectious Diseases, Second University ofNaples, Naples, Italy

Background A correlation between the p.148 polymorphismof the patatin-like phospholipase domain-containing protein 3(PNPLA3) and the degree of liver steatosis in patients with chronichepatitis C (CHC) was identified.

Aim: to analyze the impact of PNPLA3 variants on liver histologyof CHC patients with HIV infection.

Methods: 168 consecutive patients with HIV infection andbiopsy proven CHC, naïve for HCV-treatment, HBsAg-negative andwith no alcohol abuse were enrolled. Patients aged 40.7 ± 6.0, 72.6%were males, 42% with HCV-genotype 3 and 38.9% with genotype1, 81.5% in HAART; the nadir of CD4 + was 290.0 ± 197.4 cell/�L.Liver fibrosis and necroinflammation were analyzed by the Ishak’sscoring system and steatosis as follow: score 1 = fatty depositionin 1-10% of hepatocytes, score 2 in 11-30%; score 3 in 31-60% andscore 4 in > 60%.

Results: No difference was observed in baseline demographic,laboratory characteristics and degree of necroinflammation andfibrosis between patients with different PNPLA3 variants. The 79subjects with PNPLA3 I/M-M/M variants showed a severe steato-sis (degree 3-4) more frequently than the 89 with PNPLA3 I/I (43%vs 24.7%, p = 0.001). Compared with the 112 patients with steato-sis score 1-2, the 56 with score 3-4 had higher BMI (23.91 ± 2.99vs 22.88 ± 2.82, p = 0.03), ALT serum level (106.84 ± 63.11 vs.70.53 ± 60.16, p = 0.0002), higher prevalence of cases with HCV-genotype 3 (55.6% vs. 35.2%, p = 0.01) and of PNPLA3 I/M-M/M(60.7% vs 39.3%, p 0.01).

A multivariate logistic regression analysis including PNAPL3variants (I/M or M/M vs I/I), HCV genotype (3 vs. other genotypes),BMI and other potential confounding factors (age, sex) showeda high BMI (0.0165), HCV-genotype 3 (p = 0.0075) and PNPLA3p.I148 I/M-M/M polymorphism (p = 0.0088) independently associ-ated with liver steatosis score 3-4.

Conclusions: This study is the first demonstration that the
PNPLA3 p.148 I/M-M/M variants is an independent predictor ofsevere liver steatosis in CHC patients with HIV infection

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-45

ROACTIVE DOSE ADJUSTMENTS AREECESSARY IN MANY ADV-EXPERIENCEDATIENTS TREATED WITH TDF MONOTHERAPYOR 5 YEARS: A PROSPECTIVE COHORT STUDY IN20 PATIENTS

. Lampertico 1, R. Soffredini 1, M. Borghi 1,. Viganò 2, F. Facchetti 1, E. Galmozzi 1,

. Invernizzi 1, M. Colombo 1

Division of Gastroenterology and Hepatology,ondazione IRCCS Cà Granda Ospedale Maggioreoliclinico, University of Milan, Milan, ItalyDivision of Hepatology, Ospedale San Giuseppe,niversity of Milan, Milan, Italy

Introduction: Tenofovir (TDF) is a popular anti-HBV strategyor NUC-experienced patients, but its safety profile in field practiceatients previously exposed long-term to Adefovir (ADV) is underiscussion because of recent reports of Fanconi syndrome.

Materials and Methods Results: 320 NUC-experienced CHBatients received Tenofovir (TDF) for 69 months (range 1-106) asswitch from ADV + LAM (86%) or as a rescue therapy for LAM,DV or ETV (age 59, 85% HBeAg-negative, 62% cirrhotics, 88% nor-al ALT, 74% undetectable HBV-DNA). Safety analysis focused on

lomerular (eGFR) and tubular renal function. Baseline was the startf TDF. During 5 years of TDF, 89 patients (28%) had to reduce TDFose after 31 months (2-75) (57 for eGFR decline, 30 for phosphateecline, 2 for both events). 11 additional patients (3.4%), who had totop TDF because of low blood phosphate levels, were successfullyescued by switching to ETV. 6 (2%) additional patients withdrewor gastrointestinal side effects. Overall, 106 patients (33%) eitherequired a dose reduction or withdrew from TDF for side effects,ith a 5-year estimated probability of 40%. None of the TDF patientseveloped acute renal failure or experienced a virological reboundfter dose adjustment. Virological response progressively increasedo 100% at year 5. 15 patients (5%) cleared HBsAg, 26 additionalatients had qHBsAg < 10 and 25 reached qHBsAg between 10 and00 IU/ml (Overall, 22% with qHBsAg < 100 IU/ml). HCC attack ratesere 1.3%/year in compensated cirrhotics and 0.2%/year in non cir-

hotics. No cases of clinical decompensation were recorded. Overall,patients (2%) were transplanted (all for HCC) and 16 (5%) died (7

or HCC, 7 for non liver causes and 2 for unknown reasons).Conclusions: To prevent glomerular and tubular damage,

roactive dose adjustments of TDF are necessary in a large pro-ortion of ADV-experienced, TDF treated CHB patients.


sease 47S (2015) e19–e42

T-46

TOTAL HEPATITIS B CORE ANTIGEN ANTIBODY, AQUANTITATIVE NON-INVASIVE MARKER OFHEPATITIS B VIRUS INDUCED LIVER DISEASE

F. Moriconi 2, Q. Yuan 1, L-W. Song 1,D. Cavallone 2, B. Cherubini 2, P. Colombatto 2,F. Oliveri 2, B. Coco 2, G. Ricco 2, F. Bonino 3,J.W-K. Shih 3, N-S. Xia 1, M.R. Brunetto 2

1 State Key Laboratory of Molecular Vaccinology andMolecular Diagnostics, National Institute ofDiagnostics and Vaccine Development in InfectiousDiseases, School of Public Health, Xiamen University,Xiamen, China2 Laboratory of Molecular Genetics and Pathology ofHepatitis Viruses, Hepatology Unit, Reference Centerof the Tuscany Region for Chronic Liver Disease andCancer, University Hospital of Pisa, Italy3 Digestive and Liver Disease, General Medicine IIUnit, University Hospital of Pisa, Italy

Background and Aim Non invasive immunologic markersof virus-induced-liver-disease are an unmet need. We com-pared the performances of quantitative total and IgM-anti-HBc inwell-characterized chronic-HBsAg-carriers. Methods. Sera (212)were obtained from 111 HBsAg-carriers followed-up for 40months (18-216 months) during different phases of chronic-HBV-genotype-D-infection; 10 HBeAg-positive, 25 inactive-carriers(HBV-DNA≤2000IU/ml, ALT < 30 U/L), 66 HBeAg-negative/anti-HBe-positive-CHB-patients and 10 with HDV-super-infection. In35 patients treated with Peg-IFN (180 �g/w for 12 m) sera wereobtained at baseline, end-of-therapy and week-24 off-therapyand in 22 treated with nucleos(t)ide-analogues (for 60 monthsmean, 42-134 months range) at baseline and end-of-follow-up.IgM- and total-anti-HBc were measured by Architect (Abbott, USA)and double-antigen-sandwich-immune (Wantai, China) assaysrespectively. Results. Total-anti-HBc was positive in all serawith lower levels (p < 0.001) in HBsAg-carriers without CHB(immune-tolerant, inactive and HDV-superinfected, mean 3.26,range 2.26-4.49 Log10 IU/ml) versus untreated-CHB (mean 4.68,range 2.76-5.54 Log10 IU/ml). Thirty of 212 (14.2%) sera wereIgM-anti-HBc-positive using acute-hepatitis-cut-off (1-S/CO value)and 102 (48.1%) with chronic-hepatitis-cut-off (0.130-S/CO). Total-and anti-HBc-IgM correlated (r2 = 0.4173). Total-anti-HBc declinedsignificantly in CHB-patients with sustained virological response(p < 0.001) treated with antivirals: groups a) inactive-carriers, b)baseline-untreated-HBeAg-negative-CHB, c) EOF-HBeAg-negative-CHB with SVR after Peg-IFN, d) EOF in NUC-treated-patients]; thelowest levels were found in SVR who cleared HBsAg subsequently.The distribution of total-anti-HBc and the differences betweengroups are clinically significant with values in responders to antivi-ral therapy (c and d) comparable to inactive carriers. IgM-anti-HBcvalues were in most of sera below the analytical-specificity cut-offof the assay even using the lower CHB cut-off of 0.130 S/CO. Duringspontaneous and therapy-induced CHB remissions and reactiva-tions both total- and IgM-anti-HBc correlated with ALT (p < 0.001,r = 0.351 and p = 0.008, r = 0.185 respectively). Conclusions. Total-anti-HBc qualifies as a useful marker of HBV-induced-liver-disease
that might help to discriminate major phases of chronic HBV infec-tion and to predict sustained response to antivirals.

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-47

OCEPREVIR OR TELAPREVIR PLUSEGINTERFERON/RIBAVIRIN IN HCV CHRONIC

NFECTION: THE REAL-LIFE EXPERIENCE OF THETALIAN ASSOCIATION OF HOSPITALEPATOLOGISTS (CLEO)

LEO DAAs Study Group, CLEO, Rome, Italy

Introduction: Boceprevir/Telaprevir (DAAs), approved foreimbursement in Italy in December 2012, were used from January013. Since then the group of the Association of Hospital Hepatol-gists (CLEO DAAs Study Group) has been deeply involved in usingAAs. In September 2013, the Governing Board of the Associationecided to collect data from Hospital Centers, where there werective members belonging to CLEO. For this reason, this study cane qualified as retrospective/prospective.

Aim: to check safety and efficacy of this type of treatment in theeal-world setting.

Patients and Methods: A database was prepared and used byll Centres for the data collection and updated continuously. Lastpdate: November 26, 2014. All patients consecutively treatedere included; data were analyzed according to the intention-to-

reat principle. HCV-RNA was analyzed using: COBAS TaqMan 2.0Roche) with LLQ 25IU/mL.

Results: 40 Centres enrolled 737 patients: male 64%; mediange 58 (range 18-78), of whom 20.2% over 65; mean BMI 25.6range 16-39); Genotype 1b (78.4%); fibrosis F3/4 (70%). DAAs used:elaprevir (67%); PEGIFN-2a (71%); patients naïve (28%), relapsers32%), non-responders (40%). Therapy was stopped in 13.4% casesecause of side-effects (anaemia 37%, rash 26%) or for virologicalailure (16.2%). Since the study is ongoing, we have 563 patientsho completed the follow-up. The RVR was achieved in 68% cases,

OT in 64%, while the SVR was achieved in 68% in F0-F1; 57% in2-F3 and 37% in F4. In cirrhotic aging > 65 the SVR was 34%. Thereere no fatalities.

Conclusions: DAAs, in everyday practice, are safe but with mod-rate efficacy. These data confirm the limited success of DAAs inertain groups of patients such as those widely represented in oureries: advanced fibrosis/cirrhosis, non-responders to PEGIFN/RIBAnd the over 65s. As for the SVR, the grade of fibrosis makes theifference.


-48

FFECTIVENESS OF ADALIBUMAB FOR PATIENTSITH PRIMARY SCLEROSING CHOLANGITIS

SSOCIATED WITH INFLAMMATORY BOWELISEASE

. Franceschet, N. Cazzagon, A. Floreani

Department of Surgery, Oncology andastroenterology, University of Padua, Padua, Italy

Background: Primary sclerosing cholangitis (PSC) is a chroniciliary disease with a marked comorbidity with extra-hepatic con-itions, mainly inflammatory bowel disease (IBD). The medicalreatment for PSC is still disappointing, whereas immunomodu-ators and biologics have been able to demonstrate efficacy in IBD.

Aim: To analyse: 1) the natural history of patients withSC ± associated with IBD; 2) the long-term efficacy of biologics
n patients with PSC and concomitant IBD.
Methods: 92 consecutive PSC patients (seen from 1987 to 2014)ere included in the study: 50 (54.3%) were males, and 42 (45.7%)

emales. The mean age at diagnosis was 32.0 ± 14.3 years, and

sease 47S (2015) e19–e42 e41

the mean follow-up was 103.8 ± 86 months. Forty-nine patients(53.3%) had associated IBD (38 ulcerative colitis, 10 Crohn’s disease,1 indeterminate colitis). Clinical activity and endoscopic severitywere scored according to the CD activity index and Mayo subscorefor endoscopy.

Results: Table 1 shows the major events occurred during thefollow-up:

Events PSC + IBD (N.49) PSC-IBD (N.43) p

OLTx 7 (14.3%) 7 (16.3%) 0.79CCA 0 2 (4.6%) 0.13HCC 2 (4.1%) 0 0.18Gallbladder cancer 1 (2.0%) 0 0.35Colo-rectal cancer 4 (8.2%) 0 0.05Death 5 (10.2%) 5 (11.6%) 0.83

Three patients with IBD experienced, as second-line treatment,Adalimumab (ADA), an anti-TNFa monoclonal antibody, previouswritten consensus. Patients were assessed before starting treat-ment, at month 6 and 12. ADA induction was 160 mg at week 0,and then 80 mg at week 2, while ADA maintenance treatment was40 mg every 2 weeks. After 6 and 12 months of ADA, a sustainedclinical remission of IBD was obtained; a reduction in ALT, GGT,alkaline phosphatase and Mayo PSC score was obtained.

Conclusions: This is the first study evaluating the efficacy of bio-logic agents in PSC. Promising results come from ADA for PSC + IBDduring a 12 months follow-up. Furthers studies are warranted toinvestigate the long-term tolerability and efficacy in such patients.


T-49

POSITIVE CORRELATION OF HIF2� ANDSERPINB3 IN HUMAN HEPATOCELLULARCARCINOMA: SELECTIVITY AND PROGNOSTICIMPLICATIONS

E. Morello 1, C. Turato 2, S. Cannito 1,M. Ruvoletto 2, S. Quarta 2, C. Paternostro 1,E. Novo 1, R. Autelli 1, S. Fasolato 2, I. Tusa 3,E. Rovida 3, S. Colombatto 4, A. Smedile 5,A. Vitale 6, G. Zanus 6, U. Cillo 6, M. Parola 1,P. Pontisso 2

1 Department Clinical and Biological Sciences, Unit ofExperimental Medicine, University of Turin, Turin,Italy2 Department Medicine, University of Padua, Padua,Italy3 Department Biomedical, Experimental and ClinicalSciences, University of Florence, Florence, Italy4 Department Oncology and5 Department Medical Sciences, University of Torino,Italy6 Department Surgical, Oncological andGastroenterological Sciences, University of Padua,Padua, Italy

Background SerpinB3, a cysteine-proteases inhibitor up-regulated in hepatocellular carcinoma (HCC), proposed asbiomarker of liver carcinogenesis and to stimulate epithelial-to-mesenchymal transition (EMT) and increased invasiveness in livercancer cells, has been suggested to be up-regulated by hypoxiathrough a HIF2�-dependent mechanism.

Aims To investigate the selectivity of HIF2�-related up-regulation of SerpinB3 and the in vivo prognostic relevance of thisrelationships.

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Methods Selectivity ofthe relationships between hypoxia,IF2� and SerpinB3 versus SerpinB4 polymorphic isoform expres-

ion has been investigated in control HepG2 cells or in HepG2 cellstably transfected to overexpress HIF1� or HIF2� by employingell and molecular biology techniques. HIF2� and SerpinB3 tran-cript levels were evaluated (quantitative real-time PCR) on frozenCC specimens obtained at the time of surgical resection from 67atients with cirrhosis of different aetiology. A series of 18 paraffin-

ncluded liver specimens from cirrhotic HCV patients carrying HCCas also included in the study for immuno-histochemistry analysis.

Results Selectivity of SerpinB3 up-regulation by hypoxia viaIF2� was established by the following experimental evidence:

) HIF2� (not HIF1�), binds to the Serpin B3 promoter (ChIPssay); ii) only HepG2 overexpressing HIF2� show SerpinB3 up-egulation; iii) hypoxic conditions do not affect expression oferpinB4. Immuno-histochemistry and transcript analysis, per-ormed in human HCC specimens, revealed co-localization of thewo proteins in liver cancer cells and the existence of a positiveorrelation (Spearman r coefficient 0,3533, p < 0.01) between HIF-� and SERPINB3 transcript levels. Moreover, the highest levels ofIF-2� transcripts were found in specimens with the highest levelsf SerpinB3 transcripts obtained from the most aggressive sub-et of HCC cases, correlating with the highest rate of early tumourecurrence.

Conclusions A positive correlation between HIF2� and Ser-inB3 expression exists in human HCC specimens which is selectiveor the SerpinB3 isoform and has prognostic implications.


-50

HE ACTIVATION OF NF-KB, PREGNANE XECEPTOR, AND CONSTITUTIVE ANDROSTANEECEPTOR IS MODULATED BY THE DEGREE OFHOLESTASIS

. Gabbia 1, T. Baldovin 2, R. Lazzari 2, C. Mescoli 3,. Albertoni 3, V. Baldo 2, A. Floreani 4,. De Martin 1

Department of Pharmaceutical andharmacological Sciences, University of Padua,adua, ItalyDepartment of Molecular Medicine, University ofadua, Padua, ItalyDepartment of Medicine, University of Padua,adua, ItalyDepartment of Surgery, Oncology and
astroenterology, University of Padua, Padua, Italy
Introduction: NF-kB is known to be activated in the early stagesf cholestasis, acting a reduction of liver injury. Pregnane X Recep-

sease 47S (2015) e19–e42

tor (PXR) and Constitutive Androstane Receptor (CAR) are twonuclear receptors (NRs) that regulate bile acid metabolism andtransport. A mutual negative crosstalk between NF-kB and NRs hasbeen reported, but conflicting data are available on the impact oftheir relationship in cholestasis.

Aim: To analyse the changes in activation of NF-kB, PXR, andCAR in an experimental model of cholestasis.

Methods: Cholestasis was induced in 8 male Wistar rats bybile duct ligation; 4 sham-operated rats were used as controls. Thedegree of cholestasis was defined on the basis of histologic exam-ination of liver sections and serum concentration of albumin, ALT,GGT, bilirubin. Activation of PXR, CAR and NF-kB was evaluated byWestern blot analysis on nuclear liver fractions.

Results: A 2-fold increase in activation of NF-kB was observed inearly stage of cholestasis (p < 0.05 with respect to Sham), whereasthe nuclear translocation of NF-kB was completely abolished inthe late stage (p < 0.001). A significant increase in PXR activationwas observed in late stage of cholestasis compared with both earlystages and controls (2- and 1.5-fold, p < 0.01 and 0.05, respectively).CAR nuclear expression was 3.4-fold higher in early cholestatic ratsthan in controls (p < 0.001), whereas CAR activation was virtuallyabolished when cholestasis became severe (p < 0.05 and p < 0.001vs. sham and early-stage cholestatic rats, respectively).

Conclusions: We provided new evidences of the strict rela-tionship between NRs and NF-kB activation, suggesting a differenteffect of NF-kB on PXR and CAR activation, which is dramaticallyenhanced in severe cholestasis. Therefore, a different modulationof the NF-kB pathway by acting on PXR and CAR according to the

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-01

TILITY BASED CRITERIA TO SELECT PATIENTSITH HEPATOCELLULAR CARCINOMA FOR

IVER TRANSPLANTATION: A MULTICENTREOHORT STUDY

. Vitale 1, F. Farinati 2, P. Burra 2, F. Trevisani 3,.G. Giannini 4, G. Spolverato 1,. Cillo 1, for the Italian Liver Cancer (ITA.LI.CA) group

Dipartimento di Chirurgia Generale e Trapianto’Organo, Unità di Chirurgia Epatobiliare e Trapiantipatici, Università di Padova, Padova, ItalyDivisione di Gastroenterologia, Azienda Universitài Padova, Padova, ItalyDipartimento di Scienze Mediche e Chirurgiche,nità di Semeiotica Medica, Alma Matertudiorum–Università di Bologna, Bologna, ItalyDipartimento di Medicina Interna, Unità diastroenterologia, Università di Genova, Genova,enova, Italy

Background. The lifetime utility of liver transplantation (LT) inatients with hepatocellular carcinoma is still controversial. Theim of this study was to understand when LT is cost-effective forCC patients in order to suggest new potential transplant selectionriteria.

