journal of hepatology

JOURNAL OF HEPATOLOGY EDITOR-IN-CHIEF

CO-EDITORS

Didier Samuel

Christopher Day, Peter R. Galle

ASSOCIATE EDITORS

EDITORIAL BOARD

Richard Moreau, France Paolo Angeli, Italy Marina Berenguer, Spain Thomas Baumert, France Pierre-Alain Clavien, Switzerland Ira Jacobson, USA Ansgar Lohse, Germany Derek A. Mann, UK Fabio Marra, Italy Philippe Mathurin, France Vincenzo Mazzaferro, Italy Jorg Petersen, Germany Vlad Ratziu, France Robert S. Schwabe, USA Stefan Zeuzem, Germany Jessica Zucman-Rossi, France

Abraldes, Juan, Canada Ackerman, Zvi, Israel Afford, Simon C, UK Alberti, Alfredo, Italy Alpini, Gianfranco, USA Angulo, Paul, USA Anstee, Quentin M, UK Asselah, Tarik, France Avila, Matıás A, Spain Bedossa, Pierre, France Ben-Ari, Ziv, Israel Berg, Thomas, Germany Bertoletti, Antonio, Singapore Beuers, Ulrich, Germany Blum, Hubert E, Germany Bronowicki, Jean-Pierre, France Bruix, Jordi, Spain Burra, Patrizia, Italy Burroughs, Andrew, UK Buti, Maria, Spain Caldwell, Stephen, USA Calvisi, Diego, Germany Caste´ra, Laurent, France Cortez-Pinto, Helena, Portugal Craxı`, Antonio, Italy Dalekos, Georgious, Greece Dandri, Maura, Germany Dore, Greg, Australia Duclos-Vallee, Jean-Charles, France Duvoux, Christophe, France Factor, Valentina, USA Farrell, Geoff, Australia Feldstein, Ariel, USA Felipo, Vicente, Spain Fickert, Peter, Austria Forner, Alejandro, Spain Forns, Xavier, Spain Garcia-Pagań, Juan Carlos, Spain Garcia-Tsao, Guadalupe, USA Geier, Andreas, Germany Gine`s, Pere, Spain Gores, Gregory, USA Gramantieri, Laura, Italy Graziadei, Ivo, Austria Greten, Tim F, USA Gustot, Thierry, Belgium Hadengue, Antoine, Switzerland Heathcote, Jenny, Canada Heim, Markus, Switzerland Hoshida, Yujin, USA Houghton, Michael, USA Jaeschke, Hartmut, USA Janssen, Harry LA, The Netherlands Kadry, Zakiyah N, USA Kaneko, Shuichi, Japan Kao, Jia-Horng, Taiwan Kaplowitz, Neil, USA Karpen, Saul J, USA Lampertico, Pietro, Italy Lau, George K.K, China Lebrec, Didier, France Lencioni, Riccardo, Italy Locarnini, Stephen, Australia Lohmann, Volker, Germany Lok, Anna, USA Lotersztajn, Sophie, France Lu¨dde, Tom, Germany Majno, Pietro, Switzerland

SPECIAL SECTION EDITORS Focus Scott L. Friedman, USA Daniel Shouval, Israel Hepatology Snapshot Detlef Schuppan, Germany Massimo Levrero, Italy Controversies in Hepatology Graeme Alexander, UK Frontiers in Liver Transplantation Marina Berenguer, Spain International Hepatology Patrick Marcellin, France Josep Llovet, USA Detlef Schuppan, Germany Clinical Application of Basic Science David Adams, UK Scott L. Friedman, USA Web site Elisabetta Bugianesi, Italy Andreas Teufel, Germany

STATISTICAL CONSULTANT Raphael Porcher, France

EDITORS EMERITUS Dame Sheila Sherlock, Founding Editor (1918–2001) Jean-Pierre Benhamou, France (1927–2008) Gustav Paumgartner, Germany Juan Rode´s, Spain Massimo Colombo, Italy