Methods. The overall design of this study involved a real cohortf transplantable Italian HCC patients (n = 2419 selected from theTA.LI.CA database) undergoing non-transplant therapies for whomsurvival analysis and calculation of direct costs of therapies wereerformed, and then a Markov model to calculate the survival ben-fit and cost-utility of LT over non-LT therapies. Post LT survival wasalculated using the �-fetoprotein (AFP) model based on AFP valuesnd radiological size and number of nodules. Primary endpoint waset Health Benefit (NHB), defined as LT survival benefit in qualitydjusted life years (QALYs) minus incremental costs ($)/willing-ess to pay ($32,946, corresponding to the Italian Gross Domesticroduct).

Results. We developed a prognostically accurate survival modelredicting non-LT survival, based on AFP model variables, Child
ugh classes, and type of alternative therapy (resection, loco-egional, or best supportive care). The calculated median cost ofon-LT therapies per patient was $21,670. The Monte Carlo simu-
ation showed that in patients with resectable child A HCC, the NHB

590-8658/$36.00

of LT was always negative, independently from the AFP model val-ues, while in unresectable HCC patients, the NHB of LT was positivefor AFP model values ≤ 4, and negative for values > 4.

Conclusion. LT proved to be cost-effective in HCC patients withunresectable tumor and AFP model values ≤ 4.


F-02

STIMULATION OF NUCLEAR RECEPTOR PPAR-�LIMITS NF-KB-DEPENDENT INFLAMMATION INCYSTIC FIBROSIS BILIARY EPITHELIUM

R. Scirpo 1,2, R. Fiorotto 1, A. Villani 1, L. Fabris 3,M. Strazzabosco 1,2

1 Department of Internal Medicine, Yale University,New Haven, USA2 Department of Surgery and Traslational Medicine,University of Milano-Bicocca, Monza, Italy3 Department of Surgical Oncological andGastroenterological Science, University of Padua,Padua, Italy

Background. Cystic Fibrosis-associated liver disease (CFLD) isa chronic cholangiopathy that negatively affects life of CysticFibrosis patients. We have recently shown that altered TLR/NF-kB-dependent biliary innate immune mechanisms contribute tothe pathogenesis of CFLD, and may represent novel therapeutictargets for CFLD. Nuclear receptors (NRs) are ligand-dependenttranscription factors that, by controlling gene expression, regulateseveral intracellular functions. Selected classes of NRs, includ-ing peroxisome proliferator-activated receptor � (PPAR-�), alsocounter-regulate inflammation in several tissues, by cross-talkingwith TLR-dependent signaling pathways. We hypothesized thatpharmacologic activation of nuclear receptor PPAR-� would inhibitthe TLR/NF-kB-dependent inflammation in Cftr-defective biliaryepithelium.

Results. The gene and protein expression of PPAR-� is increasedin CF cholangiocytes respect to WT cells, but does not correlatewith a higher expression of PPAR-� target genes (Acaa1b, Angptl4
and Hmgcs2), indicating that the receptor is not more active. Con-sistently, Cftr-KO cells show a decreased production of PPAR-�endogenous ligands that might limit a proper activation of thereceptor. Treatment with LPS caused a higher NF-kB activation and

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F-kB-dependent cytokine secretion in Cftr-KO cells, compared toontrols. In the presence of PPAR-� agonists, the activation of NF-kBnd the secretion of proinflammatory cytokines LIX, MCP-1, MIP-and G-CSF were significantly inhibited at baseline and after LPS

hallenge. Modulation of NF-kB-dependent inflammation by PPAR-resulted from the induction of the NF-kB negative regulator IkB�.

inally, stimulation of PPAR-� in vivo significantly attenuated bil-ary damage and inflammation in Cftr-KO mice after DSS-inducedortal endotoxemia.

Conclusions: Our studies unravel a novel mechanism of reg-lation of biliary epithelium innate immunity and suggest thattimulation of PPAR-� signaling by synthetic agonists may rep-esent a valuable therapeutic option to limit biliary inflammationn CFLD. We expect that they might be applicable to other organsffected by CF and to the treatment of other inflammatory cholan-iopathies.


-03

EEP-SEQUENCING ANALYSIS DEMONSTRATESHE PERSISTENCE OF PRE-TRANSPLANT HCVOMINANT VARIANTS WITHIN A MOREOMOGENEOUS QUASISPECIES AFTER LIVERRANSPLANTATION (LT) IN CHOLESTATICEPATITIS C PATIENTS

. Gambato 1,2, J. Gregori 3,4, J. Quer 3,. Caro-Pérez 1, G. Crespo 1, M. Navasa 1,. Pérez-del-Pulgar 1, X. Forns 1

Liver Unit, Hospital Clínic, IDIBAPS, Ciberehd,arcelona, SpainMultivisceral Transplant Unit, Department ofurgery, Oncology and Gastroenterology, Paduaniversity Hospital, Padua, ItalyLiver Unit, Vall d’Hebron Institut deecerca-Hospital Universitari Vall d’Hebron,IBERehd, Barcelona, SpainRoche Diagnostics SL. Sant Cugat del Vallès,arcelona, Spain

Introduction. Little is known about the pathogenic mechanismsmplicated in cholestatic hepatitis C (CH). Host failure to mountpecific T-cell responses against HCV has been implicated in theathogenesis. However, a direct role of HCV is not well established.

Aim. The aim of this study was to analyze, for the first time,he HCV quasispecies evolution in patients with CH (compared tocontrol group) by ultra-deep pyrosequencing (UDPS).

Materials and Methods. 13 HCV-infected liver transplant recip-ents with CH and 10 with mild recurrence (control group) haveeen analyzed so far. A serum sample at the time of LT and a sec-nd sample obtained 1-3 months after LT were analyzed for eachatient. UDPS of a NS5B fragment (339 nucleotides) was performedsing the 454 GS Junior platform.

Results. Donor age (p = 0.007), recipient age (p = 0.022) and viraload after LT (p < 0.001) were significantly higher in patients withH compared to controls. The mean number of reads per sam-le was 6146 (range 2491-25820). NS5B quasispecies complexityefore LT was similar in CH and control group (p = 0.396). On theontrary, genetic diversity and mutation frequency of NS5B quasis-ecies decreased significantly after LT in patients with CH (p = 0.013nd p = 0.013), but not in the control group (p = 0.069 and p = 0.327).
nterestingly, phylogenetic analysis showed that the major HCVariant pre-LT successfully propagated and remained as the major
sease 47S (2015) e43–e66

variant after LT in 64% of patients with CH, but only in 12% of con-trols (p = 0.026).

Conclusions. A marked homogenization of HCV quasispecieswas demonstrated in HCV-infected patients who developed CHafter LT. In addition, in most CH patients, the major HCV strainbefore LT remained as dominant after LT. The latter suggests that, inthe context of immunosuppresion, the propagation of more fittedviral strains could play a role in the pathogenesis of CH.


F-04

OVERALL SURVIVAL IN INTERMEDIATE-STAGEHEPATOCELLULAR CARCINOMA (HCC) PATIENTSAFTER FIRST TRANSARTERIALCHEMOEMBOLIZATION (TACE): PROPOSAL OF ANEW SCORING SYSTEM

R. Sacco 1, B. Ginanni 3, V. Mismas 1, G. Masi 2,P. De Simone 2, A. Romano 1, G. Bresci 1,C. Bartolozzi 3, I. Bargellini 3

1 Department of Gastroenterology, Pisa UniversityHospital, Pisa, Italy2 Department of Oncology, Pisa University Hospital,Pisa, Italy3 Department of Radiology, Pisa University Hospital,Pisa, Italy

Introduction TACE is the standard treatment for patients withintermediate-stage HCC. However, prognostic factors for survivalafter the first TACE cycle remain unclear.

Aims We correlated pre-treatment characteristics and responseto therapy with overall survival (OS) and time to progression (TTP),in order to propose a scoring system aimed at facilitating clinicaldecision after the first TACE.

Patients and methods We retrospectively analyzed 149 patientswith intermediate-stage HCC who received ≥1 TACE cycle(s).Univariate and multivariate analysis were used to correlate pre-treatment characteristics and response to TACE with OS and TTP.Predictive factors were used to define a score for each patient.

Results Median OS was 23 (95% CI 11.5-27) months, and medianTTP was 11 months (7-11). Complete response was reported in 63patients (42.3%) and partial response (PR) in 71 (47.7%).

Age >65 years (HR 1.77; 95% CI: 1.18-2.67), ascites (HR 2.44; 95%CI 1.32-4.29), total diameter of nodules >61 mm (HR: 1.96; 95% CI1.28-3.08) and response at 1 month (HR 1.70; 95% CI 1.30-2.20)were predictors of survival and were used to build the scoring sys-tem. Age >65 years, ascites, total diameter of nodules >1 accountedfor one score point; PR, stable disease and progression of diseaseaccounted for 1, 2 and 3 score points, respectively. The score pointswere summed to calculate a single score for each patient.

Patients with score 0-1 had a longer OS (57.8 months) and TTP(12.7 months) than those with score 2-3 (21.1 and 8.2 months) orscore 4-6 (8.0 and 6.3 months) (p < 0.001 for both comparisons).

Conclusions This scoring system may allow the identification ofthree groups of patients with different prognosis after a first TACEcycle and may therefore help guide clinical decisions, in particular

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-05

REATMENT OF ADVANCED HEPATOCELLULARARCINOMA (HCC): A SINGLE CENTRE COSTNALYSIS OF YTTRIUM 90 TRANS-ARTERIALADIO-EMBOLIZATION (TARE) VERSUSORAFENIB

.G. Lucà 1, M. De Giorgio 1, G. Magini 1,. Tortora 4, L. Sangiovanni 1, G. Gaffuri 1,. Baldan 1, G. Virotta 3, R. Nani 2, S. Fagiuoli 1

Department of Internal Medicine, Papa GiovanniXIII Hospital, Bergamo, ItalyDepartment of Radiology, Papa Giovanni XXIIIospital, Bergamo, ItalyDepartment of Nuclear Medicine, Papa GiovanniXIII Hospital, Bergamo, ItalyDivision of Internal Medicine and Gatroenterolgy,epartment of Internal medicine, A. Gemelliospital, Rome, Italy

INTRODUCTION: In a selected population of advanced HCCatients TARE seems to give promising results.

AIM: To compare a single Centre costs for Sorafenib and TAREMETHODS: 166 consecutive advanced HCC patients (M 86%,

edian age 67 yrs) treated with Sorafenib between June 2009 andune 2014 (Group SOR) and 19 (M 84%, median age 64 yrs, BCLC B1,5%) treated with TARE between June 2011 and June 2014 (GroupARE) were compared.

Patients were grouped for treatment time (Group SOR 1: < 2onths of treatment; Group SOR 2: > 2 months of treatment).roup SOR 1 drug expenses were not further considered accord-

ng with the agreed “pay for result policy”. In group SOR 2, 24 ptsgroup SOR 3) presented monolobar disease and no methastasisBCLC B 54%) so as to be potentially treated with TARE. Sorafenibost was calculated according with the total effective caps intakeor each patients (28,7 Euro/caps).

TARE costs included Yttrium-90 and hospitalization expense17.761 Euro/patient).

RESULTS: Median time of treatment in Group SOR 3 was 272ays (154-994) with a median intake of 2,8 tablets/day. All theatients in the group TARE were treated with a single treatmentession. Median follow up time was respectively 476 days (154-518) for the Group SOR 3 and 266 days (47-867) for the GroupARE.

Survivals at 12, 18, 24 and 36 months were respectively 66,7%,7,5%, 24,3% and 19.4% for the Group SOR 3 and 64.1%, 64.1%, 64.1%nd 32% for the Group TARE (Log Rank p = 0.446). The GLOBAL treat-

sease 47S (2015) e43–e66 e45

ment costs for Group SOR 3 and for Group TARE were D 671.809,6and D 337.459,00 respectively (median D 27992/pt in SOR3 vs D17761/pt in TARE; p = 0.028).

CONCLUSION: TARE treatment in advanced HCC may reducecosts with survivals not significantly different from Sorafenib.


F-06

CORONARY MICROVASCULAR DYSFUNCTION INPATIENTS WITH PRIMARY BILIARY CIRRHOSISWITHOUT METABOLIC SYNDROME: A HINT FORTHEIR INCREASED CARDIOVASCULAR RISK

N. Cazzagon 1, C. Dal Lin 2, G. Famoso 2,I. Franceschet 1, A. Floreani 1, F. Tona 2

1 Department of Surgery, Oncology andGastroenterology, University of Padua, Padua, Italy2 Department of Cardiac, Thoracic and VascularSciences, University of Padua, Padua, Italy

Background: Primary biliary cirrhosis (PBC) is characterizedby a long natural history and a low incidence of cardiovascularevents despite high serum cholesterol levels. Metabolic syndromemay increase the cardiovascular risk, however. Coronary flowreserve (CFR) is widely used to examine the integrity of coronarymicrovascular circulation and it is well recognized as a predictor ofcardiovascular outcome.

Aim: To evaluate the risk of coronary microvascular dysfunctionin patients with PBC without metabolic syndrome.

Methods: 29 PBC patients (27 F, aged 56 ± 11 years) withoutclinical evidence of heart disease and without metabolic syn-drome, and 29 sex- and age-matched healthy controls underwentCFR by transthoracic Doppler echocardiography (TDE). Coronaryflow velocity in the left anterior descending coronary artery wasdetected by TDE at rest and during iv. adenosine infusion. CFR wasthe ratio of hyperaemic diastolic flow velocity (DFV) to resting DFV.

Results: The median time between the onset of symptoms andCFR assessment was 7 years (interquartile range 3-10.5 years). InPBC patients CFR was significantly lower than in controls (3.1 ± 0.3vs 3.8 ± 0.4, p < 0.0002) (Fig. A). CFR was inversely related to thetime from diagnosis (r = -0.389, p = 0.03) (Fig. B) and age (r = -0.370,p = 0.04), and positively correlated with TSH (r = 0.411, p = 0.03). Norelationship was found between CFR and LDL-cholesterol.

Conclusions: CFR is impaired in PBC patients and is correlatedwith the length of disease, suggesting a negative effect of PBC oncoronary microcirculation that may contribute to the increased car-diovascular risk, even in patients without metabolic syndrome.


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-07

HE TM6SF2 E167 K VARIANT IS ANNDEPENDENT PREDICTOR OF SEVERE LIVERTEATOSIS IN CHRONIC HEPATITIS C

. Coppola 1, R. Zampino 2, G. Cirillo 3,. Stanzione 4, M. Macera 1, A. Boemio 2,

. Grandone 3, M. Pisaturo 1, A. Marrone 2,.E. Adinolfi 2, E. Sagnelli 1,. Miraglia del Giudice 3

Department of Mental Health and Public Medicine,ection of Infectious Diseases, Second University ofaples, Naples, ITALYInternal Medicine and Hepatology, Secondniversity of Naples, Naples, ITALYDepartment of Pediatrics, Second University ofaples, Naples, ITALYDepartment of Clinical and Experimental Medicinend Surgery, Second University of Naples, Naples,TALY

Background: A common non-synonymous polymorphism,167 K, in transmembrane 6 superfamily member 2 (TM6SF2) geneas been recently associated with an increased hepatic triglycerideontent, dyslipidemia and liver fibrosis in NAFLD patients.

Aim: We investigated on a possible associations betweenM6SF2 variants and degree of liver lesions in chronic hepatitis.

Methods: 148 consecutive patients with biopsy proven anti-CV/HCV-RNA-positive chronic hepatitis, naive for antiviral

herapy, were genotyped for TM6SF2 E167 K and PNAPL3 I148 Mariants by a TaqMan Assay. These patients aged 50.9 ± 12.8, 54.7%ere males, 14.9% with a history of alcohol abuse, 62.2% with HCV

enotype 1 and 9.3% with genotype 3.Results: 130 (87.8%) patients were TM6SF2 167E homozygotes

nd 18 (12.2%) heterozygotes for the E167 K variant. The preva-ences of patients with the PNAPL3 I/I, I/M or M/M were 26.3%,9.1% and 14.6%, respectively. Patients with different TM6SF2ariants did not show significant differences in demographics, bio-hemical data including cholesterol and triglyceride, HCV load,revalence of patients with HCV genotype 3 and distribution ofNPLA3 variants. As compared with the 130 homozygotes forM6SF2 167E allele, the 18 patients with TM6SF2 E167 K vari-nt showed a higher degree (scale from 1 to 4) of liver steatosis1.9 ± 1.3 vs. 1.1 ± 1.1, p = 0.02) and a higher prevalence of casesith a steatosis score ≥ 3 (33.3% vs. 12.3%, p = 0.02); no difference

n necroinflammation nor in fibrosis scores was observed. A gen-ral linear model identified as independent predictors of steatosishe TM6SF2 E167 K, the PNAPL3 M148 M variants and the waistircumference (p = 0.0376, p = 0.0069 and p = 0.0273, respectively).he effect of the PNAPL3 M/M variant on liver steatosis looks sig-ificantly enhanced when associated with the TM6SF2 E/K variantOR 11.4, CI 95% 1.06-134, p = 0.04).

Conclusions: This study is the first demonstration that TM6SF2
167 K variant is an independent predictor of severe liver steatosisn chronic hepatitis C.

sease 47S (2015) e43–e66

F-08

METABOLIC SYNDROME AFTER LIVERTRANSPLANTATION: HAVE RISK FACTORSCHANGED OVER TIME?

E. Vanni 1, R. Ibrahim Kamal Jouness 1,S. Mirabella 2, A. Marengo 1, A. Milan 3, C. Rosso 1,V. Boano 1, E. Mosso 1, C. Di Stefano 3, E. Nada 2,M. Rizzetto 1, M. Salizzoni 2, R. Romagnoli 2,E. Bugianesi 1

1 Department of Medical Sciences, Division ofGastroenterology, University of Turin, Turin, Italy2 Liver Transplant Center, University of Turin, Turin,Italy3 Department of Medical Sciences, Division ofInternal Medicine, University of Turin, Turin, Italy

Background and Aims: Post-Transplant Metabolic Syndrome(PTMS) is common among liver transplantation (OLT) recipientsThe aim of this study is the definition of the factors contribut-ing to PTMS development with particular regard to changes in theimmunosuppressive regimens.

Methods: Three hundred and ninety-three patients who under-went OLT at Turin Liver Transplant Center were prospectivelyenrolled (group A: pre-mTOR inhibitors - OLT 1998-2003, n = 167;group B: m-TOR inhibitors - OLT 2008-2012, n = 226). Overall themedian follow-up was 60 months (1-204), 144 (83-204) in group Aand 48 months (1-60) in group B. Clinical, laboratory parameters,prescribed medications, were recorded every 6-12 months. Clinicalfeatures of the donors and histological data of the allograft were col-lected. Multivariate analysis were performed to identify risk factorsassociated with PTMS development.

Results: Clinical features and OLT indications were compara-ble between the two groups. Cryptogenic cirrhosis was an OLTindication in 2.8% (11/393) without temporal changes. Overall pre-OLT MS was found in 1.8% (7/393) subjects, without temporalchanges, while PTMS in 24.9% (98/393; group A 37.9%, group B22.6%, p = 0.003). Overall multivariate analysis identified pre-OLThyperglycemia and post-OLT weight gain as predictive risk fac-tors for PTMS. However, independent risk factors were differentin the two groups: post-OLT weight gain (OR 1.53, 95% CI 1.26-1.85, p = 0.0001) and post-OLT US liver steatosis (OR 4.12, 95% CI1.48-11.4, p = 0.006) in group A; post-OLT weight gain (OR 1.36, 95%CI 1.17-1.57, p = 0.0001), pre-OLT hyperglycemia (OR 3.57, 95% CI1.28-9.88, p = 0.014), macrovacuolar steatosis ≥20% in the allograft(OR 4.74, 95% CI 1.23-18.27, p = 0.024) and treatment with m-TORinhibitors (OR 17.43, 95% CI 3.36-90.39, p = 0.001) in group B.

Conclusions: Overall, host-related factors contribute to PTMSdevelopment, but the introduction of mTOR inhibitors is an

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-09

LINICAL EVALUATION OF CIRCULATINGICRORNAS AS POTENTIAL BIOMARKERS OFEPATOCELLULAR CARCINOMA IN PATIENTSITH CHRONIC HBV INFECTION

.P. Caviglia 1, M.L. Abate 1, E. Petrini 2, S. Gaia 2,. Manzini 3, P. Carucci 2, M. Rizzetto 1,2,. Smedile 1,2

Department of Medical Sciences, University ofurin, Turin, ItalyDepartment of Gastroenterology and Hepatology,ittà della Salute e della Scienza Hospital, Turin, ItalyBlood Bank, Città della Salute e della Scienzaospital, Turin, Italy

Introduction. Several studies showed that aberrant miRNAxpression is associated with development and progression ofepatocellular carcinoma (HCC).

Aim. To examine whether some commonly deregulated miRNAsmiR-122, miR-21, miR-221 and miR-16) in HBV-related HCC mayerve as diagnostic markers.

Materials and Methods. Serum expression of miR-122, miR-1, miR-221 and miR-16 was evaluated by real-time quantitativeT-PCR in 33 patients with HBV-related HCC (61 ± 10 years;/M = 4/29), 30 patients with HBV-related cirrhosis (53 ± 11 years;/M = 11/19) and 27 blood donors as healthy controls (54 ± 8 years;/M = 9/18).

Results. Median levels of miR-16, miR-122 and miR-221 wereignificantly different in patients with HCC or cirrhosis compared toealthy controls (p < 0.001) whereas, only miR-122 levels differed

n patients with HCC from cirrhotic patients (p = 0.024). miR-122evels were significantly higher in patients with multifocal HCChan in patients with a single lesion (p = 0.024). Expression levelsf mir-21 were similar in the 3 groups. Area under the curve (AUC)nalysis showed that serum levels of miR-122, miR-122 + miR-21, miR-122 + miR-16 and miR-122 + miR-221 + miR-16, are ableo differentiate patients with HCC from patients with cirrhosisAUC = 0.675; AUC = 0.704; AUC = 0.681; AUC = 0.703, respectively).

oreover, miR-16, miR-122 and miR-221, alone or in combina-ion, were able to discriminate patients with HCC or cirrhosis fromealthy controls (AUC > 0.9). In addition, a correlation betweeniR-122 levels and HCC nodules number (R = 0.390; p = 0.036), and

etween miR-16 and miR-122 levels, and ALT values (R = -0.464,= 0.034; R = 0.449, p = 0.536, respectively) was found.

Conclusions. Among the four microRNAs analyzed, only miR-22 significantly discriminates patients with HCC from cirrhoticatients and patients with HCC or cirrhosis from controls. miR-22 appears to reflect liver necro-inflammation, since we observedpositive correlation with ALT levels. Moreover, the correlation

etween miR-122 expression levels and HCC with multi-nodules,uggests the possible use of this miRNA for tumor stadiation. Nev-
rtheless, miR-122 AUC values were not sufficiently high for HCCcreening purposes.

sease 47S (2015) e43–e66 e47

F-10

THE TWO THIENO-TRIAZOLODIAZEPINES WEB2086 AND WEB 2170 BLOCKHEPATOCARCINOMA PROGRESSION BYHSP-DEPENDENT CLIENT KINOMA PROTEINUBIQUITINATION

E. Ceni 1, C. Malentacchi 1, M. Tarocchi 1,G. Marroncini 2, S. Polvani 1, T. Mello 1,S. Tempesti 1, A. Galli 1,2

1 Experimental and Clinical Biomedical SciencesMario Serio, University of Florence, Florence, Italy2 FiorGen Foundation, Florence, Italy

Introduction: L’HCC is one of the most common malignanciesworldwide. So far there is no effective chemotherapeutic treat-ment for HCC and the prognosis of advance stage remains poor.The thieno-triazolodiazepine are well known anti-inflammatorydrugs that acts as PAF-receptor antagonists. Recently an anti-neoplastic pleiotropic effect of this molecules have been shown,where treatment with thieno-triazolodiazepine (WEB2086 andWEB2170) induce differentiation, growth arrest and apoptosis inmurine and human leukemia cells. The effects of WEBs on HCCremain untested. Aim of this study was to investigate the anti-tumor efficacy of the WEBs in vitro and in vivo models of HCC.

Material and Methods Results: WEBs were able to reduce pro-liferation of cancer cell lines (HepG2, Hep3B, HuH7 and Hepa 1-6)as assessed by thymidine incorporation and eliminates the abil-ity of these cells to form colonies in soft agar that was associatedwith cell cycle arrest, apoptosis, and senescence. In addition, WEBsimpair hepatoma cell migration in a Wound Healing assay andchemoinvasion on Matrigel. WEBs administration in HBV trans-genic mice reduces the number and the dimension of tumor masses.Antitumorigenic effects in mice were also confirmed in a HepG2xenograft model of HCC. A 2D-DIGE proteome study highlights thatthe WEB2086 inhibits the binding processes in particular inhibitsprotein binding of ATP and GTP and the abrogation of this cellularfunction results kinoma inhibition. In addition 2D-DIGE analy-sis showed that WEBs down-regulate chaperone proteins such ashsp90 and hsp70 that are involved in protein refolding, an impor-tant mechanism in tumor cell resistance. The down-regulationdell’Hsp90 lead to failure of “clients” protein refolding and mediatetheir degradation.

Conclusions: The thieno-triazolodiazepines are able to reduceHCC progression in human and murine HCC models blocking cancer

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-11

LINICAL RELEVANCE OF NEXT GENERATIONEQUENCING ON BASELINE DETECTION OFINORITY RESISTANCE ASSOCIATED VARIANTS

N HCV-1 PATIENTS TREATED WITH PROTEASENHIBITORS

. Armenia 1, L. Carioti 1, V.C. Di Maio 1,.C. Bellocchi 1, D. Di Paolo 2, F. Guerrieri 3,

. Calvo 4, V. Cento 1, M. Tontodonati 5,. Micheli 6, F. De Leonardis 2, M. Aragri 1,. Polilli 7, A. Manunta 8, C. Magni 6,.P. Antonucci 1, F. De Luca 1, C. Sarrecchia 9,. Bertoli 1, I. Lenci 2, S. Francioso 2,.M. Santoro 1, J. Vecchiet 5, S. Marenco 10,

. Picciotto 10, L. Nosotti 11, F. Morisco 12,. Bruno 13, M. Puoti 14, S. Babudieri 8, M.S. Mura 8,. Andreoni 9, G. Rizzardini 6, G. Parruti 7,. Levrero 4, M. Angelico 2, C.F. Perno 1,

. Ceccherini-Silberstein 1

Department of Experimental Medicine and Surgery,niversity of Rome “Tor Vergata”, Rome, ItalyHepatology Unit, University Hospital of Rome “Torergata”, Rome, ItalyCenter for Life Nano Science, CNLS@SAPIENZA, ITLa Sapienza” University, Rome, ItalyInfectious Disease Clinic, Chieti, ItalyHospital Sacco of Milan, Milan, ItalyInfectious Disease Unit, Pescara General Hospital,escara, ItalyDepartment of Clinical and Experimental Medicine,niversity of Sassari, Sassari, ItalyInfectious Disease, University Hospital of Rome “Torergata”, Rome, Italy0 S. Martino University Hospital, Genoa, Italy1 Hepatology Unit, National Institute of Health,igration and Poverty, Rome, Italy

2 University “Federico II”, Naples, Italy3 Internal Medicine, Gastroenterology andepatology, Azienda Ospedaliera Fatebenefratelli eftalmico, Milan, Italy

4 Hospital Niguarda Ca’Granda, Milan, Italy

Background and Aims: This study aims to evaluate thelinical relevance of next-generation-sequencing on the base-ine detection of minority resistance-associated-variants (RAVs)n chronic HCV-1 infected patients treated with telaprevir/oceprevir + pegIFN/ribavirin.

Methods: NS3-protease sequences of 25 selected patients18 virological-failures and 7 responders) treated with telapre-ir (N = 20) or boceprevir (N = 5) + pegIFN/ribavirin were analyzed.etection of NS3-RAVs (V36ALM/T54AS/V55A/Q80 K/R155KT/156STV/V170A) was performed by Sanger-sequencing, 454-

unior-pyrosequencing (UDPS) and MiSeq-illumina (MiSeq).UDPS sequences were analyzed by a home-made-pipeline

>3,000 sequences/patient; cut-off > 0.1%); Miseq sequences werenalyzed by VirVarSeq (>40,000 sequences/patient; cut-off ≥ 0.3%).

Results: At baseline, 5/25 patients (20%) presented NS3-RAVsy Sanger, UDPS and MiSeq (Q80K = 3; T54S = 1; V36L + Q80K = 1,revalence >98%). Of them, 4 patients experienced virological-ailure and 1 (with Q80 K) achieved sustained-virological-response
SVR). Additional baseline minority-RAVs (Q80 K/V170A) wereound by both UDPS and MiSeq in 2 patients, all with preva-ence < 2.6%. However, their presence was not directly associated
ith virological-failure. Indeed, 1 previous-null-responder with

sease 47S (2015) e43–e66

baseline minority Q80 K failed telaprevir-triple-therapy with dif-ferent RAVs (V36AM + R155K + T156ST). The other patient, previouspartial-responder, with baseline minority V170A achieved SVR.

By using MiSeq, analyzing a larger number of sequences(median[IQR]:843,492[342,499-1,006,108]), a high number of NS3-RAVs (N = 43,cut-off ≥0.3%) were detected in 19/25 patients (76%).In particular, V55A was the most prevalent RAV detected (52%),followed by V36A (36%), Q80 K (28%), and T54A (8%). The presenceof these mutations at baseline was similar between respondersand virological-failures. Interestingly, a unique HCV-1a infectedpatient previous-non-responder with 10 baseline minority-RAVs (V36A:0.41%; T54S:0.33%; V55A:0.47%, Q80 K:0.36%;R155 K:0.55%; R155 T:0.59%; A156S:1.19%; A156 T:0.64%;A156 V:0.50%, V170A:0.57%, with a median[IQR] mutational-loadof 19,567[15,870-21,987] IU/ml) failed telaprevir-triple-therapy,with A156 T (week-2) and later with V36M + R155 K (week-10).

Conclusions: Both UDPS and MiSeq detected baseline RAVs atvery low frequency. However, these variants were not necessarilydetected at virological-failure. Further investigations are needed toclarify the clinical relevance of minority RAVs at frequency below1%.


F-12

CHRONIC INTERMITTENT HYPOXIA ISASSOCIATED WITH LIVER DAMAGE ANDATHEROSCLEROSIS IN PATIENTS WITHNON-ALCOHOLIC FATTY LIVER DISEASE

S. Petta 1, O. Marrone 2, D. Torres 3,M. Buttacavoli 4, C. Cammà 1, V. Di Marco 1,A. Licata 1, A. Lo Bue 2, G. Parrinello 3, A. Pinto 3,A. Selvaggio 2, A. Tuttolomondo 3, A. Craxì 1,M. Bonsignore 4

1 Sezione di Gastroenterologia, DiBiMIS, University ofPalermo, Palermo, Italy2 National Research Council, Institute of Biomedicineand Molecular Immunology, Palermo, Italy3 Sezione di Medicina Interna, DiBiMIS,University ofPalermo, Palermo, Italy4 Sezione di Pneumologia, DiBiMIS,University ofPalermo, Palermo, Italy

Background: Obstructive sleep apnea syndrome(OSAS) hasbeen reported as a new risk factor for metabolic disturbances,including cardiovascular alterations. Growing data are also avail-able among bariatric nonalcoholic fatty liver disease(NAFLD)patients on the impact of obstructive sleep apnea syndrome OSASon liver damage in NAFLD. We assessed whether OSAS is associatedwith severity of liver fibrosis and carotid atherosclerosis in NAFLDpatients without morbid obesity.

Methods: 126 consecutive biopsy-proven(Kleiner score) NAFLDpatients assessed for anthropometric, biochemical, and metabolicfeatures underwent ultrasonographic carotid assessment and STOPBANG questionnaire for estimate of low or high OSAS risk. A carotidplaque was defined as a focal thickening > 1.3 mm at the level ofeither common and internal carotid arteries or bifurcations. 50patients accepted to perform nocturnal cardiorespiratory polygra-phy, and OSAS was defined if the as an apnea/hypopnea index AHIindex was≥5.

Results: The prevalence of high OSAS risk was similar in patients
without and with polygraphy(76% vs 68%,p = 0.17). Among theselast subjects who underwent polygraphy 50% had OSAS. The preva-lence of an OSAS was significantly higher in patients with, than in

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hose without, F2-F4 fibrosis compared to those without(72% vs4%, respectively; p = 0.04). After correction for confounders, sig-ificant fibrosis was associated with S02 mean levels≤95% meanxyhemoglobin saturation(SaO2) levels < 95%(OR 3.21,95%C.I. 1.02-.34;p = 0.04). Similarly, the prevalence of OSAS was slightly higher

n patients with, than in those without, carotid plaques comparedo(64% vs 40%;p = 0.08). After correction for confounders, the asso-iation between carotid plaques were associated with T90(timepent with SaO2 < 90%) >1% was maintained(OR 6.30,95%C.I.1.02-2.3;p = 0.01).

Conclusions: In NAFLD patients without morbid obesity OSASas highly prevalent and indexes of oxygen saturation were inde-endent indicators of the severity of liver fibrosis and of carotidtherosclerosis risk. These data, if further validated, could suggesto look at for OSAS in all NAFLD patients, considering OSAS as aotential additional therapeutic target for NAFLD.


-13

FNL4 POLYMORPHISMS PREDICT SUSTAINEDESPONSE AND HBSAG CLEARANCE IN

NTERFERON TREATED HBEAG NEGATIVEHRONIC HEPATITIS B PATIENTS

. Galmozzi 1, P. Lampertico 1, F. Facchetti 1,

. Invernizzi 1, G. Mangia 1, M. Vigano 2,. Soffredini 1, M. Colombo 1

Division of Gastroenterology and Hepatology,ondazione IRCCS Cà Granda Ospedale Maggioreoliclinico, University of Milan, Milan, ItalyLiver Unit, Ospedale San Giuseppe, University ofilan, Milan, Italy

Introduction and Aim The recently identified interferonambda 4 (IFNL4) gene harbors the dinucleotide variants368234815-TT/�G, a genetic marker of outcome to IFN-basedherapy of HCV infection. The IFN�-4 protein, which can be gen-rated only by carriers of the rs368234815-�G allele, is thoughto counteract IFN responsiveness by inducing a weak expressionf interferon-stimulated genes. Three nonsynonymous variantsf the IFNL4 gene (rs73555604, rs142981501 and rs117648444)ould affect the IFN�-4 protein. We aimed to explore whetherFNL4 polymorphisms impact on response to IFN-based treatmentn the setting of chronic hepatitis B (CHB).

Materials and Methods IFNL4 gene was sequenced by Sangerethod on genomic DNA extracted from whole blood of 126BeAg-negative CHB patients treated with either standard oregylated-IFN-� and followed-up for 11 years (range 1-17) post-reatment. Results Sustained response rates to IFN were notignificantly different between the 62 carriers of the rs368234815-T/TT (IFN�-4 eliminating) genotype and the 64 carriers of thes368234815-�G (IFN�-4 generating) allele (31% vs 17%, p = 0.079).ince the only nonsynonymous variant identified in our cohort, thero70Ser rs117648444-C/T polymorphism, was exclusively associ-ted with carriers of the IFN�-4 generating allele, these 64 patientsere stratified into rs117648444-CC (IFNl-4 wild-type, n = 45) and

s117648444-CT/TT (IFNl-4 mutated, n = 19) genotypes. Sustainedesponses among IFN-�4 eliminating (rs368234815-TT/TT) andFN-�4 mutated (rs117648444-CT/TT) genotypes (n = 81) were sig-ificantly higher than those in the 45 IFN�-4 wild-type subjects33.3% vs 9%, OR = 4.8, 95%CI 1.6-15.0, p = 0.006). Yet, in the multi-
ariate analysis the combination of IFN�-4 eliminating and IFN�-4utated genotypes independently predict both sustained response
o interferon (OR = 5.33, 95%CI 1.7-16.8, p = 0.004) and off treatment

sease 47S (2015) e43–e66 e49

HBsAg clearance (HR = 4.3, 95%CI 1.5-12.3, p = 0.007). Pretreatmentserum HBV-DNA was the only other independent predictor ofHBsAg loss (HR = 0.61, 95%CI 0.43-0.87, p = 0.007).

Conclusions IFNL4 polymorphisms independently predict sus-tained response to interferon and off treatment HBsAg clearance inHBeAg negative CHB patients.


F-14

NON-INFECTIOUS CO-MORBIDITIES IN HIVPATIENTS CO-INFECTED WITH HEPATITISVIRUSES: AN ANALYSIS FROM THE CALABRHIVSTUDY GROUP

M.C. Postorino 1, F. Luciani 2, C. Mangano 3,M.S. Carpentieri 3, P. Scerbo 4, A. Priamo 4,G. Berardelli 5, R. Marino 6, A. Vallone 6,N. Serrao 7, V. Pisani 1, C. Costa 1, A. Terremoto 2,G. Foti 3, L. Cosco 4, M. Calderazzo 5,D. Corigliano 5, P. Scordo 1,C. Torti 1, and the CalabrHIV Study Group

1 Unità Operativa di Malattie Infettive, AziendaOspedaliera Universitaria “Mater Domini”,Università “Magna Graecia” Catanzaro, Italy2 Unità Operativa di Malattie Infettive, AziendaOspedaliera Cosenza, Italy3 Unità Operativa di Malattie Infettive, AziendaOspedaliera “Bianchi Melacrino Morelli” ReggioCalabria, Italy4 Unità Operativa di Malattie Infettive, Ospedale“Pugliese” Catanzaro, Italy5 Unità Operativa di Malattie Infettive, PresidioOspedaliero Lamezia Terme, Italy6 Unità Operativa di Malattie Infettive, Ospedale“Jazzolino” Vibo Valentia, Italy7 Unità Operativa di Malattie Infettive, OspedaleCrotone, Italy

Introduction and Aims HIV infected patients suffer from pre-mature aging, putting them at risk of non-infectious co-morbiditiesat younger ages than the general population. Aim of this studyis to evaluate for the first time prevalence of non-infectiousco-morbidities and of multi-morbidity in HIV/HCV co-infectedpatients with respect to HIV mono-infected ones.

Materials and Methods The CalabrHIV observational cohortincludes all HIV patients followed by infectious disease centersin the Calabria Region. Epidemiological, clinical and demo-graphic characteristics were collected in a common database.Non-infectious co-morbidities were recorded: cardiovascular dis-eases, hypertension, diabetes, renal failure and bone fractures.Multi-morbidity was defined as ≥2 non infectious co-morbiditiesoccurring in the same patient.

Results 549 patients were selected (69% males, 37% >50 years).Main risk factors were sexual promiscuities (50%) and IVDU (34%).34% patients were HIV/HCV co-infected, 7% HBsAg carriers and2% had triple co-infections. Hypertension was most frequent (25%overall). Multi-morbidity was higher from 50 years of age (30%vs. 6%; p < 0.0001). HIV co-infected patients had more frequentlyAIDS (39% vs. 21%; p < 0.0001). A statistically significant differ-ence in multi-morbidity percentage was observed in patients aged>50 years with HCV and/or HBV co-infection than in HIV mono-
infected ones (70% vs. 50%; p = 0.0037) (Figure 1). This effect wasnot driven by the oldest subjects since an increasing significancewas found with increase in age starting from 40 years (39.5% vs.

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6%; p = 0.0293) and, at >60 years of age, the difference was evennverse (54% vs. 92%; p = 0.026).

Conclusions Even though co-infected patients are no longerspecial” with regard to HCV-treatment response, they are stillfragile”. Prevention and early detection of non infectious co-orbidities is important in these patients, especially in the young.


-15

HIGH HBSAG GENETIC COMPLEXITY CANNFLUENCE HBV IMMUNOGENICITY IN THEETTING OF ACUTE INFECTION

. Battisti 1, M. Aragri 1, N. Coppola 2, C. Alteri 1,

. Sagnelli 3, M. Pisaturo 2, M.C. Bellocchi 1,

. Salpini 1, M. Starace 2, D. Armenia 1, L. Carioti 1,. Pollicita 1, E. Sagnelli 2, C.F. Perno 1, V. Svicher 1

Department of Experimental Medicine and Surgery,niversity of Rome “Tor Vergata”, Rome, ItalyDepartment of Mental Health and Public Medicine,ection of Infectious Diseases, Second University ofaples, Naples, ItalyDepartment of Clinical and Experimental Medicinend Surgery, Second University of Naples, Naples,taly

Background. To characterize HBV RT and HBsAg genetic-eterogeneity in acute HBV (AHB) infected patients and to defineheir clinical value.