EDITORIAL OFFICE MANAGER Simona Negrini

EDITORIAL COORDINATOR Esther M.N. Dohmann Joe¨l Walicki

MEDICAL ILLUSTRATOR Laurence Zulianello EASL Office Journal of Hepatology Editorial Office 7 rue Daubin 1203 Geneva, Switzerland Tel: (+41) 22 807 03 67 Fax: (+41) 22 510 24 00 e-mail: jhepatology@easloffice.eu

EASL GOVERNING BOARD SECRETARY GENERAL

SCIENTIFIC COMMITTEE

Markus Peck-Radosavljevic, Austria

Matıás A. Avila, Spain Frank Lammert, Germany Helen Louise Reeves, UK Alessio Aghemo, Italy Cecilia Rodrigues, Portugal

VICE SECRETARY

Laurent Castera, France TREASURER

Mauro Bernardi, Italy

EDUCATIONAL COUNCILLORS

Jean-François Dufour, Switzerland Cihan Yurdaydin, Turkey EU POLICY COUNCILLOR

Patrizia Burra, Italy

Mann, Jelena, UK Manns, Michael P, Germany Marchesini, Giulio, Italy Marschall, Hanns-Ulrich, Sweden Mato, Jose, Spain Merkel, Carlo, Italy Mieli-Vergani, Giorgina, UK Monga, Satdarshan S, USA Moshage, Han, The Netherlands Mu¨llhaupt, Beat, Switzerland Mutimer, David, UK Negro, Francesco, Switzerland Neuberger, James, UK Oakley, Fiona, UK Oben, Jude, USA O’Grady, John, UK Oude Elferink, Ronald, The Netherlands Pageaux, Georges-Philippe, France Pares, Albert, USA Patel, Keyur, USA Pawlotsky, Jean-Michel, France Perrillo, Robert, USA Pianko, Stephen, Australia Pietroangelo, Antonello, Italy Pietschmann, Thomas, Germany Porte, Robert J, The Netherlands Prati, Daniele, Italy Prieto, Jesu´s, Spain Puoti, Massimo, Italy Raimondo, Giovanni, Italy Rehermann, Barbara, USA Renner, Eberhard, Canada Rizzetto, Mario, Italy Rogiers, Xavier, Germany Rosenbaum, Jean, France Roskams, Tania, Belgium Rosmorduc, Olivier, France Runyon, Bruce A, USA Safadi, Rifaat, Israel Salerno, Francesco, Italy Saliba, Faouzi, France Sarrazin, Christoph, Germany Sauerbruch, Tilman, Germany Schwartz, Myron, USA Spengler, Ulrich, Germany Stickel, Felix, Switzerland Stieger, Bruno, Switzerland Strassburg, Christian P, Germany Talal, Andrew, USA Taylor Robinson, Simon, UK Thimme, Robert, Germany Thomas, David, USA Tilg, Herbert, Austria Tillmann, Hans, USA Tong, Shuping, USA Trautwein, Christian, Germany Trinchet, Jean-Claude, France Trotter, James F, USA Valla, Dominique, France Vergani, Diego, UK Villanueva, Augusto, Spain Wedemeyer, Heiner, Germany Weiland, Ola, Sweden Wiest, Reiner, Germany Yeh, Chau-Ting, Taiwan Zarsky, Jean-Pierre, France Zuckerman, Eli, Israel

Applications for membership of EASL, together with a letter of reference, should be sent to EASL office, 7 rue Daubin, CH 1203, Geneva, Switzerland. Tel.: +41 22 807 03 60; fax: +41 22 328 07 24; e-mail: easloffice@easloffice.eu © 2014 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved. Important notice to authors: The work shall not be published elsewhere in any language without the written consent of the Journal of Hepatology. The articles published in this journal are protected by copyright, which covers translation rights and the exclusive right to reproduce and distribute all of the articles printed in the journal. No material published in the journal may be stored on microfilm or videocassettes or in electronic databases and the like or reproduced photographically without the prior written permission of Journal of Hepatology. Publication information: Journal of Hepatology (ISSN 0168-8278). For 2014, volumes 60 and 61 are scheduled for publication. Subscription prices are available upon request from the Publisher or from the Elsevier Customer Service Department nearest you or from this journal’s website (http://www.elsevier.com/locate/jhep). Further information is available on this journal and other Elsevier products through Elsevier’s website: (http://www.elsevier.com). Subscriptions are accepted on a prepaid basis only and are entered on a calendar year basis. Issues are sent by standard mail (surface within Europe, air delivery outside Europe). Priority rates are available upon request. Claims for missing issues should be made within six months of the date of dispatch. USA mailing notice: Journal of Hepatology (ISSN 0168-8278) is published monthly by Elsevier B.V. (Radarweg 29, 1043 NX Amsterdam, The Netherlands). Periodicals postage paid at Jamaica, NY 11431 and additional mailing offices (not valid for journal supplements). 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Acknowledgements The European Association for the Study of the Liver thanks all the abstract reviewers for their time and effort and appreciates their contribution towards the success of the International Liver Congress™ 2014: David Adams, UK Kosh Agarwal, UK Alessio Aghemo, Italy Guruprasad Padur Aithal, UK Ulus Akarca , Turkey Matthew Albert, France Luis Alvarez, Spain Raul Jesus ´ Andrade , Spain Pietro Andreone, Italy Tarik Asselah, France Birke Bartosch, France Jacques Belghiti, France Thomas Berg, Germany Gabriela Berlakovich, Austria Annalisa Berzigotti, Spain Ulrich Beuers, The Netherlands Massimo Bologniesi, Italy Ashley Brown, UK Elisabetta Bugianesi, Italy Christophe Bureau, France Maria Buti, Spain Ali Canbay, Germany Evangelos Cholongitas, Greece Thierry Claudel, Austria Gonzalo Crespo, Spain George Dalekos, Greece Olav Dalgard, Norway Harry Dalton, UK Maura Dandri, Germany Christian Datz, Austria Andrea De Gottardi, Switzerland Samuele De Minicis, Italy Anil Dhawan, UK Helmut Diepolder, Germany Steven Dooley, Germany Robert Eferl, Austria Isabel Fabregat, Spain

Fabio Farinati, Italy Jordan Feld, Canada Arnulf Ferlitsch, Austria Robert Flisiak, Poland Alejandro Forner, Spain Graham Foster, UK Claire Francoz, France Mirella Fraquelli, Italy Mireen Friedrich-Rust, Germany Juan Carlos Garcia Pagan, Spain Jacob George, Australia Giacomo Germani, Italy David Goldberg, UK Ivo Graziadei, Austria Jochen Hampe, Germany Markus Heim, Switzerland Claus Hellerbrand, Germany Gideon Hirschfield, Canada Harald Hofer, Austria Massimo Iavarone, Italy Jerzy Jaroszevic, Poland Mickael Lesurtel, Switzerland Christian Liedtke, Germany Volker Lohmann, Germany Sophie Lotersztajn, France Beat Mullhaupt, ¨ Switzerland Yun Ma, UK Mariana Machado, Portugal Pietro Majno, Switzerland Vincent Mallet, France Alessandra Mangia, Italy Derek Mann, UK Patrick Marcellin, France Maria Martinez-Chantar , Spain Mari Montserrat, Spain Jacob Natterman, Germany Frederic Nevens, Belgium