Methods. 62 HBsAg + and IgM/anti-HBc+ patients with clinicalnd biochemical signs of AHB (44 genotype-D and 18 genotype-A)ere enrolled from 2000 to 2010. Plasma-samples obtained at first-

bservation were analyzed by ultra-deep sequencing (UDPS) forrug-resistance and immune-escape mutations. Shannon-Entropy,eighted for intra-patient prevalence of each mutated-position,as used to measure the extent of HBsAg amino-acid variability.

Results. 75.8% of patients were male with median(IQR)ge of 36(29-46) years. Median(IQR) ALT and HBV-DNA were,544(1,938-3,078)U/L and 5.9(4.5-7.4)log10IU/ml. 61/62 (98.4%)
atients became HBsAg-negative with 33/61 (54.1%) developinglso anti-HBs.
By UDPS, 8.1% (5/62) of patients carried ≥1 drug-resistanceutation (rtV173L/rtL180 M/rtA181 T/rtA194 T/rtM204I). They

sease 47S (2015) e43–e66

were detected with an intra-patient prevalence ranging from0.11% for rtA181 T to 99.98% rtL180 M.

Analysing HBsAg a-determinant, 48.4% (30/62) of patientscarried ≥1 immune-escape mutation (intra-patient prevalencerange:0.2-100%). Vaccine-escape mutations were found in 11.4%of patients, all genotype D-infected. This is the case of sG145R,sM133L, and sP120S, detected with intra-patient prevalence ran-ging from 3.9% to 99.9% for sG145R, from 1.9% to 16.8% for sM133L,and always of 100% for sP120S.

Stop-codons were found in 19.3% patients (intra-patient preva-lence range:1.6-47.5%). They occurred at 11 HBsAg-positionsincluding also 172 and 182, known to increase HBV oncogenic-potential.

Finally, by Shannon-Entropy, specific HBsAg-positions corre-lated with the lack of HBsAg seroconversion in genotype-D. Inparticular, positions 130 and 133 (localized in HLA-class-II epi-tope) were found mutated only in patients not developing anti-HBs(Shannon-Entropy mean ± SE:1.98 ± 0.01 vs 0, and 1.95 ± 0.03 vs 0,respectively, P < 0.05).

Conclusions: AHB is characterized by a complex coexistenceof viral-quasispecies, some of them with reduced antigenic-ity/immunogenicity, enhanced oncogenic-potential and altereddrug-susceptibility. These viral-variants may induce severe and/ordifficult-to-treat forms of HBV-infection (es. HBV-reactivation),and might affect the efficacy of current HBV-vaccination strategy.


F-16

UPDATED CUT OFF VALUES OF LIVER STIFFNESSTO MAXIMIZE TREATMENT OUTCOME OFINTERFERON FREE ANTI HCV REGIMENS

A. Colli 1, M. Fraquelli 2, D. Prati 3, A. Aghemo 4,D. Conte 2, M. Colombo 4, G. Casazza 5

1 Dipartimento di Medicina Interna, Ospedale AManzoni, Lecco, Italy2 Unità di Gastroenterologia ed Endoscopia,Fondazione IRCCS Ospedale Maggiore Policlinico,Università degli Studi di Milano, Milano, Italy3 Dipartimento di Medicina Trasfusionale edEmatologia, Ospedale A Manzoni, Lecco, Italy4 Unità di Gastroenterologia ed Epatologia,Fondazione IRCCS Ospedale Maggiore Policlinico,Università degli Studi di Milano, Milano, Italy5 Dipartimento di Scienze Biomediche e Cliniche,Ospedale Luigi Sacco, Milano, Italy

BACKGROUND Interferon-free anti-HCV regimens will changethe paradigm of hepatitis C treatment.Yet due to money constrainstreatment access relies on pretreatment patients stratification fordisease severity using non-invasive tests including liver stiffnessmeasurement (LSM), whose cut-off values have been establishedto maximize its diagnostic accuracy.Now cut offs are required tomaximize the clinical benefit of treatment. Based on the preva-lence of different fibrosis stages and the harm/benefit ratio(H/B)of different treatments, cut-off values can be obtained to drive thetreatment decision, i.e.to identify patients with the minimum levelof hepatitis requiring treatment.

METHODS 11 hepatologists in different tertiary centersquantified the expected H/B, based on published data and clin-ical judgment, for different scenarios: two different disease
stages (F≥2orF≥3) for threetreatment regimens (PEGIFN + RBVor PEGIFN + RBV + 1STgeneration PI or IFNfree regimens). From acohort of 750 consecutive HCV patients we obtained two ROC

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urves assessing LSM accuracy in diagnosing F≥2 or F≥3 (RS:liveriopsy). The prevalence,p(D), of F ≥2 or F ≥3 (D) was respectively3% and 30%. For each scenarios the optimal cut-off value maxi-izing the expected utility of LSM was identified on the ROC curve

s the point with slope equal to equation(1). Normal median LSMalues in a control population (#1001 #participants) from the samerea was 4.7 kPa (2.6-7.6, percentile 5th - 95th).

RESULTS Mean H/B value for the three different therapeuticegimens according to two different liver disease scenarios:

PEGINF +RBVF≥2

F≥3 PEGINF +RBV + PIF≥2

F≥3 IFN-freeF≥2

F≥3

Mean H/B 1/3.1 1/3.7 1/2.8 1/4.1 1/8.3 1/10LSM(kPa)cut-offs

6.1 9.4 6.4 9.3 4.1 7.1

CONCLUSIONS The estimated H/B of interferon-free regimens isxtremely low and thus the LSM cut-off values to initiate treatmentre lower than the ones predicting maximal diagnostic accuracy,ither assuming to treat patients with F≥3 or all comers with F≥2.

1)Cantor et al. J Clin Epidemiol 1999


-17

AN HEPATIC RESECTION PROVIDE AONG-TERM CURE TO PATIENTS WITHNTRAHEPATIC CHOLANGIOCARCINOMA?

. Spolverato 1,∗, A. Vitale 2,∗, A. Cucchetti 3,. Alexandrescu 4, H.P. Marques 5, L. Aldrighetti 6,.C. Gamblin 7, S.K. Maithel 8, C. Pulitano 9,.W. Bauer 10, F. Shen 11, G.A. Poultsides 12,.W. Marsh 13, T.M. Pawlik 1

The Johns Hopkins University School of Medicine,altimore, MD, United States of AmericaUnità di Chirurgia Epatobiliare e Trapianto Epatico,zienda Ospedaliera-Università di Padova, Padova,

talyDepartment of Medical and Surgical Sciences, S.rsola-Malpighi Hospital, Alma Mater Studiorum -niversity of Bologna, Bologna, ItalyFundeni Clinical Institute, Bucharest, RomaniaCurry Cabral Hospital, Lisbon, PortugalOspedale San Raffaele, Milan, ItalyMedical College of Wisconsin, Milwaukee, WI,nited States of AmericaEmory University, Atlanta, GA, United States ofmericaUniversity of Sydney, Sydney, Australia

0 University of Virginia, Charlottesville, VA, Unitedtates of America1 Eastern Hepatobiliary Surgery Hospital, Shanghai,hina2 Stanford University, Stanford, CA, United States ofmerica3 University of Pittsburgh Medical Center,ittsburgh, PA, United States of America

Background: A patient can be considered statistically curedrom the specific disease when the mortality returns to the sameevel as that of the general population. Cure models have never been
pplied to patients undergoing hepatic resection for intrahepaticholangiocarcinoma (ICC). We sought to assess the probability ofeing statistically cured from ICC by hepatic resection.
sease 47S (2015) e43–e66 e51

Methods: 584 patients undergoing surgery with curative intentfor ICC between 1990 and 2013 at one of 12 participating insti-tutions were identified. A nonmixture cure model was adopted tocompare mortality after hepatic resection to the mortality expectedfor the general population matched by sex and age.

Results: Median, 1-, 3-, 5- years disease-free survival was 10months, 44%, 18% and 11%; the corresponding overall survival was27 months, 75%, 37% and 22%. The probability of being cured fromICC was 9.7% (95% confidence interval 6.1-13.4%). The mortality ofpatients undergoing surgery for ICC was higher than the generalpopulation until year 10, when the patients alive without tumorrecurrence can be considered cured with 99% certainty. Multivari-ate analysis showed that cure probabilities range from 25.8% (timeto cure 9.8 years) in patients with single ICC ≤ 5 cm, and withoutminor vascular invasion, poor grade, lymph nodes metastases, andperiductal histology, to < 0.1% (time to cure 12.6 years) in patientswith all these six risk factors.

Conclusions. The cure model indicates that statistical cure ispossible in patients undergoing hepatic resection for ICC. A modelto calculate the cure fraction and time to cure in each clinical sce-nario was provided (http://www.livercancer.eu/CCC cure model).


F-18

AUTOFLUORESCENCE DETECTION OF LIVEROXIDATIVE DAMAGE PRODUCTS

A.C. Croce 1,2, A. Ferrigno 3, V.M. Piccolini 1,L.G. Di Pasqua 3, C. Berado 3, G. Bottiroli 1,2,M. Vairetti 3

1 IGM-CNR, Pavia, Italy2 Department of Biology & Biotechnology, UniversityPavia, Italy3 Department of Internal Medicine and Therapy,University of Pavia, Pavia, Italy

Introduction- Oxidative stress in fatty livers enhances the riskof disease progression or organ dysfunction in surgery and trans-plantation. An experimental model of induced-NASH showed aliver metabolism subversion, reflected by tissue autofluorescence(AF) alterations. These consisted in changes in NAD(P)H/flavin AFrelated contributions -ascribable to redox metabolism alterations-and a rising in vitamin A, protein, and lipofuscin-like-lipopigmentemission signals.

Aim- Lipofuscin-like-lipopigments derive from the oxidation ofunsaturated lipid. Therefore fluorescing fatty acids and oxidizedproducts were investigated as early oxidative biomarkers in liverswith mild lipid accumulation, under different preservation condi-tions.

Materials and Methods- Rat livers(controls and 2 weekadministered methionine/choline-deficient diet-MCD) were iso-lated and submitted to 6-h Cold Storage (CS) or subnormotermicMachine Perfusion (MP), followed by reperfusion (37 ◦C, oxy-genated medium). AF spectra recorded in vivo (exc 366 nm), viafiber-optic probe, were analyzed by curve fitting procedure, toestimate each endogenous fluorophore contribution to the overallemission, similarly to in situ biochemical analysis. Tissue oxida-tive stress (TBARS, and GSH) and mitochondrial dysfunction(ATP/ADP) were assayed using conventional methods. Results. BothCS or MP preservation affected liver AF properties. The responseof NAD(P)H/flavin redox state to oxygenation and temperature
was consistent with MCD induced mitochondrial dysfunction.Lipofuscin-like-lipopigment AF signals rose in both controls andMCD livers, in a good correlation with TBARS. Independent from

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reservation procedure, the phenomenon was significantly greatern MCD than in control livers, indicating a strong influence of fattyiver oxidative stress. Fluorescing fatty acids were not depleted,onsistently with dynamic equilibria in liver lipid pool composition.

Conclusions. The real time AF analysis allows to monitor oxida-ive effects even in a model of mild metabolic alterations, providingn AF support in experimental investigations on drug response andoxicity, and in the set-up of innovative organ preservation strate-ies.(Supported by Fondazione Cariplo, grant n◦2011-0439).


-19

REVALENCE AND RISK FACTORS OF METABOLICYNDROME AFTER LIVER TRANSPLANTATION: AINGLE CENTRE EXPERIENCE

. Pepe 1, G. Germani 1, A. Ferrarese 1, A. Zanetto 1,. Bortoluzzi 1, E. Nadal 1, F.P. Russo 1,

. Senzolo 1, E. Gringeri 2, U. Cillo 2, P. Burra 1

Multivisceral Transplant Unit, Department ofurgery, Oncology and Gastroenterology, Paduaniversity Hospital, Padua, ItalyHepatobiliary Surgery and Liver Transplant Unit,adua University Hospital, Padua, Italy

Background and Aim: Metabolic syndrome (MS) is commonfter liver transplant (LT), increasing long term mortality and mor-idity. The aim of this study was to evaluate short and long-termrevalence and risk factors of MS after LT.

Materials and methods: patients who underwent LT at Padovaiver Transplant Center between January 2000 and March 2013retrospective cohort) and between April 2013 and April 2014prospective cohort) and followed-up at Multivisceral Transplantnit were included. Patients < 18 years, who underwent multior-an transplantation or re-LT and with pre-LT MS were excluded.or each patient general and metabolic variables and donor char-cteristics were recorded.

Results: In the retrospective cohort 161 patients (120 M) werencluded. The most common indication to LT was HCV-relatedirrhosis (49.1%). A post-LT significant increase in BMI values, iniabetes and hypertension prevalence and in total cholesterol andriglyceride levels was found compared to pre-LT values. At a meanost-LT follow-up of 6.9 ± 4.2 years 81/161 (50.3%) patients devel-ped MS. Recipient male sex (OR 2.36, p = 0.045), a higher pre-LTMI (OR per unit 1.14, p = 0.03), and the presence of pre-LT diabetesOR 5.98, p = 0.04) were associated with the development of post-T MS. In the prospective cohort 15 patients were included (10 M).t 3, 6 and 12 months after LT a significant increase in BMI val-es, diabetes and hypertension prevalence and in cholesterol andriglyceride levels was found compared to pre-transplant valuesnd 5/15 (33.3%), 3/11 (27.3%) e 4/10 (40%) patients developed MS.

Conclusions: post-LT MS affects nearly half of LT recipients,tarting early after LT. Recipient male gender, pre-LT diabetes andncreased BMI are risk factors for MS after LT. Lifestyle modifica-ions, should be recommended to transplanted recipients startingn the early post-LT period, thus preventing body weight gain and
he associated abnormalities and reducing incidence of post-LT MSnd the related cardio-vascular events.

sease 47S (2015) e43–e66

F-20

SHORT-TERM OUTCOME POST LIVERTRANSPLANTATION OF CIRRHOTIC LISTEDOUTPATIENTS WITH FLUCTUATIONS OF EGFR

F. Fiacco 1, F. Tinti 1, I. Umbro 1, A. Zavatto 1,S. Ginanni Corradini 2, M. Rossi 3, P.B. Berloco 3,A.P. Mitterhofer 1

1 Department of Clinical Medicine, Nephrology andDialysis Unit, Sapienza University of Rome, Rome,Italy2 Department of Clinical Medicine, GastroenterologyUnit, Sapienza University of Rome, Rome, Italy3 Department of General Surgery, Organ TransplantUnit “Paride Stefanini”, Sapienza University of Rome,Rome, Italy

Introduction-In cirrhotic patients listed for liver transplanta-tion (ESLD-wLT) renal dysfunction results from systemic conditionsthat affect both liver and kidney leading to kidney ischemia. Thesepatients are prone to estimated-glomerular filtration rate (eGFR)changes related to pre-renal conditions. Nephrologic guidelinesconsider eGFR modifications common and not necessarily indica-tive of progression to chronic kidney disease (CKD), but theirmeaning in ESLD-wLT outpatients is not clear and assessment ofeGFR is challenging.

Aim-to evaluate eGFR variations in ESLD-wLT outpatients andoutcome post-LT related to short-term renal, graft and patient sur-vival.

Methods-Single centre retrospective study of 51 ESLD-wLT out-patients. All available GFR values estimated by MDRD formula werecollected considering eGFR at listing (eGFR-list), eGFR at transplant(eGFR-LT), the lowest (eGFR-min) and the highest value of eGFR,further defined as baseline (eGFR-baseline). Fluctuations (FeGFR)were considered as percentage variation between eGFR-baselineand eGFR-min. Fluctuations of eGFR > 50% were defined as drop.Acute Kidney Injury (AKI) and CKD were defined according toKDIGO-Guidelines. Short-term graft function, AKI post-LT and 30-days survival after-LT were assessed.

Results-All patients presented FeGFR. Estimated-GFR-LT wasnot different from eGFR-list. Estimated-GFR drops were observedin 18/51 patients (DeGFR + group). These patients presented higherprevalence of pre-LT CKD, higher MELD score and bilirubin both atlisting and LT and lower sCr-min. AKI post-LT stages 2-3 occurredmore frequently in DeGFR+ group compared to DeGFR- group (8/18vs 3/33,p0.01). Patients in DeGFR+ group required more plasmainfusions and inotropic support during LT, while no differenceswere detected in early graft function and survival after-LT.

Conclusions-Estimated-GFR fluctuations are common in ESLD-wLT outpatients and are not indicative of worsening of renalfunction from listing to LT. Estimated-GFR drops are more frequentin patients with CKD and worse condition of cirrhosis and are asso-

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-21

IRST STEPS TOWARDS UNDERSTANDING THEYNAMIC EVOLUTION OF GUT MICROBIOTA INIFFERENT STAGES OF LIVER DISEASE

.R. Ponziani 1, S. Pecere 1, F. Paroni Sterbini 2,. Petito 1, M. Siciliano 1, T. Di Rienzo 1,. Palladini 2, D. Zambrano 1, F. Franceschi 1,. Gaetani 1, F. Scaldaferri 1, L. Masucci 2,. Sanguinetti 2, A. Gasbarrini 1

Internal Medicine Department, Gastroenterologyivision, Catholic University Of Sacred Heart ofome, Rome, ItalyMicrobiology Department, Catholic University Ofacred Heart of Rome, Rome, Italy

INTRODUCTION: Liver cirrhosis is associated to intestinal bar-ier derangement and dysbiosis. AIM: To preliminary describe guticrobiota modifications in a population of Italian patients with

iver cirrhosis.PATIENTS AND METHODS: Fecal samples were collected in

3 cirrhotic patients and 10 healthy controls, no exposed fort least one month to antibiotics, prebiotics, probiotics andowel colonoscopy preparation. Gut microbiota composition wasssessed by a metagenomic gene-targeted approach (16S rRNA),ollowing DNA isolation from stool samples stored at–80 ◦C. Dataere analyzed in Qiime. Non parametric tests were used for thenal statistical analysis.

RESULTS: The enterotypification of the 23 subjects was compa-able to that of the Western general population, with a prevalence ofacteroides (enterotype 1, 77%, often overlapping with enterotype, 18%) with no inter- or intra-group differences between cirrhoticsnd healthy controls. A trend toward an unbalance between the Fir-icutes to Bacteroidetes ratio was found throughout the different

hild Pugh stages (close to 1 in Child A, reduced to half in Child Bnd to ¼ in Child C p = 0.081). Compared to healthy individuals, cir-hotics microbiota was more enriched in Bacilli (Lactobacillaceae,treptococcaceae, Enterococcaceae; p = 0.001), especially in case ofntermediate stage liver disease (abundance in Child B patients8.6%; p = 0.008). Clostridia (Clostridiaceae, Ruminococcaceae,eillonellaceae, Lachnospiraceae, Clostridiaceae, Peptostreptococ-aceae, Peptococcaceae) were the most abundant in the initialhases of the disease (62.3% in Child A patients p = 0.06). Patientsith at least one clinical sign of decompensation lost the rela-

ive abundance of Clostridia (abundance 62.3% in compensatedatients vs 20.8% in decompensated ones vs 35.4% in healthy con-rols p = 0.04).

CONCLUSION: Liver cirrhosis is a fascinating model of gut micro-
iota modifications, that are connected with the progression of liver
mpairment.


sease 47S (2015) e43–e66 e53

F-22

PREVALENCE OF HEPATITIS E IN TERTIARYHOSPITAL CENTRE IN NORTHERN ITALY

V. Zuccaro 1, P. Columpsi 1, S. Paolucci 2,M. Mariani 1, S. Toppino 1, A. Malfitano 1,A. Parisi 1, S.F.A. Patruno 1, F. Baldanti 2, R. Bruno 1

1 Department of Infectious Diseases–IRCCS SanMatteo - University of Pavia, Pavia, Italy2 Virology Unit IRCCS San Matteo - University ofPavia, Pavia, Italy

Introduction: In developed countries, hepatitis E is a zoonosisusually caused by the genotypes 3 and 4. In Italy the prevalence ofanti-HEV is 1-5% among the healthy population. A large prospectivestudy has shown that HEV accounts for approximately 10% of acutenon A, non B hepatitis cases in Italy.

Aim: The aim of the study was to evaluate the prevalence of hep-atitis E among patients admitted to the Division of Infectious andtropical Disease for acute hepatitis without evidence of autoimmu-nity and antibody against HAV-HBV-HCV.

Materials and Methods Results: A total of 970 patients wereadmitted to Division of Tropical and Infectious Diseases from 2013to 2014. Among them 27 had acute hepatitis and 3 (11%) were pos-itive for hepatitis E. All were anti-HEV IgM and IgG positive andwere also positive for HEV-RNA, viral genotype was 3 subtype Fin all cases. All HEV patients were living from Pavia and deniedhaving travelled abroad. Diagnosis of hepatitis E was based on thepresence of IgM anti-HEV and confirmed by the detection of HEV-RNA - RTPCR on patients samples. Viral isolates were sequencedto characterize subtypes. Two cases with acute disease had a self-limited course with AST-ALT normalization within 3-6 weeks. Onepatient died from acute on chronic liver disease. All cases showedhigh level of amylases during the acute phase.

Conclusions: This is the first report of hepatitis E in thePavia area and it confirms the increasing trend of prevalence ofautochthonous hepatitis E in our country. The increased prevalencemight be related either to the spread of the HEV virus in our coun-try and to the implementation of HEV screening in patients withacute hepatitis. In the case of acute hepatitis with high amylaseslevel should prompt exclude HEV infection.


F-23

IMPACT OF RIFAXIMIN IN THE PREVENTION OFBACTERIAL INFECTIONS IN CIRRHOSIS

M. Mariani 1, L. Scudeller 2, S.F.A. Patruno 1,V. Zuccaro 1, P. Columpsi 1, S. Toppino 1,A. Malfitano 1, A. Parisi 1, G. Filice 1, R. Bruno 1

1 Division of Infectious and Tropical Diseases–IRCCSSan Matteo - University of Pavia, Pavia, Italy2 Scientific Direction, IRCCS Policlinico San Matteo,Pavia, Italy

Background and aims: Infectious events represent a leadingfactor able to accelerate the progression from compensated todecompensated stage of cirrhosis, with a subsequent worseningof the prognosis. Due to the high frequency of infectious compli-cations in cirrhotic patients, a great effort was made in order to
improve the survival rate. Rifaximin was advanced as a possibleprimary prophylactic therapy against spontaneous bacterial infec-tions. We retrospectively investigated the role of this antibiotic in

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educing the proportion of infected patients in treated and non-reated groups.

Methods: We enrolled 649 patients whose clinical and personalata, prescribed therapy, microbiological findings and laboratoryests were collected from previous discharge letters and our institu-ion database. We excluded patients with ACLF and acute hepatitis.etiology, drug resistance and site of infection were collected asell.

Results: Risk of developing bacterial infections was significantlyower in patients treated with rifaximin (6 month of treatment, 10ays per month, 400gr t.i.d.) (OR 0.29; 95% CI 0.20-0.40, p < 0.001)ompared to those not treated. Moreover, in our cohort we findut a prevalence of infections sustained by Gram-positive bacteria61.5% of isolations).

Conclusions: Rifaximin continuous treatment turned out to bevery effective factor involved in bacterial infections prevention

n cirrhotic patients. Our study showed a shift in general bacterialrevalence among pathogens to Gram positive, this finding shoulde considered as well before administering an empirical therapy inirrhotic patients.


-24

ERUM ALPHA-FETOPROTEIN MODIFICATIONDELTA-AFP) IS A HIGHLY SPECIFIC TOOL TOUSPECT HCC RECURRENCE AFTER LIVERRANSPLANTATION

.R. Ponziani 1,∗, S. Bhoori 1, M. Bongini 1,. Flores 1, C. Muscarà 1, V. Mazzaferro 1

Liver Transplant, Hepatobiliary andastrointestinal Surgery, Hepatology Nationalancer Insitute of Milan, Milan, Italy

Introduction: Alpha-fetoprotein (AFP) is the main tumorarker of hepatocellular carcinoma (HCC). Despite AFP values at

isting have been used to predict HCC recurrence after liver trans-lantation (LT), its role in patients’ surveillance after LT has noteen established yet.

Aim: To investigate the usefulness of serum AFP modificationo suspect recurrence in patients who previously underwent LT forCC.

Materials and Methods Results: We investigated all consecu-ive patients undergoing LT for HCC within Milan or up-to-sevenriteria from January 2004 to December 2013. Serum AFP wasollected at the time of LT and every 6-months for 2 years andvery year thereafter. Patients surveillance consisted in semian-ual abdominal ultrasound or CT scan for the first 5-years, thenvery year.

231 LT recipients were investigated (median age 61 (14-64), 89%ales, 30% HBV, 51.5% HCV, 4% coinfected, 10% alcohol, 8% other,

% HIV); 31 of them (13%) experienced HCC recurrence. AmongCC recurrence-free patients, 24 (10%) developed cirrhosis or liver

ailure, severe hepatitis or alcohol abuse during follow-up; sinceheir median AFP values had higher peaks, they were excluded tovoid biases. Univariate analysis failed to demonstrate any asso-iation between serum AFP at any timepoint and HCC recurrence,hile its modification (difference between the highest AFP valueetected during follow-up or the value detected at recurrence,nd the lowest serum AFP value, named “delta-AFP”) was stronglyssociated with HCC recurrence (p < 0.0001). Multivariable analy-
is confirmed delta-AFP as the only independent predictive factorf HCC recurrence (OR 1.28, 95%CI 1.116, 1.470; p = 0.001). Theccuracy of delta-AFP in predicting recurrence was 71.5% (95%CI
sease 47S (2015) e43–e66

65% > 77.5%), with the best identified cut-off value of 6.57 ng/ml(sensitivity 61.3% specificity 100%, NPV 96% PPV 100%).

Conclusions: Delta-AFP is a very easy-to-use and highly specifictool to suspect HCC recurrence after LT.


F-25

CLINICAL PRESENTATION, TREATMENT ANDOUTCOME OF ALCOHOL- AND HEPATITIS CVIRUS- RELATED HEPATOCELLULARCARCINOMA IN THE NEW CENTURY:COMPARISON BEFORE AND AFTER ADJUSTMENTWITH PROPENSITY SCORE ANALYSIS

L. Bucci 1, F. Garuti 1, V. Camelli 1, B. Lenzi 1,M. Bernardi 1,F. Trevisani 1, for the Italian Liver Cancer (ITA.LI.CA) group

1 Dipartimento di Scienze Mediche e Chirurgiche -Alma Mate Studiorum - University of Bologna,Bologna, Italy

Introduction and aims: Hepatitis C virus (HCV) and alcoholabuse are the main causes of hepatocellular carcinoma (HCC) inWestern world, accounting for about 60% and 20% of cases, respec-tively. This study aimed at comparing the clinical presentation andoutcome of alcohol- and HCV- related HCC diagnosed in the newcentury.

Materials and Methods: 1844 HCV and 636 alcoholic patientsfrom the Italian Liver Cancer (ITA.LI.CA) database, diagnosed withHCC from January 2000 to December 2012 were compared for age,gender, type of diagnosis, Barcelona Clinic Liver Cancer (BCLC),tumor gross pathology, portal vein thrombosis (PVT), esophagealvarices, model for end stage liver disease (MELD), Child-Pugh class,alpha-fetoprotein (AFP), treatment and survival. For survival anal-ysis, a propensity matching model was performed to minimizeimbalances of prognostic factors between groups.

Results: Underlying cirrhosis was present in 96% of bothalcoholic and HCV patients. Alcoholic patients were significantlyyounger and more likely male, diagnosed with a tumor outsidesurveillance, in intermediate and terminal BCLC stage and in aworse hepatic function setting. Consequently, the overall survivalwas decreased in alcoholic compared to HCV patients [median(95% C.I.): 34.5 (28.7-40.2) vs 43.6 (40.9-46.3) months; p < 0.001]. Aone-to-one matching after propensity score adjustment accordingto age, gender, type of diagnosis, BCLC stage, tumor gross pathol-ogy, PVT and treatment was performed. This analysis selected 354patients per group, well balanced for baseline prognostic factors.The overall survival did not differ any longer between alcoholic andHCV group [median (95% C.I.): 42.6 (36.5-48.8) vs 38.6 (31.2-45.9)months; p = 0.756].

Conclusions: Alcoholic patients present with a more compro-mised liver function and a more advanced HCC, less frequentlydetected during surveillance. Nevertheless, alcoholic etiology perse does not affect survival as compared with HCV-related cases,

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-26

NOS DYSFUNCTION IN STEATOSIS ANDTEATOHEPATITIS

. Masarone 1, A. Carrizzo 2, A. Federico 3,. Rosato 4, E. Claar 5, C. Vecchione 6, M. Persico 1

Internal Medicine and Hepatology Unit, Universityf Salerno, Salerno, ItalyVascular Physiopathology Unit IRCCS, INMeuromed, Pozzilli (IS), ItalyGastroenterology Division - Second University ofaples, ItalyInternal Medicine and Hepatology Department,econd University of Naples, ItalyInternal Medicine Department, Hepatology Unit,spedale Evangelico Villa Betania, Naples, ItalyDepartment of Medicine and Surgery, University ofalerno, Salerno, Italy

Background: Non-Alcoholic-Fatty-Liver (NAFLD/NASH) haseen associated to cardiometabolic syndrome that is a riskactor for cardiovascular diseases (CVD). The basis of CVD isxidative stress. Endothelial Nitric-Oxide-Synthase (eNOS) dys-unction has been widely involved in oxidative stress and CVD.lthough NAFLD/NASH and CVD are associated to the same fac-

ors, NAFLD/NASH represents itself an independent CVD risk-factor.ven diabetics show higher rates of CVD if they have NASH. eNOSerangement has been demonstrated in animal experimental mod-ls of NAFLD/NASH. Although clinical and “sublinical” markersi.e.“intima-media-thickness”) seem to have confirmed this sus-icion, nevertheless, no experimental studies on humans haveirectly demonstrated that eNOS dysfunction is associated withAFLD/NASH.

Aim: to directly demonstrate eNOS derangement inAFLD/NASH.

Patients and methods: 18 patients (13 M,5F), coming in ourepartment of Internal Medicine for NAFLD/NASH, were consecu-ively enrolled. Every patient underwent a clinical evaluation andliver biopsy after informed consent. Patients were divided in tworoups according to the presence of NAFLD or NASH. We measuredNOS function by evaluating the vasorelaxation activity induced onsolated-mice-vessels by platelet-rich-plasma of peripheral blood,nd by immunoblot-assays for platelet-derived eNOS (p-eNOS).ollected data were compared to those from an age-, sex-matchedroup of healthy volunteers from a local blood bank. All subjectsere non-smokers and had no active CVD.

Results:Of the 18 pts, 7 (38,8%) had NAFLD and 11 (61,7%)ad NASH at the liver biopsy. No statistically significant differ-nces were found between the two groups and controls for age,ex, BMI, ALT, hypertension, diabetes, dyslipidaemia, obesity andetabolic syndrome. Vascular reactivity curves demonstrated a

educed activity of eNOS in patients with NAFLD/NASH in respecto controls(p < 0.005). Moreover, immunoblot-assays for p-eNOSemonstrated a significantly lower expression in NAFLD/NASHatients in respect to controls (p < 0.007).

Conclusions: we directly demonstrated that eNOS function iseduced in NAFLD/NASH. Endothelial dysfunction may be consid-
red as one of the pathophysiological mechanisms of liver damagen NAFLD/NASH.

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F-27

SOFOSBUVIR PLUS RIBAVIRIN IN HCV-INFECTEDCIRRHOTIC PATIENTS AWAITING LIVERTRANSPLANTATION: PRELIMINARY DATA FROMA SINGLE-CENTRE EXPERIENCE

S. Martini 1, S. Strona 1, D. Arese 1, M. Sacco 1,R. Romagnoli 2, F. Tandoi 2, A. Ottobrelli 1,M.R. Torrani-Cerenzia 1, F. Balzola 1, M. Salizzoni 2,M. Rizzetto 1

1 Gastrohepatology Unit, Department of MedicalSciences, A.O.U. Città della Salute e della Scienza diTorino, Turin, Italy2 Liver Transplant Centre, General Surgery Unit,Department of Surgical Sciences, A.O.U. Città dellaSalute e della Scienza di Torino, Turin, Italy

Background and aims: Clearance of HCV infection before livertransplantation (LT) prevents recurrence after LT. We evaluatedsafety and efficacy of antiviral therapy (Tx) with sofosbuvir (SOF)plus ribavirin (RBV) in HCV-infected cirrhotic patients (pts) beforeLT.

Methods: Since June 2014 to date we enrolled 24 HCV cirrhoticpts with HCC within Milan criteria (15 pts) and/or decompensatedliver disease. Median MELD 13 (range 7-19); Child-Pugh score B71% and C 17%; 83% male, mean age 55.4 years, mean BMI 25.8.Mean baseline HCVRNA 6.17 log10 IU/mL (4.91- 7.11 log10 IU/mL);GT1b 55%, GT1a 4%, GT2 4%, GT3 29%, GT4 8%; IL28CC 20%; 63%experienced. Planned treatment: SOF 400 mg/die (compassionateuse) and RBV (weight-based) for 48 weeks or until LT.

Results: Antiviral treatment resulted in rapid suppression of cir-culating virus (median decrease HCVRNA: 3.27 log10 IU/mL after 1week of Tx); 10/20 pts (50%) at week 4, 14/14 week 8, 10/10 week 12and 7/7 week 16 on Tx were HCVRNA negative. No serious adverseevents were observed. RBV reduction and/or epoetin administra-tion was required in 25% of pts and 3 were hospitalized due tocomplications of liver disease. Of the 7 pts transplanted until now,4 had HCVRNA < 15 IU/mL for a median of 62 days (31-91) and dis-continued antiviral Tx at LT. All those 4 pts are currently HCVRNAnegative after a median follow-up post-LT of 45 days (16-69). 2 ptsunderwent LT while still being HCVRNA positive after less than 4weeks of Tx, and the last one was transplanted 3 days ago havingHCVRNA <15 IU/mL since 5 days.

Conclusion. Tx with SOF plus RBV induces rapid HCV clearanceand is well tolerated in cirrhotic pts awaiting LT. Preliminary post-

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-28

ISTRIBUTION OF STEM CELLS AND CANCERTEM CELLS MARKERS IN LIVER PATHOLOGIESND THEIR INDICATION TO THE RESPONSE OFHERAPY

. H.C. Sukowati 1,2, B. Anfuso 1, D. Pascut 1,

. Patti 1,2, N. Mezzina 1,2, P. Tarchi 3,.L. Crocè 1,2, C. Tiribelli 1,2

Fondazione Italiana Fegato, AREA Science Parkasovizza, Trieste, ItalyDepartment of Medicine, Surgery, and Healthciences, University of Trieste, Trieste, ItalyDepartment of General Surgery, Teaching Hospitalattinara, Trieste, Italy

Introduction Recent studies had suggested different markerso identify the population of stem cells (SC) and cancer stem cellsCSC) in hepatocellular carcinoma (HCC). However, a study on theistribution of the markers that may indicate disease progression,ature of cancer, and response to therapy will be crucial in bothasic and translational research.

Aim To study the variation and distribution of various SCnd CSC markers during hepatocarcinogenesis and the associationetween circulating CSC and HCC therapy.

Methods We analyzed 119 samples (12 normal tissues, 40 liverirrhosis (LC), 33 HCC; 34 blood samples of HCC). The comparisonf SC and CSC markers CD90, CD44, CD133, EpCAM, CD13, CK19,K7, and ABCG2 in each pathology groups was assessed by RTq-CR using MIQE guidelines. The effect of different treatments asesection, radiofrequency, and chemoembolization to circulatingSC was assessed by comparing positivity and expression on pre-reatment (T0) with 1 month post-treatment (T1).

Results The expression of CD90 was positively correlated withepatocarcinogenesis moving from normal, LC, to HCC (p < 0.001)onfirmed by Western Blot; an increase in CD44 expression waslso observed. In contrast, the expressions of EpCAM, CD133, CK19,nd CK7 were increased only in LC. The CD13 expression was relatedith HCV and MELD score (p < 0.05) while the ratio HCC:LC ofBCG2 was associated with Edmonson-Steiner grade.

At T0, 80% of blood were positive for CSC CD90 and only 40% foroth CD133 and EpCAM. At T1, HCC resection reduced the expres-ions of CSC markers CD90, CD133, and EpCAM, radiofrequencyeduced CD133, while chemoembolization increased EpCAM.

Conclusion This study demonstrated that CSC CD90 marker isncreased during hepatocarcinogenesis. The detection of circulating
SC may be used to study the response to therapy without takingiopsy, it will be important in the diagnostic and prognostic value.

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F-29

LIVER TRANSPLANTATION (OLT) ANDCARDIOVASCULAR RISK: ASSESSMENT OF EARLYATHEROSCLEROTIC DAMAGE AND METABOLICDISORDERS IN PATIENTS UNDERGOING LIVERTRANSPLANTATION

G. Pisano 1, M. Orlandi 1, F. Donato 2, S. Zannoni 1,C. Bertelli 1, M.Porzio 1, M. Colombo 2, S. Fargion 1,A.L. Fracanzani 1

1 Department of Pathophysiology andTransplantation, Maggiore Policlinico HospitalFoundation IRCCS, University of Milan, Milan, Italy2 Division of Gastroenterology, Maggiore PoliclinicoHospital Foundation IRCCS, University of Milan,Milan, Italy

BACKGROUND: Improved survival after orthotropic liver trans-plantation (OLT) has allowed to detect long-term consequences ofliver transplantation. Nowadays, cardiovascular and cerebrovascu-lar diseases are the leading causes of morbidity and death amongtransplant recipients. Metabolic syndrome and its individual com-ponents, (diabetes mellitus, hypertension, dyslipidemia, obesity)contributing to cardiovascular complications are increasingly beingidentified in these patients.

AIMS: To evaluate the prevalence of metabolic syndrome and ofsubclinical atherosclerosis in patients with OLT at 6 month and at1 year follow-up.

METHODS: We evaluated 42 patients: 27 on transplant list(group 1) and 15 patients transplanted within 6 months (group2). In all patients anthropometric, clinical and metabolic param-eters were evaluated before transplant. We evaluated 1) carotidstiffness (cPWV), intima-media thickness (cIMT), and presence ofplaques (CP), 2) echocardiographic parameters (E/A, thickness ofthe interventricular septum, left ventricular mass) and 3) visceraladiposity by epicardial fat thickness before and at 6 and 12 monthafter transplant.

RESULTS: All metabolic parameters (Cholesterol, triglycerides,fasting glucose), and blood pressure increased after OLT. Metabolicsyndrome were present in 19%, 57% and 54% of patients at time0, 6 and 12 months respectively (p = 0.04), while the preva-lence of diabetes did not significantly differ before and after OLT.A significant increase of carotid stiffness (7.2 ± 1,5 vs 8.3 ± 1.6,p = 0,05), and interventricular septum thickness (10.2 ± 0.4 vs12.2 ± 2.2, p = 0,02) and decrease of diastolic function (E/A 1.2 ± 0.4vs 0.86 ± 0.3, p < 0,01) was observed after 6 months from trans-plant. The cIMT and epicardial fat thickness significantly increasedafter 12 months (0.76 ± 0.2 vs 0.89 ± 0.2, p = 0.05) and (5.7 ± 3.2 vs8.0 ± 3.6, p = 0.05).

CONCLUSIONS: Metabolic Syndrome is highly prevalent in livertransplanted patients and cardiovascular alterations occur veryearly after liver transplantation. Recognition and promptly treat-
ment of these conditions may impact long- term survival of thesepatients.