Johan Neyts, Belgium Valerie ´ Paradis, France Rosa M. Pascal, Italy Thomas Pietschmann, Germany Anne-Christine Piguet, Switzerland Matthias Pinter, Austria Massimo Pinzani, UK Fabio Piscaglia, Italy Stanislas Pol, France Teresa Pollicino, Italy Daniele Prati, Italy Barbara Rehermann, USA Thomas Reiberger, Austria Natalyia Rohr-Udilova, Austria Floriano Rosina, Italy Francoise ¸ Roudot, France Ian Rowe, UK Jose Maria Sanchez-Tapias, Spain Bruno Sangro, Spain Pablo Sarobe, Spain Thomas Scherzer, Austria Nasser Semmo, Switzerland Wolfgang Sieghart, Austria Rudolf Stauber, Austria Felix Stickel, Switzerland Konrad Streetz, Germany Camille Sureau, France Mark Thursz, UK Sarah Toffani, Germany Jonel Trebicka, Germany Jorg ¨ Trojan, Germany Florian van Bommel, Germany Valerie ´ Vilgrain, France Augusto Villanueva, Spain Wolfgang Vogel, Austria Christoph Welsch, Germany Jessica Zucman-Rossi, France

JOURNAL OF HEPATOLOGY VOLUME 60, SUPPLEMENT 1, PAGES S1–S598

Abstracts of The International Liver Congress™ 2014 – 49th annual meeting of the European Association for the Study of the Liver April 9–13, 2014, London, United Kingdom

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Changing Dynamics in the Science and Clinical Management of Primary Biliary Cirrhosis: What They Mean for Us, What They Mean for Our Patients

A Satellite Symposium of the International Liver CongressTM 2014

Saturday, 12 April, 18:00 Capital Suite 2-3-4, ExCel London Refreshments will be served before the symposium

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CHAIR

Introduction and Stage Setting

David E.J. Jones, MD, PhD — David E.J. Jones, MD, PhD

Recent Advances in PBC: Disease Pathways and Potential Targets of Therapy — Ulrich H.W. Beuers, MD Challenges and New Opportunities in the Clinical Management of PBC — David E.J. Jones, MD, PhD Question and Answer Session

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FACULTY Ulrich H.W. Beuers, MD Professor Department of Gastroenterology & Hepatology Tytgat Institute for Liver and Intestinal Research University of Amsterdam Amsterdam, The Netherlands

— Faculty Panel

Concluding Remarks — David E.J. Jones, MD, PhD

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JOURNAL OF HEPATOLOGY VOLUME 60, SUPPLEMENT 1, PAGES S1–S598 CONTENTS Acknowledgements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

iv

Thursday, 10 April 2014 OPENING & GENERAL SESSION 1: VIRAL HEPATITIS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S1

Parallel Session

S4

HEPATITIS C TREATMENT . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

Parallel Session

PORTAL HYPERTENSION: CIRRHOSIS BLEEDING, HEPATORENAL ASCITES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S7

Parallel Session

FATTY LIVER DISEASE: CLINICAL AND EXPERIMENTAL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S10

Parallel Session

IASL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S13

Parallel Session

LIVER TRANSPLANTATION: CLINICAL AND EXPERIMENTAL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S17

Parallel Session

HEPATITIS C: EXPERIMENTAL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S20

GENERAL SESSION 2 & AWARDS 1: GENERAL HEPATOLOGY . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S23

Parallel Session

HEPATITIS C: NEW ANTIVIRAL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S25

Parallel Session

COMPLICATIONS OF CIRRHOSIS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S29

Parallel Session

NON INVASIVE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S32

Parallel Session

PUBLIC HEALTH . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S35

Parallel Session

LIVER TUMOURS: EXPERIMENTAL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S38

Parallel Session

HEPATITIS B & D: EXPERIMENTAL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S41

Friday, 11 April 2014

Saturday, 12 April 2014 GENERAL SESSION 3 & AWARDS 2: LIVER TUMORS AND TRANSPLANTATION . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S45

Parallel Session

HEPATITIS B & E: CLINICAL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S48

Parallel Session

LIVER TUMOURS MANAGEMENT . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S51

Parallel Session

CHOLESTATIC AND GENETIC LIVER DISEASES. PEDIATRICS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S54

Parallel Session

ALCOHOL AND DRUG INDUCED LIVER DISEASES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S59

Parallel Session

MOLECULAR AND CELLULAR BIOLOGY: HSC FIBROSIS, CIRRHOSIS, PATHOPHYSIOLOGY . . . . . . . . . . . . . . . . . . . . . . . . . .