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-30

CREENING FOR THE IDENTIFICATION OFUTOIMMUNE OR LYMPHOPROLIFERATIVENSET IN PATIENTS NAïVE TO HCV ANTIVIRALREATMENT

. Gulli 1, U. Basile 2, L. Colacicco 2, L. Miele 1,. De Matthaeis 1, P. Cattani 3, G.L. Rapaccini 1

Department of Internal Medicine, Istitute ofnternal Medicine and Geriatry- Catholic Universityf the Sacred Heart, Rome, ItalyDepartment of Laboratory Medicine, Institute ofiochemistry -Catholic University of the Sacredeart, Rome, ItalyDepartment of Laboratory Medicine, Institute oficrobiology -Catholic University of the Sacredeart, Rome, Italy

Introduction. Hepatitis C virus(HCV) may be responsible ofxtra-hepatic manifestations.A chronic infection of immunocom-etent cells is most likely at the origin of a benign mono-oligoclonallymphocyte proliferation, typically observed in mixed cryoglob-

linemia(10% showing late B-NHL).Aim. Identify early markers of autoimmune lymphoprolifera-

ive disease onset in a group of antiviral treatment-naïve patientsnfected by HCV that could identify the transition between a state ofilent autoimmune and lymphoproliferative conditions and frankisease.

Material, Methods and Results.Fourty patients wereecruited. Antinuclear antibodies(ANA) were detected by indirectmmunofluorescence. Autoantibody detection of IgG directedgainst M2,gp210,SP100,LKM1,LC1,SLA,Factin antigens were per-ormed by Immunodot analysis. Free light chain(FLC) detectionere carried out by turbidimetric assay. Cryoglobulin and cryofib-

inogen analysis was carried out following the guidelines of theIBIOC. Our results show an 84% prevalence of cryoglobulinemian samples collected from HCVinfected patients. Of these, 27%howed ANA positivity a negligible percentage of autoantibodyiver disease and absence of positivity of cryofibrinogen. The mostignificant result concerns the finding of high doses of FLC in3% of patients, of which 21% showing an abnormally elevated/l ratio. Statistical analysis suggests that patients presentingryoglobulinemia and FLCratio above 1.6 are also ANApositive.

Conclusions. ANA positivity is indicative of the presence of aersistent antigenic stimulus by the virus and the activation of anyutoimmune clones.The presence of cryoglobulinemia suggests aontinuous lymphocyte stimulation. Interestingly, our results sug-est a possible role for the presence of high levels of FLCs and theirse to identify the transition between a silent state of probableutoimmune lymphoproliferative disease or a frank illness, using/l ratio as a cut-off value. The presence of a subpopulation of HCV-ositive patients may open new scenarios to targeted therapeuticreatment strategies in subclinical phases. Our study is a contribu-
ion to presenting a panel of potential predictive markers of diseaserogression.

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F-31

THE FIRST STEP FOR THE PREVENTION OFASCITES IN PATIENTS WITH CIRRHOSIS: THEIDENTIFICATION OF AN EASILY ACCESSIBLEPREDICTOR OF THE FIRST ONSET OF ASCITES

S. Piano 1,2, M. Tonon 1,2, S. Fasolato 1,2,G. Bombonato 1, A. Romano 2, E. Gola 2,A. Brocca 2, F. Morando 2, M. Cavallin 2,P. Angeli 1,2

1 Unit of Hepatic Emergencies and LiverTransplantation, University of Padua, Padua, Italy2 Department of Medicine, University of Padua,Padua, Italy

Background and aim: Ascites represents the hallmark ofdecompensation in patients with cirrhosis. Apart from the assess-ment of HVPG, that is not yet part of clinical practice, predictorsof ascites onset are lacking, restricting the adoption of preventivestrategies. The presence of a modest layer of fluid (MLF) aroundthe liver and/or the spleen at ultrasound examination is a frequentfinding in patients with compensated cirrhosis. The meaning of MLFhas never been investigated in these patients. Thus, the aim of thisstudy was to evaluate if the MLF precedes the appearance of ascites.

Methods: 248 consecutive patients with cirrhosis withoutascites, were enrolled and followed up until death, liver transplan-tation and/or for a maximum of 10 years.

Results: Ninety-three patients (37.5%) had MLF at inclusion.Patients with MLF were older (58.3vs50.8 years; p < 0.001), hada higher MELD score (11.8vs10.7; p = 0.028), a higher Child-Pugh score (7.4vs6.4; < 0.001) and a lower mean arterial pressure(92.1vs97.4; p < 0.001) than patients without MLF. Ten-year prob-ability to develop overt ascites was significantly higher in patientswith MLF than without MLF (38.3vs22.7%;p < 0.001). MLF was foundto be an independent predictor of the development of over ascites(hazard ratio[HR] = 2.2; p = 0.023) as well as albumin (HR = 0.86;p < 0.001) and treatment with �-blockers (HR = 2.29; p = 0.024).

Conclusions: The presence of MLF and the use of �-blockerswithout a verfied effect on HVPG were found to be the strongestpredictors of ascites onset in patients with cirrhosis. MLF is an easilyaccessible predictor of the appearance of overt ascites in patientswith cirrhosis. Thus, it can be used to plan preventive interventions

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-32

HARACTERISTICS OF PATIENTS WITH CHRONICEPATITIS C WHO HAVE FAILED PASTREATMENT OR WHO HAVE NEVER BEENREATED: HOW MUCH ROOM LEFT FORNTERFERON?

. Ciaccio 1, S. Okolicsanyi 1, M. Rota 2,.A. Cortesi 3, S. De Salvia 1, M. Gemma 1,. Vinci 4, L.G. Mantovani 3, L.S. Belli 4,. Strazzabosco 1,5

Department of Surgery and Translational Medicine,niversity of Milano-Bicocca, Milan, ItalyDepartment of Health Sciences, Centre ofiostatistics for Clinical Epidemiology, University ofilano-Bicocca, Milan, ItalyResearch Centre on Public Health (CESP), Universityf Milano-Bicocca, Milan, ItalyDepartment of Hepatology and Gastroenterology,iver Unit, Niguarda Hospital, Milan, ItalyLiver Center & Section of Digestive Diseases,epartment of Internal Medicine, Yale Universitychool of Medicine, New Haven, CT, United States

Background/aims: According to current guidelines on manage-ent of Chronic Hepatitis C (CHC), both interferon (IFN)-containing

nd IFN-free treatment combinations may be offered to patients,epending on viral genotype, previous treatment and contraindi-ations to IFN. The past two decades of treatment have selected aarge population of patients ineligible to IFN. Our aim is to charac-erize the current CHC population in order to best drive resourcellocation.

Patients and methods: we retrospectively retrieved the char-cteristics and outcomes of a cohort of 801 consecutive CHCutpatients followed-up at two tertiary Centers in Northern Italy.aseline characteristics of never treated and treated CHC patientsere compared by means of Chi-Square test for categorical and

-test for continuous variables.Results: fifty-three percent (426 out of 801) of patients had pre-

iously received treatment. Among them, 103 (24%) had achievedVR; 323 (76%) had failed, of whom 74 (23%) did not completend 249 (77%) did not respond to treatment. Adverse events werehe main cause of treatment withdrawal (90%). Forty-seven per-ent (n = 375) of all participants had never received any treatment,ostly because of contraindications (27% overall, hematologic 11%,

sychiatric 24%, cardiovascular 15%, autoimmune 7%, others 43%),dvanced age (27%), personal choice (15%) or no fibrosis (10%).ever treated patients differed significantly (p < 0.01) from treatednes by age (61 vs 56 years) and gender (male 49 vs 65%). Six-undred and ninety-eight patients (323 failed plus 375 neverreated) are awaiting new antiviral therapy, of which 442 (63%) areandidate to all-oral combinations because of contraindications orast intolerance to IFN, while the remaining 256 (37%) may stilleceive an IFN-containing regimen.

Conclusions: 37% of the CHC patients being followed in our clin-cs may receive IFN-containing protocols with newer DAAs, while
bout the two thirds of these patients are obligated candidates forhe more costly, IFN-free regimens.

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F-33

ELF TEST IS A RELIABLE TOOL FORNON-INVASIVE DIAGNOSIS OF LIVER FIBROSIS INPATIENTS WITH NONALCOHOLIC FATTY LIVERDISEASE

L. Miele 1, M.A. Isgrò 2, T. De Michele 2,G. Marrone 1, C. Cefalo 1, S. Racco 1, L. Viti 1,A. Giannace 2, C. Morlacchi 2, G.L. Rapaccini 1,A. Gasbarrini 1, C. Zuppi 2, A. Grieco 1

1 Department of Internal Medicine, CatholicUniversity of Rome, Rome, Italy2 Department of Laboratory Medicine, CatholicUniversity of Rome, Rome, Italy

Introduction: Recently, the serum Enhanced Liver Fibrosis (ELF)Test has been developed for staging liver fibrosis in patients withchronic liver diseases.

Aim: The aim of our study was to evaluate the ELF Test perfor-mance in predicting fibrosis stage in an independent adult cohortof NAFLD patients.

Materials and methods: 82 patients (mean age 46 years)with suspected NAFLD were enrolled undergoing percuta-neous liver biopsy and serum sampling. Fibrosis was assessedand scored by using the modified Brunt classification (F0 = nofibrosis; F1 = perisinusoidal/periportal; F2 = perisinusoidal and por-tal/periportal; F3 = bridging fibrosis; F4 = cirrhosis). The ELF testwas determined in all patients by means of an algorithm combininghyaluronic acid, amino-terminal propeptide of type III collagen andtissue inhibitor of metalloproteinase 1. Diagnostic accuracy wasassessed determining the area under receiver operating character-istic curves (AUCs).

Results: The distribution of fibrosis stages in our cohort wasas follows: F0 = 7.3% (n = 6), F1 = 39.0% (n = 32), F2 = 35.4% (n = 29),F3 = 6.1% (n = 5), F4 = 12.2% (n = 10). The ELF Test had an AUC of 0.988(95% Confidence Interval C.I. 0.967-1.008; P < 0.001) for distinguish-ing cirrhosis, 0.948 (C.I. 0.883-1.014; P < 0.001) for severe fibrosis,0.682 (C.I. 0.568-0.797; P = 0.005) for significant fibrosis and 0.658(C.I. 0.401-0.915; P = 0.200) for any fibrosis. ELF scores were signifi-cantly higher in patients with severe fibrosis/cirrhosis in respectto ones with no/mild/moderate fibrosis (median 11.26 vs. 8.53;P < 0.001). Severe fibrosis and cirrhosis were correctly identified in91% of patients.

Conclusions: In our cohort of NAFLD patients, the ELF test wasable to discriminate severe fibrosis and cirrhosis with an excellentdiagnostic accuracy. It may result useful for the selection of cases

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-34

MPACT OF ANTICOAGULATION ON THEVOLUTION OF PORTAL VEIN THROMBOSIS INIRRHOSIS: A RETROSPECTIVE ANALYSIS

. Pettinari 1, S. Landi 1, S. Berardi 2, C. Serra 2,. Pianta 2, P. Caraceni 3, G. Verucchi 4,. Cescon 5, M.R. Tamè 6, L. Bolondi 1,

. Piscaglia 1

Unit of Internal Medicine, Department of Digestiveisease, General and University Hospital.Orsola-Malpighi, Bologna, ItalyUnit of Internal Medicine and Organs Failure,eneral and University Hospital S.Orsola-Malpighi,ologna, ItalyUnit of Semeiotica, Department of Digestiveisease, General and University Hospital.Orsola-Malpighi, Bologna, ItalyUnit of Infectious Disease, General and Universityospital S.Orsola-Malpighi, Bologna, ItalyUnit of Hepatobiliary Surgery, Department ofrgans Failure and Transplantations, General andniversity Hospital S.Orsola-Malpighi, Bologna, ItalyUnit of Gastroenterology, Department of Digestiveisease, General and University Hospital.Orsola-Malpighi, Bologna, Italy

Background: Non-neoplastic portal vein thrombosis (PVT) iscommon complication in cirrhosis, however no consensus exist

bout any management and therapeutical algorithm for this con-ition.

Aim of our study was to investigate the evolution of PVT inirrhosis and the influence of anticoagulant treatment.

Materials and methods: A retrospective analysis of 68 cirrhoticatients with non-malignant PVT observed at the S.Orsola-alpighi General and University Hospital from January 2008 toecember 2013 was performed. Patients with HCC were notxcluded provided that PVT was ascertained as non neoplastic.hrombosis diagnosis and assessment of recanalization were per-ormed using Doppler ultrasound, spiral CT or MRI. Anticoagulantsere administered to only 25 of 68 patients at thrombosis detec-

ion. Median follow- up was 24 months in treated and 17 monthsn untreated patients.

Results: In the 43 patients not receiving anticoagulant ther-py, PVT remained stable in 21 (48.8%), progressed in 15 (34.8%), 8f whom subsequently started anticoagulants and spontaneouslymproved in 7 patients (16.4%). Recanalization was achieved in 16 of5 patients (64%) receving anticoagulation (11 complete and 5 par-ial PVT), while no recanalization was observed in 9 anticoagulatedatients (36%). Seven of 11 patients who had achieved completeecanalization suffered PVT recurrence after stopping anticoagula-ion. Bleeding, but not lethal complications occurred in 7 treatedatients (28%), of which 3 were suspected to be anticoagulation-elated.

Conclusion: Our study confirms that anticoagulation ther-py influences the course of PVT, favoring recanalization andreventing thrombus extension. However discontinuation of anti-
oagulation therapy is associated with high risk of recurrenthrombosis. Anticoagulant treatment appeared to be relatively safe.

sease 47S (2015) e43–e66 e59

F-35

SERUM SCCA-IGM CAN PREDICT THEDEVELOPMENT OF HCC IN PATIENTS WITHHCV-CIRRHOSIS

G. Pieri 1, S. Marenco 1, F. Lantieri 2,I. Baldissarro 1, L. Bruzzone 1, V. Fazio 1,S. Labanca 1, G. Sammito 1, V. Savarino 1,A. Picciotto 1

1 U.O.s. Diagnosi e Terapia delle Epatopatie, IRCCS,San Martino IST, Genova, Italy2 DISSAL, Biostatistics Dep., University of Genoa, Italy

Introduction and aim. The serum level of circulating squa-mous cellular carcinoma antigen (SCCA)-IgM immunocomplex hasbeen proposed as marker for diagnosis of hepatocellular carcinoma(HCC) and its progressive increase is associated with developmentof tumor. We studied the role of a single SCCA-IgM measurementin predicting the development of HCC.

Material and Methods. We retrospectively selected 93 patientswith cirrhosis (26 with HCC and 67 without HCC at enrollment). Foreach patient, we chose serum samples collected at least 6 monthsbefore the diagnosis of HCC or at least 12 months before the enrol-ment. We measured SCCA-IgM using an ELISA assay kit. We appliedthe ROC-curve for the best cut-off; independent Student’s t-testand Fisher’s test for parametric data; Mann-Whitney U-test fornon-parametric data; Kaplan-Meier to test the survival.

Results. There were no differences for age, gender and HCV-infection between the groups with and without HCC. The bestSCCA-IgM predictive value of HCC development was 192 AU/mL(sensibility 65.4%, specificity 62.7%, AUROC 0.607). SCCA-IgM lev-els were higher in HCV versus non-HCV patients (p = 0.027), bothin HCC and HCC-free group. Considering the HCC occurrence andthe HCV etiology together (ANOVA), the second one was betterrelated to SCCA-IgM (p = 0.04). SCCA-IgM levels > 192 AU/mL weremore frequent in patients who developed HCC than in pantientswho did not develop HCC (p = 0.032). The predictivity of SCCA-IgM levels increased considering male HCV-positive patients alone(sensitivity 85.7%, specificity 57.1%, AUC 0.707) with the same cut-off. The survival analysis revealed that the mean HCC-free survivalin patients with SCCA-IgM <192 AU/mL was longer than in patientswith SCCA-IgM 3192 AU/mL (105.1 vs 84.7 months, p = 0.024).

Conclusions. SCCA-IgM levels are higher in HCV-positivepatients and can help identifying those with higher risk of earlierdevelopment of HCC, using a cut-off of 192 AU/mL.


F-36

ANTIHBS IS A USEFUL TOOL FOR SEARCHING THERISK OF HEPATITIS B REATIVATION: AMETA-ANALYSIS OF OBSERVATIONAL STUDIESIN PATIENTS WITH LYMPHOMA

M. Fiore 1, I. Baldi 2, A. Floreani 3

1 Infectious Disease Unit, University Hospital ofTrieste, Trieste, Italy2 Department of Cardiac, Thoracic and VascularSciences, University of Padua, Padua, Italy3 Department of Surgical, Oncology andGastroenterology, University of Padua, Padua, Italy

Background: Reactivation of HBV is a well-recognized com-plication following immunosuppression in patients with pastinfection. The International guidelines (AASLD, EASL) recommend

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etection of antiHBc regardless of antiHBs status in subjects under-oing chemotherapy, and prophylaxis in those with viral loadetectable.

Aim: To perform a meta-analysis of papers dealing on thisssue searching for two primary outcome measures: i) the propor-ion of reactivation of HBV in HBsAg-ve patients according to thentiHBc status; ii) the proportion of reactivation of HBV in HBsAg-e/antiHBc + ve patients according to the antiHBs status.

Methods: We searched the PuMed collection from any date toebruary 28, 2014 for the following terms: “HBV, hepatitis B, reac-ive. Of the 1,226 screened papers only 11 papers fulfilling thenclusion criteria were included (full papers). The Der Simoniannd Laird effects models together with the Duval and Tweedie nonarametric “trim and fill” method of accounting for publication biasere used.

Results: Patients positive for antiHBc alone showed a 4%ncreased risk of reactivation of HBV which was significantly higherhan antiHBc-ve subjects (95%CI: 1-6%, p = .002). The asymmetry ofunnel plot analysis suggested the presence of a publication bias,hus the risk differences were reduced to 2.3% (p = .09) according tohe “trim and fill” analysis. Considering the second endpoint mea-ure, a significant reduction of the risk of reactivation was obtainedooling antiHBs + ve and antiHBs-neg subjects (-8%, 95%CI:-12 to4%, p < .001). Finally, based on these data, an algorithm for HBVrophylaxis was constructed, recommending antiHBs searching asuseful tool for management of HBV reactivation.

Conclusion: The antiHBs assay may reduce the risk for HBVeactivation and HBV-associated morbidity/mortality in patientsith past HBV infection undergoing chemotherapy for lymphoma.


-37

EARLY” SUBCLINICAL LEFT VENTRICULARISFUNCTION IN PATIENTS WITH HEPATITIS CIRUS INFECTION

. Rocco 2, M. Sanduzzi Zamparelli 2,

. Angrisani 2, D. Compare 2, C. Santoro 2,. Galderisi 1, G. Nardone 1

Hypertension Research Center and Department ofedical Translational Sciences, Federico II University

f Naples, Naples, ItalyDepartment of Clinical Medicine and Surgery,astroenterology Unit, Federico II University ofaples, Naples, Italy

Introduction: Hepatitis C virus (HCV)infection has been associ-ted with several extrahepatic manifestations in a variety of tissuesnd organs, including the myocardium. Reduced myocardial per-usion or increased pro-B-type natriuretic peptide (NT-proBNP)ave been described in HCV-infected individuals but, until now,o study clearly demonstrated the occurrence of myocardial dys-

unction in the early stages of HCV-related liver diseases.Aim: to evaluate the impact of HCV infection on left ventricular

LV) geometry and function by a standard 2D and 3D echocardiog-aphy.

Material and Methods: Thirty consecutive patients with histo-ogically proved HCV-related chronic hepatitis and twenty normalontrols (NCs), comparable for age and gender prevalence werenrolled in the study. Exclusion criteria were: other causes ofiver diseases, coronary artery and/or valvular heart disease, heart
ailure, cardiomyopathies, atrial fibrillation or alcohol abuse. Thechocardiography protocol consisted of: standard echo Dopplerlus LV volumetric and systolic function analysis through 3D
sease 47S (2015) e43–e66

echocardiography. Left ventricular systolic function thorough 3Decho was evaluated by STE derived global longitudinal strain (GLS),global circumferential strain (GCS), global area strain (GAS) andglobal radial strain (GRS).

Results: Overall we enrolled 16 males and 14 females with HCV(genotype 1) related chronic liver disease. Mean age was 60.9 ± 6.5yrs. NCs were comparable for age, gender distribution, body massindex, heart rate and blood pressure. In HCV infected subjectsstandard 2D echocardiography point out a transmitral E/A ratiomarginally lower (0.84 ± 0.2) compared to NC (1.01 ± 0.3) (p = 0.04).Moreover, more detail 3D STE showed a subclinical alteration ofLV function documented by lower GCS (HCV = -15.3 ± 2.2%, NC = -17.6 ± 3.9%, p = 0.02), GAS (-25.2 ± 2.6%, NC -29.9 ± 5.2%, p = 0.03),and GRS (37.6 ± 4.6% vs 44.9 ± 12%, p = 0.04) whereas HCV-releatedchanges of GLS were not significant.

Conclusion: HCV chronic infection is associated a subclinicalmyocarditis-like damage documented by the alteration of the cir-cumferential fibers of the ventricular mid wall.


F-38

OXIDATIVE STRESS, MITOCHONDRIA DAMAGEAND MATRIX METALLOPROTEASE ACTIVATIONIN THE PATHOGENESIS OF NAFLD

C. Berardo 1,∗, G. Palladini 1, L.G. Di Pasqua 1,V. Rizzo 2, S. Perlini 1, P. Richelmi 1, M. Vairetti 1,A. Ferrigno 1

1 Department of Internal Medicine and Therapeutics,Fondazione IRCCS Policlinico S. Matteo, University ofPavia, Pavia, Italy2 Department of Molecular Medicine, FondazioneIRCCS Policlinico S. Matteo, University of Pavia,Pavia, Italy

Background and Aims: Animal models of hepatic steatosishave allowed the understanding of mechanisms involved in thepathogenesis of non-alcholic fatty liver disease (NAFLD). By com-paring these models a working hypothesis for NAFLD pathogenesisemerges as well as the mechanism implicated in the progress tononalcoholic steatohepatitis (NASH). This study investigated theoxidative stress, mitochondria damage and matrix metalloproteaseactivation in fatty livers using two rat models of NAFLD such as themethionine and choline deficient (MCD) diet model and obese fa/farats.

Methods. Male Wistar rats underwent to NAFLD induced by 4-week MCD diet; blood samples and hepatic biopsies were collectedup to 4 weeks. 12-week male old obese (fa/fa) and lean (fa/-) maleZucker rats were also used. Serum hepatic enzymes (AST, ALT, LDHand g-GT) and total and direct bilirubin were quantified. Tissueglutathione (GSH), lipid peroxides, ATP/ADP ratio and matrix met-alloproteinase activation (MMP-2, MMP-9) were also evaluated.

Results: No significant changes in serum AST, ALT, LDH and g-GT were found comparing the two animal models. A marked serumincrease in total and direct bilirubin at IV weeks in MCD rats wasdetected. Mostly decrease in tissue GSH levels and increase in thelipid peroxides were found in the MCD group. A better mitochon-drial function as documented by the ATP/ADP ratio and no MMP-9and poor MMP-2 activation was measured in obese fa/fa rats whencompared with the MCD animals. Instead, in MCD rats a significantincrease of MMP2 activation occurred.

Conclusions: MCD rats exhibit a marked oxidative stress andmitochondria damage concomitant with an MMP-2 activation. Thereported spontaneous development to severe NASH observed only

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n MCD rats and not in obese fa/fa animals might be associated tohese events already detectable in the early period of treatment.

Funding: Supported by Fondazione Cariplo, grant n◦ 2011-0439.


-39

BSAG NEGATIVE ANTI-HBC POSITIVEEROLOGY INCREASES THE RISK OF HCC INHRONIC HEPATITIS PATIENTS: AETA-ANALYSIS

. Coppola 1, L. Onorato 1, C. Sagnelli 2,. Sagnelli 1, I.F. Angelillo 3

Department of Mental Health and Public Medicine,ection of Infectious Diseases, Second University ofaples, Naples, ItalyDepartment of Clinical and Experimental Medicinend Surgery, Second University of Naples, Naples,talyDepartment of Experimental Medicine, Secondniversity of Naples, Naples, Italy

Background and Aims: We performed a meta-analysis to ascer-ain if patients with chronic hepatitis (CH) of different etiologynd anti-HBc positive are at a higher risk to develop hepatocellulararcinoma (HCC) than those anti-HBc negative.

Methods: studies meting these criteria were included:nvestigating on the relationship between HBsAg-negative/anti-Bc-positive serology and occurrence of HCC, being a case-controlr cohort study, providing relative risk or odds ratios and 95% confi-ence intervals, being available as full text written in English, beingublished and indexed up to September 2014.

Results: Twenty-six studies met inclusion criteria, allowing aeta-analysis on 14,558 patients. Table 1 shows the results of theeta-analysisConclusions: This meta-analysis shows that in HBsAg negative

H patients anti-HBc positivity is strongly associated with HCC, anssociation observed in all subgroups established by the stage of
he disease, aetiology and ethnicity

able 1eta-analysis data on the development of HCC in CH anti-HBc positive or anti-HBc negat

HCC in No of studies N◦ of patientsAnti-HBc + /anti-HBc-

N◦ and (%) HAnti-HBc+/a

All subjects 26 5581/8977 1705 (30.5)chronic liver diseases* 23 4614/5034 1513 (32.8)Chronic hepatitis 3 370/408 76 (20.5)/62Liver cirrhosis 7 973/959 320 (32.9)/1Asian subjects 16 3639/4230 1231 (33.8)Non-Asian subjects 10 1942/4747 474 (24.4)/3Anti-HCV positive subjects 20 4747/8082 1348 (28.4)Anti-HBs positiveˆ 9 935/4198 223 (23.8)/3Anti-HBs negativeˆˆ 9 719/4198 331 (46.0)/3Anti-HBs positiveˆˆ 9 935/719 223 (23.8)/3

Excluding the healthy controls. ˆ Anti-HBs/anti-HBc positive subjects vs. anti-HBs/antBs/anti-HBc negative subjects. ˆˆˆ Anti-HBs/anti-HBc positive subjects vs. anti-HBs nega

sease 47S (2015) e43–e66 e61

F-40

A STRATEGY TO FAVOR THE ACCESS OFIRREGULAR AND REFUGEE MIGRANTS TO ASCREENING PROGRAM FOR HBV, HCV AND HIVINFECTION

N. Coppola 1, L. Alessio 1,2, L. Gualdieri 3,M. Pisaturo 4,5, C. Sagnelli 2,6, N. Caprio 1,7,R. Maffei 1,3, M. Starace 1, I.F. Angelillo 8,G. Pasquale 1, E. Sagnelli 5

1 Department of Mental Health and Public Medicine,Section of Infectious Diseases, Second University ofNaples, Naples, Italy2 Medical Center, Centro Sociale ex Canapificio,Caserta, Italy3 Medical Center, Centro per la Tutela della Salutedegli Immigrati, Naples, Italy4 Medical Center, Centro di Accoglienza “La tenda diAbramo”, Caserta, Italy5 Infectious Diseases Unit, AORN Sant’Anna e SanSebastiano, Caserta, Italy6 Department of Clinical and Experimental Medicineand Surgery, Second University of Naples, Naples,Italy7 Medical center, Centro Suore Missionarie dellaCarità, Naples, Italy8 Department of Experimental Medicine, SecondUniversity of Naples, Naples, Italy

Background: Half immigrants are irregular or refugee fre-quently excluded from social life by linguistic, cultural andsocioeconomic barriers also limiting their access to the healthcareservices.

Aim: To analyze the use of strategies favoring the access ofirregular or refugee migrants to a screening program for HBV, HCVand HIV infection.

Methods: A screening for HBV, HCV or HIV infection was pro-posed to 926 migrants by a team (doctors, nurses and cultural

ive.

CCnti-HBc-

RR (efficacy) 95% CI (efficacy) p Heterogeneity test(Q;p;I2,%)

/1257 (14.0) 1.58 1.49-1.69 0.000 99.91;0.000;75.0/1119 (22.2) 1.45 1.36-1.54 0.000 101.62;0.000;75.4

(15.2) 1.44 1.18-1.77 0.000 1.16;0.56;0.096 (20.4) 1.30 1.12-1.52 0.001 10.85;0.093;44.7

/859 (20.3) 1.62 1.50-1.74 0.000 34.51;0.003;56.598 (8.4) 1.51 1.34-1.69 0.000 65.14;0.000;86.2

/999 (12.4) 1.52 1.4-1.63 0.000 64.62;0.000;72.184 (9.1) 1.17 1.01-1.35 0.03 43.78;0.000;81.784 (9.1) 1.75 1.56-1.96 0.000 120.61;0.000;93.431 (46.0) 0.63 0.55-0.72 0.000 18.61;0.017;57.0

i-HBc negative subjects. ˆˆ Anti-HBs negative/anti-HBc positive subjects vs. anti-tive/anti-HBc positive patients.

screening, free of charge and with no bureaucracy, was accepted by882 (95.2%) migrants, 625 irregular and 257 refugee, and performed

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Results: 78 (8.7%) migrants were HBsAg positive, 35 (3.7%)nti-HCV positive, 11 (1.3%) anti-HIV positive and 7 (0.8%) had aultiple infection. Of the 801 HBsAg negative patients, 373 (40.2%)ere anti-HBc positive. All migrants with a detectable marker ofBV, HCV or HIV infection were unaware of their serological sta-

us. The HBsAg positivity rate was high (13.9%) in migrants fromub-Saharan Africa and intermediate in those from eastern Europe6.1% of 194 cases), northern Africa (2.5% of 80 cases) and Indian-akistani area (3.2% of 126 cases). Anti-HCV was more frequentn migrants from eastern Europe (6.2%) and Indian-Pakistani areas7%) than in those from sub-Saharan (3.8%) and northern Africa2.5%). Anti-HIV was detected in 17 (3.8%) migrants from sub-aharan Africa, in 5 (2.4%) from eastern-Europe, in 1 (0.8%) fromndian-Pakistani area and in none from northern Africa.

Discussion. This investigation identified 131 migrants withBV, HCV or HIV infection unaware of their serological status, sug-esting the need of a strong action of Healthcare Authorities of Italy,ay be using our model, to favor the access of migrant populations

o healthcare services and to support screening and educationalrograms


-41

AMIVUDINE PROPHYLAXIS PREVENTSEPATITIS B REACTIVATION INBSAG-NEGATIVE/ANTI-HBC-POSITIVEATIENTS UNDERGOING RITUXIMAB-BASEDHEMOTHERAPY FOR NON–HODGKIN’S B CELLYMPHOMA

. Viganò 1, G. Grossi 2, E. Borsotti 2,

. Cappelletti 1, M. Goldaniga 3, L. Farina 4,

. Rumi 1, P. Corradini 4, L. Baldini 3,

. Colombo 2, P. Lampertico 2

Division of Hepatology, Ospedale San Giuseppe,niversity of Milan, Milan, ItalyDivision of Gastroenterology and Hepatology,ondazione IRCCS Cà Granda Ospedale Maggioreoliclinico, University of Milan, Milan, ItalyDepartment of Oncohematology, Fondazione IRCCSà Granda Ospedale Maggiore Policlinico, Universityf Milan, Milan, ItalyDivision of Hematology, IRCCS Istituto Nazionaleumori, University of Milan, Milan, Italy

Introduction and Aim: Hepatitis B surface antigen (HBsAg)-egative/anti-hepatitis B core antigen (anti-HBc)-positive patientsndergoing Rituximab (RTX)-based chemotherapy (R-CT) foron–Hodgkin’s B cell lymphoma (NHL) face up to 27% risk of hep-titis B virus (HBV) reactivation. Aim of the study was to assesshe efficacy and safety of lamivudine (LMV) prophylaxis in theseatients.

Materials and Methods, Results: Sixty-seven consecutiveBsAg-negative/anti-HBc-positive patients (median age 70 yrs,1% males, 13% HCV seropositive, 100% serum HBV DNA negativey sensitive PCR assay, 75% anti-HBs positive, 100% with ALT < ULN)ndergoing different R-CT protocols (48% R-CHOP) in two hemato-

ogical centres were retrospectively enrolled. LMV prophylaxis wastarted before the first R-CT dose and planned to last for 18 months
fter the end of R-CT. R-CT was administered for 6 cycles (range: 1-5) during 5 (range: 1-38) months, while LMV was administered for0 (range: 3-54) months. Serum ALT, HBsAg and HBV DNA levels by
sease 47S (2015) e43–e66

PCR assay were assessed every 3-4 months. All patients remainedHBV DNA and HBsAg negative during a median of 20 months (range:2-60), including both the LMV prophylaxis and the subsequent offtreatment follow-up. Anti-HBs titers declined in 18% of patientsbut none lost serum reactivity for anti-HBs. Overall, 2 patients hadan increase of ALT (> ULN) but this event was unrelated to HBVreactivation and 7 patients died of liver unrelated causes. No safetyissues related to LMV were recorded.

Conclusions: LMV monotherapy is an inexpensive, safe andeffective prophylaxis regimen to prevent HBV reactivation in such ahigh-risk population as HBsAg-negative/anti-HBc-positive patientsreceiving RTX-based chemotherapy for non–Hodgkin’s B cell lym-phoma.


F-42

HBV GENETIC COMPARTIMENTALIZATION,VARIABILITY AND MOLECULAR CORRELATES OFHISTOLOGIC AND IMMUNOHISTOCHEMICALASPECTS IN LIVER TISSUE: IMPLICATIONS FORTHE CLINICAL MANAGEMENT OF PATIENTSWITH CHRONIC HEPATITIS B

C. Minosse 1, A. Baiocchini 2, M. Selleri 1,E. Giombini 1, S. Coen 1, P. Zaccaro 1, G. Rozera 1,D. Vincenti 1, F. Del Nonno 2, U. Visco Comandini 3,R. Lionetti 3, M. Montalbano 3, G. D’Offizi 3,M. Vivarelli 4, M.R. Capobianchi 1, S. Menzo 5

1 Laboratory of Virology, National Institute forInfectious Diseases “L. Spallanzani”, Rome, Italy2 Laboratoy of Pathology, National Institute forInfectious Diseases “L. Spallanzani”, Rome, Italy3 1st Clinical Division, National Institute forInfectious Diseases “L. Spallanzani”, Rome, Italy4 Hepatobiliary and Transplantation Surgery Unit,Università Politecnica delle Marche, Ancona, Italy5 Laboratory of Virology, Università Politecnica delleMarche, Ancona, Italy

Introduction In chronic hepatitis B (CHB), viral intrahepaticmolecular (ccc- and total DNA) and immunohisto chemical mark-ers should allow the best representation of viral biology and organpathology. Presently, the role of these intrahepatic markers in thepathogenesis of CHB and their clinical significance are still unclear.Furthermore, the the genetic determinants of intrahepatic HBV(compared to plasma), have essentially never been considered.

Aim This study focused on the integrated analysis of intra-hepatic molecular and histochemical markers to elucidate somepathogenetic mechanisms underlying chronic HBV infection anddisease.

Materials and Methods, Results The study enrolled 35 consec-utive untreated CHB patients. In addition to classical virological andclinical parameters, intrahepatic cccDNA, total DNA quantification(by real-time PCR) and intrahepatic viral antigen distribution (byimmunohistochemistry) were evaluated in liver tissue. Conven-tional and next generation sequencing of the pre-core and pre-sregions of viral genome was performed on all viral compartments.

Precore mutations were shown to be responsible of cytoplas-matic accumulation of HBsAg and reduced viral replicative capacity(totalDNA/cccDNA). A missense precore deletion was associatedto G3 steatosis in one patient. Mainstream intrahepatic virus was
absent from plasma in 2 patients. Other patients displayed moder-ate compartimentalization. Interestingly, variants not appearing inthe peripheral blood carried missense indels in the precore region,

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riving the synthesis of abnormal peptides, possibly leading toirion assembly defects and to the intracytoplasmatic accumula-ion of HBsAg.

Conclusions Selective pressure by the immune system selectsariants in the precore region that are not proportionally detectablen the periphery, possibly due to the abnormal release of defectiveirions. This may also have clinical implications for hepatic pathol-gy (steatosis) and may lead to the underestimation of viral load. Aetter comprehension of the relationship between intrahepatic anderipheral (virological and clinical) markers may confer to these aigher diagnostic power.


-43

ARCOPENIA AT FIRST DIAGNOSIS PREDICTS AEDUCED SURVIVAL IN PATIENTS AFFECTED BYEPATOCARCINOMA

. Antonelli 1, P. Begini 1, E. Gigante 1,. Carbonetti 2, E. Iannicelli 2, S. Gallina 1,. Pellicelli 3, L. Miglioresi 3, G. Delle Fave 1,. Marignani 1

Digestive and Liver Diseases Dpt., Sant Andreaospital, School of Medicine and Psychology,apienza University, Rome, ItalyRadiology Dpt., Saint Andrew Hospital, School ofedicine and Psychology, Sapienza University,

ome, ItalyLiver Unit., San Camillo Forlanini Hospital, Rome,

taly

Introduction Sarcopenia is a frequent complication and anndependent risk factor for mortality and clinical outcomes inatients (pts) with liver cirrhosis and in a variety of other clinicalonditions.

Aim of the study was to determine the prevalence of sarcopenian a cohort of cirrhotic pts at the first diagnosis of hepatocellulararcinoma (HCC) treated in a tertiary center and its influence onurvival.

Methods All consecutive HCC pts treated at our outpatient clinicyears 2004-2014) undergoing abdominal computed tomographyCT) were retrospectively studied with a software analyzing theross-sectional areas of muscles at L3 level. Data was normalizedor height obtaining the Skeletal Muscle Index (SMI) to measurearcopenia. Presence of sarcopenia was defined when SMI was41 cm2/m2 for women and ≤53cm2/m2 for men with a body mass

ndex (BMI) ≥25, and ≤43 cm2/m2 for men with BMI < 25.Results 92 HCC pts were evaluated [27F (29.4%), 65 M (70,6%].

ge at diagnosis was 71,9 years (30,7-86,4), while BMI was 24,717,5-36,7). Viral infection was the cause of liver disease in mostases. Distribution by CHILD score was as follows: A = 55,4%;= (42,4%); C = (2,2%). Median MELD score was 10 (6-18). Metastaticisease was present in 12% of cases. The distribution by BCLC was asollows: A = (41,3%);B = (25%), C = 28,3%, D = 5,4%. Overall, sarcope-ia was present in 40,2%. Baseline features were similar betweenarcopenic vs non sarcopenic pts. Sarcopenic pts were prevalentlyemales (p = 0,0044) and had a reduced mean Overall Survival of23 (95% C.I. 98 to 150) vs 66 (95% C.I. 47 to 84) weeks (p = 0,001).ultivariate analysis has shown that sarcopenia was indipendent

f age (p = 0,0027).Conclusions The prevalence of Sarcopenia in HCC pts is high,

ith a greater frequency in female pts. Presence of Sarcopenia atiagnosis of HCC is an important predictor of a reduced survival.


sease 47S (2015) e43–e66 e63

F-44

PANTOPRAZOLE AND RABEPRAZOLE DO NOTIMPACT THE ACTIVITY OF CYTOCHROME P450ASSESSED BY [13 C]-AMINOPYRINE BREATHTEST IN PATIENTS WITH LIVER CIRRHOSIS

A. Rocco 2, D. Angrisani 2, C. Rubicondo 2,L. Staiano 1, D. Amoruso 1, D. Compare 2,C. Coppola 1, G. Nardone 2

1 Hepatology and Interventional Ultrasound Unit,Gragnano Hospital, Gragnano, Italy2 Department of Clinical Medicine and Surgery,Gastroenterology Unit, Federico II University ofNaples, Italy

Introduction: Proton pump inhibitors (PPIs) are one of the mostwidely used drugs worldwide. Almost all PPIs undergo extensivehepatic metabolism via cytochrome (CYP)-P450 system, specifi-cally CYP-2C19 and CYP-3A4. Pantoprazole is metabolized throughthe CYP-P450 system, while the primary pathway of rabeprazolemetabolism is non enzymatic. In advanced liver diseases the activ-ity of drug metabolizing enzymes is impaired, thus increasingthe risk of drug accumulation, drug-drug interaction and adverseevents.13C-aminopyrine breath test (13C-ABT) is a noninvasive,liver function test that explores CYP enzyme activity.

Aim: to evaluate the effects of different PPIs on the activity ofCYPs by 13C-ABT in patients with Hepatitis C virus (HCV)-relatedliver cirrhosis.

Material and Methods: Thirty consecutive genotype 1 HCV-related liver cirrhosis patients, Child-Pugh A, were randomlyassigned to pantoprazole (40 mg/day) or rabeprazole (20 mg/day).

13C-ABT was performed before and 15 days after starting PPIby collecting breath samples baseline and at 30-minute inter-vals for 2 hours after oral administration of 13C-aminopyrine(2 mg/Kg body weight). 13C-enrichment of CO2 was determinedby purification-isotope ratio mass spectrometer. Results wereexpressed as maximum percentage of 13CO2-recovery per hour(max 13C% dose/h) at any time (“excretion peak”) and percent-age of 13CO2-cumulative dose recovered in 2 h (%13C cum dose at120 min).

Results: Overall, we enrolled 13 males and 17 females. Meanage was 60.9 ± 7.5 yrs. Age, gender distribution, BMI and labora-tory findings did not significantly differ among pantoprazole andrabeprazole group. Baseline, 13C-ABT results were altered in 13/30(43%) patients (6 in pantoprazole and 7 in rabeprazole group). Fif-teen days after PPIs, 13C-ABT did not significantly changed in termsof both %dose/h and %dose/cumulative, irrespective of PPI used.

Conclusion: Pantoprazole and rabeprazole do not significantlyaffect liver function in patients with HCV-related liver cirrhosis.

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-45

RANSIENT ELASTOGRAPHY IN PATIENTS WITHHRONIC AUTOIMMUNE HEPATITIS: IS A GOODOOL IN EVALUATION OF HEPATIC FIBROSIS?

. Onali 1,5, S. Cappellini 1,5, G. Serra 1,5,

. Zolfino 2,5, D. Murgia 3,5, M.L. Ponti 3,5,

. Balestrieri 1,5, M. Conti 1,5, O. Sorbello 4,5,

. Civolani 4,5, M. Casale 1,5, S. Casu 1,5,. Figorilli 1,5, M.C. Pasetto 1,5, A. Gaspardini 1,5,. Secci 1,5, C. Salustro 1,5, R. Ganga 3,5,.R. Piras 2,5, L. Demelia 4,5, L. Chessa 1,5

Center for the Study of Liver Diseases, Departmentf Medical Sciences “M.Aresu”, University of Cagliari,talyS.C. Gastroenterologia, AOB, Cagliari, ItalyS.C. Medicina Prima, AOB, Cagliari ItalyU.O. Gastroenterologia, AOU, Cagliari, ItalyAssociazione Epatologi Sardi

Background and aims Liver stiffness measurement (LSM) byransient elastography (TE, FibroScan) is a recently validated

ethod for noninvasive evaluation of fibrosis in chronic liver dis-ases, especially in patients with chronic hepatitis C.

The role of TE in evaluating liver fibrosis in chronic autoimmuneepatitis (AIH) patients has not been yet extensively investigated.herefore, we aimed to assess the diagnostic performance of TE inIH patients by comparison with other nonivasive methods, suchs APRI and FIB-4 score.

Methods Sixty four patients with AIH were consecutevelynrolled between January 2012 and August 2014. All patients weren treatment with steroid and or other immunosoppessive drugs,uch as azathioprine. TE was performed in all patients and livertiffness value was compared with the histological stage (Metavir)sing the Spearman correlation. APRI and FIB-4 score were alsoalculated. Patients who underwent to liver biopsy more than fiveears before the LSM were excluded. Patient with a clinical diag-osis of cirrhosis and no signs of portal hypertension were also

ncluded.Results 22 patients were analysed: M/F:3/19; mean age 48

ears; mean BMI 17 Kg/m2; mean AST 23UI/l (13-50); mean ALT8 UI/l (13-100); most of the patients had mild-moderate fibrosis15/23). Mean liver stiffness value was 6,4 Kpa (3-18).

The Spearman test showed a significant correlation betweenSM and histological stage (r = 0,56; p = 0,007). Mean liver stiffnessalue in cirrhotic patients was 10,6 Kpa vs 4,8 Kpa in non cirrhoticatients (p < 0,001). A significant correlation was also observedetween FIB-4 score and histological stage (r = 0,62, p = 0,002),lthough no differences were found between cirrhotic and nonirrhotic value (p = 0,56).

Conclusions Transient elastography could be an effective
ethod to assess and monitor hepatic fibrosis in chronic autoim-une hepatitis, especially in patients with advanced fibrosis.

sease 47S (2015) e43–e66

F-46

LITAF ROLE IN IN VIVO HEPATOCYTEPROLIFERATION AND IN HEPATOCARCINOMACELLS

N. Panera 1, A. Crudele 1, S. Ceccarelli 1, D. Gnani 1,C. De Stefanis 1, V. Nobili 1, A. Alisi 1

1 Liver Research Unit, Bambino Gesù ChildrenHospital and IRCCS, Rome, Italy

Introduction: Liver regeneration and chronic liver injuryoccurring during HCC development may result in compensatoryproliferation of differentiated hepatocytes. Several lines of evi-dence demonstrated that some pro-inflammatory cytokines (i.e.TNF-a) and gut-derived products, such as lipopolysaccharide (LPS)have been involved in the regulation of hepatocytes proliferationand apoptosis.

Aim: The aim of the present study was to explore the role ofLITAF, an LPS-induced TNF-a transcription factor, in the regulationof hepatocyte proliferation.

Materials and Methods: We established an in vivo model ofhepatocyte proliferation by 70% partial hepatectomy (PHx) in 24male Wistar rats. We used this model to investigate, by real-timePCR, Western Blot and immunohistochemistry analysis, the hepaticexpression of LITAF and of those proteins/genes involved in thecontrol of cell cycle and apoptosis. We further analysed the sameproteins/genes in HepG2 cells stably silenced with shRNA lentivirusfor LITAF.

Results: We found an up-regulation of LITAF mRNAs and pro-tein expression after 1hr and 6 hrs from PHx. This LITAF increasecorrelated with that of it transcriptional regulator, p53 and withan up-regulation of PCNA, a well-known proliferation marker. Noapparent correlation of LITAF with the expression of other cell cyclemarkers, such as cyclin D1 and cyclin B1, was found. These data sug-gest a potential role of LITAF in hepatocyte homeostasis regulation.In fact, the proliferation rate was increased in stable LITAF-silencedHepG2 cells with particular effect at early times of cells cycle, eventhough the absence of LITAF unaffected the expression of cyclinsD1 and B1, cdk4 and cdk1 and TA, DTA and DN isoforms of p53 andapoptosis rate.

Conclusions: In conclusions, this preliminary data indicates apivotal role of LITAF in regulating early times of first cell cycle and

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-47

ARCOPENIA IS ASSOCIATED WITH A REDUCEDURVIVAL IN PATIENTS WITHEPATOCARCINOMA UNDERGOING SORAFENIBREATMENT

. Gigante 1, G. Antonelli 1, P. Begini 1,

. Carbonetti 2, E. Iannicelli 2, P. Marchetti 3,

. Miglioresi 4, A. Pellicelli 4, G. Delle Fave 1,. Marignani 1

Digestive and Liver Diseases unit, St. Andreaospital, Faculty of Medicine and Psychology,niversity Sapienza’, Rome, ItalyRadiology unit, St. Andrea Hospital, Faculty ofedicine and Psychology, University Sapienza’,

ome, ItalyOncology unit, St. Andrea Hospital, Faculty ofedicine and Psychology, University Sapienza’,

ome, ItalyLiver Unit, San Camillo-Forlanini Hospital, Rome,

taly

Background and aim: Sarcopenia has been associated withoor outcomes in patients with cirrhosis and solid tumors, and withigher toxicity during chemotherapy. Hepatocarcinoma (HCC) isiagnosed at advanced stage in up to 40% of all patients. At thistage, the only approved treatment is Sorafenib. We analyzed thenfluence of sarcopenia on survival of HCC patients treated withorafenib.

Methods: We conducted a retrospective study on 41 patientsith advanced HCC treated with Sorafenib. Enrolled patients per-

ormed abdominal computed tomography (CT) 30 days withinreatment start, and data regarding survival, treatment toxicity,nthropometric features, and laboratory findings were collected.transverse CT image from L3 was collected from each scan and

nalyzed with SliceOmatic 5.0. The muscles’ cross-sectional area athis level were selected and normalized for height, obtaining thekeletal-muscle-index (SMI). Presence of sarcopenia was definedy SMI ≤41 cm2/m2 for women, and ≤53cm2/m2 for men withody mass index (BMI) ≥25, and ≤43 cm2/m2 for those with a BMI25.

Results: Patients were mainly males (80,5%). Sarcopenia wasresent in 36,5% of patients (15/41). The two groups, with orithout sarcopenia were compared. Sex distribution and baselines

eatures were similar, no significant differences in albumin, INR,MI, Sodium, creatinine, bilirubin and MELD score were detected.hild-Pugh Score was higher in the sarcopenic group [6 (5-8) vs 55-8); p = 0,034]. Sarcopenic patients had reduced Overall Survival9 (95% CI 44-114) vs 181(95% CI 140-223) weeks (p = 0,0013), asell as smaller survival after treatment: 24 vs 54 weeks(figure 1).

ime on treatment of sarcopenic patients was 12,3 vs 22,9 weeksH.R. 1,83, p = 0,0464), but there were no differences in the cause ofnterruption.

Conclusion: Sarcopenia is present in a third of advanced HCC,
nd is associated with reduced OS and with reduction of time onorafenib treatment.

sease 47S (2015) e43–e66 e65

F-48

ORAL BACTERIAL LOAD IN AUTOIMMUNE LIVERDISEASES: POSSIBLE ROLE OR COINCIDENCE?

G. Orrù 1, S. Onali 2, C. Salustro 2, S. Cappellini 2,E. Cocco 1, S. Fais 1, M. Melis 1, A. Gilardi 1,B. Musu 1, L. Secci 2, G. Serra 2, C. Balestrieri 2,M. Conti 2, M. Casale 2, S. Casu 2, F. Figorilli 2,M.C. Pasetto 2, R. Littera 3, R. Scioscia 2, L. Barca 2,L. Chessa 2

1 DNA Sequencing Service, Department of SurgicalSciences, University of Cagliari, Cagliari, Italy2 Center for the Study of Liver Diseases, Departmentof Medical Sciences “M.Aresu”, University of Cagliari,Cagliari, Italy3 Medical Genetics, Department of Medical Sciences“M.Aresu”, University of Cagliari, Cagliari, Italy

Background and aims: Several factors seem be involved in thepathogenesis of autoimmune liver disease (ALD), such as bacte-ria or bacterial components. Some studies highlighted the role ofan oral bacterium, P. gingivalis, in the pathogenesis of rheumatoidarthritis and in the induction of NASH. We evaluated if there was acorrelation between oral bacteria and some features of ALD.

Methods: We analyzed 50 patients with ALD, 22 with autoim-mune hepatitis (AIH) and 28 with primary biliary cirrhosis (PBC),94% females, median age 61 years, median disease duration 7.5years, 18% with cirrhosis, 66% responders to treatment, 40%with concomitant autoimmune diseases, and 46 health controlsmatched for sex and age. Patients and controls were subjected tocytobrush of the tongue. Exclusion criteria were antibiotic use inthe previous week, mouthwash use on the day of inclusion. Thestudy was conducted with the local ethics committee approvaland informed consent was obtained from all subjects. BacterialDNA (BD) obtained from the tongue biofilm was quantified usinga computer RNA-DNA and expressed as ug/ml of sample.For eachanalysis, three distinct biological replicas were done, and quantita-tive data were expressed as mean ± SD.

Results: BD amount was greater in ALD than in controls(1834.10 vs. 881.75, p < 0.0001). There was no correlation betweenBD amount and age, disease duration, histological findings and bio-chemical parameters, except for alkaline phosphatase (p = 0.05).The presence of cirrhosis, response to therapy and the immunosup-pressive drug use did not affect BD amount. Finally, patients withconcomitant autoimmune diseases had a greater BD amount thanthe other (2053.54 vs. 1687.60) but without significance (p = 0.57).

Conclusion: Our study showed an unspecified role of oral bacte-ria in patients with autoimmune liver diseases. Further analysis

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-49

FFICACY OF SORAFENIB IN PATIENTS WITHNTERMEDIATE-STAGE HEPATOCELLULARARCINOMA: RESULTS FROM THE ITA.LI.CA.ATABASE

. Sacco, V. Mismas, A. Romano, M. Bertini,. Bertoni, G. Federici, G. Parisi, S. Metrangolo,

. Tumino, G. Bresci

Department of Gastroenterology, Pisa Universityospital, Pisa, Italy, for ITA.LI.CA group

Introduction Sorafenib represents the elective treatment foratients affected from advanced-stage (BCLC-C according tohe Barcelona Clinic of Liver Cancer classification) HCC; this

olecule is also recommended in patients with intermediate-tage (BCLC-B) disease who have failed or are not eligible toransareterial chemoembolization (TACE). However, further infor-

ation on the use of sorafenib in patients with BCLC-C HCC isarranted.

Aims This study analyses the efficacy of sorafenib in patientsith BCLC-B HCC, with respect to those with BCLC-C disease, in theation-wide Italian database ITA.LI.CA.

Patients and methods The ITA.LI.CA. database contains data of428 HCC patients treated at 18 Italian Centers. All patients withither BCLC stage B or C and who received sorafenib were consid-red. Overall survival (OS), time to progression (TTP), and diseaseontrol rate (DCR) were evaluated. Safety considerations were alsoerformed.

Results A total of 243 patients were included. Of these, 61ere in BCLC-B stage (29 received ≥1 TACE prior to sorafenib)

nd 182 in BCLC-C. In the overall population, median TTP wasmonths (95%CI 6-9), median OS was 13 months (95%CI 10-

5.5), and DCR was 28.3% (complete response, CR = 1.6%; partialesponse, PR = 8.9%; stable disease, SD = 17.8%). Patients in BCLC-

stage had a median TTP of 10 months (95%CI 6-14), a medianS of 19 months (95%CI 12.5-26.5), and a DCR of 39.1% (CR = 2.4%;R = 9.8%; SD = 26.8%), whereas patients in BCLC-C stage had

median TTP of 7 months (95%CI 6-8), a median OS of 11onths (95%CI 10-15.5), and a DCR of 25.3% (CR = 1.3%; PR = 8.7%;

D = 15.3%). The safety profile of sorafenib was similar in the twoub-populations.

Conclusions These “field-practice” findings suggest thathe administration of sorafenib in BCLC-B patients with HCC

ay be effective and is not associated with any new safetyarning.


sease 47S (2015) e43–e66

F-50

CORRELATION BETWEEN NK CELLS ANDRESPONSE TO TRIPLE HCV-THERAPY WITHBOC/TVR

S. Marenco 1, G. Pieri 1, F. Mazza 1, M. Brunacci 1,S. Labanca 1, L. Bruzzone 1, F. Valentina 1,F. Bozzano 2, F. Marras 3, A. De Maria 3,V. Savarino 1, A. Picciotto 1

1 U.O.s. Diagnosi e Terapia delle Epatopatie, IRCCS,San Martino IST, Genova, Italy2 Istituto G.Gaslini, Genova, Italy3 Centro di Eccellenza per la Ricerca Biomedica,Universita di Genova, Genova, Italy

Background and aims. In our previous study on 28 naïvegenotype-1 patients, we showed that the CD56bright CD16+/− cellpopulations and the expression of both activating and inhibitoryNK-cell receptors were reduced at baseline in patients with sus-tained virological response (SVR), after dual therapy with pegylatedinterferon-alpha and ribavirin (IFN/R). The aim of thos study is toverify the correlation between subset cell populations or activat-ing and inhibitory NK-cell receptors and response to triple therapywith boceprevir (BOC) or telaprevir (TVR).

Methods. We prospectively enrolled 33 patients between Jan2012 and Jan 2013, candidate to triple therapy with BOC/TVR.29/33 patients were male, 31/33 non-responder (NR) to IFN/R,21/33 had cirrhosis. We evaluated NK cells populations, activatingand inhibitory receptors before starting triple therapy (baseline).Statistycal analysis was performed with Mann–Whitney U-test. NKcells subset populations were analyzed by cytofluorometric assay(Becton Dickinson, Mountain View, CA, USA). We used specificmouse anti-human mAb to determine the receptors.

Results. 26 patients were treated with TVR and 7 with BOC.17/33 patients obteined SVR, while 16 did not respond (NR). Wefound no differences in the expression of both activating (NKp30,NKp46, DNAM-1) and inhibitory receptors (CD85j) or the subsetcell populations (CD56dullCD16+, CD56brightCD16+/−) between SVRand NR patients (p > 0,08).

Conclusions. As previously demonstrated, differences in theexpression of activating and inhibitory NK cell receptors contributeto the differential response to dual treatment. This evidence wasnot confirmed in patient treated with triple therapy with BOC/TVR.Therefore, the evaluation of NK cells characteristics at baseline is
useless in predicting virological response to triple therapy withBOC/TVR.

Digestive and Liver Disease 47S (2015) e67


1590-8658/

A.I.S.F. 2015: Abstracts evaluation procedure



journal homepage: www.elsevier .com/locate/dld

Thanks to experts evaluating all the abstracts according to predetermined Clinical and Experimental categories.The experts for the 2015 Annual Meeting are listed below:

Category A. “HEPATITIS C” Clinical/Experimental P.A. Aghemo, Milan - P. Andreone, Bologna - D. Bitetto,

Bergamo - S. Bruno, Milan - V. Di Marco, Palermo - A. Mangia, San Giovanni Rotondo (FG) - G. Missale, Parma - F. Morisco, Naples - P. Pontisso, Padua - G. Taliani, Rome - A.L. Zignego, Florence

Category B. “HEPATITIS B & DELTA” Clinical/Experimental B. Coco, Turin - V. Di Marco, Palermo - C. Ferrari,

Parma - M. Iavarone, Milan - A. Marzano, Turin - F.P. Russo, Padua - G. Taliani, Rome - M. Viganò, Milan

Category C. “ALCOHOLIC LIVER DISEASE” Clinical/Experimental G. Germani, Padua - C. Loguercio, Naples - C. Puoti,

Marino (Rome)

Category D. “NON-ALCOHOLIC FATTY LIVER DISEASE” Clinical/Experimental P. Dongiovanni, Milan - L. Miele, Rome - S. Petta,

Palermo - E. Vanni, Turin

Category E. “AUTOIMMUNE HEPATITIS & BILIARY DISEASE” Clinical/Experimental V. Cardinale, Rome - A. Floreani, Padua - L. Muratori,

Bologna - C. Rigamonti, Novara - F. Rosina, Turin - C. Spirli, New Haven, CT (USA)

Category F. “METABOLIC LIVER DISEASE (Emocromatosi, Wilson, etc.)”

Clinical/Experimental A. Cappon, Padua - S. Fargion, Milan - A. Lleo, Rozzano

(MI) - F. Marra, Florence - G. Svegliati-Baroni, Ancona

Category G. “FIBROSIS” Clinical/Experimental M. Fraquelli, Milan - F. Marra, Florence – M. Parola,

Turin - G. Svegliati-Baroni, Ancona - F. Vizzutti, Florence

Category H. “PORTAL HYPERTENSION” Clinical/Experimental A. Berzigotti, Barcelona (Spain) - F. Campagna,

Padua - P. Caraceni, Bologna - A. Ciaccio, Monza - F. Dell’Era, Milan - V. La Mura, Milan - F. Schepis, Modena

Category I. “CIRRHOSIS & ITS COMPLICATIONS” Clinical/Experimental A. Berzigotti, Barcelona (Spain) - F. Campagna,

Padua - P. Caraceni, Bologna - A. Ciaccio, Monza – F. Dell’Era, Milan - V. La Mura, Milan - F. Schepis, Modena

Category L. “HEPATOCELLULAR CARCINOMA” Clinical/Experimental S. Bhoori, Milan - F. Farinati, Padua - E.G. Giannini,

Genoa - L. Gramantieri, Bologna - E. Gringeri, Padua - M. Iavarone, Milan - G. Missale, Parma- F. Piscaglia, Bologna - E. Villa, Modena

Category M. “HEPATOBILIARY SURGERY & LIVER TRANSPLANTATION”

Clinical/Experimental M. Angelico, Rome - L. Baiocchi, Rome - P. Burra,

Padua - M.F. Donato, Milan - S. Fagiuoli, Bergamo

Category N. “ACUTE LIVER INJURY & HEPATOTOXICITY” Clinical/Experimental M. Fraquelli, Milan - G. Germani, Padua – A. Gasbarrini,

Rome - F. Giannone, Modena

Documents

Digestive and Liver Disease - AISF · Digestive and Liver Disease ... Carlo Catassi, Ancona, Italy Umberto Cillo, Padua, ... Medicine, University of Milano-Bicocca, Milan, Italy