S61

Parallel Session

LIVER IMMUNOLOGY . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S64

Poster Session – Thursday, 10 April 2014 1A. LIVER TRANSPLANTATION/SURGERY: EXPERIMENTAL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S67

2A. CIRRHOSIS AND ITS COMPLICATIONS: PATHOPHYSIOLOGY . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S71

3A. LIVER TUMORS: EXPERIMENTAL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S81

4A. MOLECULAR AND CELLULAR BIOLOGY: CELL CYCLE CONTROL/APOPTOSIS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S99

5. LIVER IMMUNOLOGY INCLUDING VIRAL HEPATITIS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S106

7A. VIRAL HEPATITIS: HEPATITIS B & D – EXPERIMENTAL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S122

8A. VIRAL HEPATITIS: HEPATITIS C – EXPERIMENTAL (VIROLOGY)

S131

..............................................................................

10A. FATTY LIVER DISEASE: EXPERIMENTAL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S143

11. ALCOHOLIC LIVER DISEASE AND DRUG INDUCED LIVER DISEASE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S164

12. AUTOIMMUNE AND CHRONIC CHOLESTATIC LIVER DISEASE . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S182

15. IASL

S202

......................................................................................................................................

Poster Session – Friday, 11 April 2014 2B. CIRRHOSIS AND ITS COMPLICATIONS: CLINICAL ASPECTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S215

3B. LIVER TUMORS: CLINICAL (EPIDEMIOLOGY, DIAGNOSIS) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S250

4B. MOLECULAR AND CELLULAR BIOLOGY: HSCS AND FIBROSIS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S266

7B. VIRAL HEPATITIS: HEPATITIS A, B, D & E CLINICAL (EXCEPT THERAPY) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S280

8B. VIRAL HEPATITIS: HEPATITIS C – CLINICAL (EXCEPT THERAPY) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S305

10B. FATTY LIVER DISEASE: CLINICAL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S334

Poster Session – Saturday, 12 April 2014 1B. LIVER TRANSPLANTATION/SURGERY: CLINICAL . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S361

2C. CIRRHOSIS AND ITS COMPLICATIONS: BLEEDING, HEPATORINAL SYNDROME AND ASCITIS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S384

3C. LIVER TUMORS: MANAGEMENT . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S394

6. NON-INVASIVE MARKERS OF LIVER FIBROSIS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S409

7C. VIRAL HEPATITIS: HEPATITIS B & D – CLINICAL (THERAPY, NEW COMPOUNDS, RESISTANCE) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S421

8C. VIRAL HEPATITIS: HEPATITIS C – CLINICAL (THERAPY) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S443

8D. VIRAL HEPATITIS: HEPATITIS C – CLINICAL (NEW COMPOUNDS, RESISTANCE) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S496

13. GENETIC AND PEDIATRIC LIVER DISEASES . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S504

14. EU PUBLIC HEALTH . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S513

Thursday, 10 April – Saturday, 12 April, 2014 Parallel Session

LATE-BREAKER ORAL PRESENTATIONS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S523

LATE-BREAKER POSTER ABSTRACTS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S527

Author index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Disclosures: no commercial relationships . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . Disclosures: commercial relationships . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

S539 S591 S597

Registration of Clinical Trials The Journal of Hepatology endorses the policy of the WHO and the International Committee of Medical Journal Editors (ICMJE) on the registration of clinical trials. Therefore, any trial that starts recruiting on or after July 1, 2005 should be registered in a publicly owned, publicly accessible registry and should satisfy a minimal standard dataset. Trials that started recruiting before that date will be considered for publication if registered before September 13, 2005. More detailed information regarding clinical trials and registration can be found in New Engl J Med 2004; 351:1250–1251 and New Engl J Med 2005; 352:2437–2438.

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ORAL PRESENTATIONS

Thursday, 10 April 2014

OPENING & GENERAL SESSION 1: VIRAL HEPATITIS

O1 SAPPHIRE II: PHASE 3 PLACEBO-CONTROLLED STUDY OF INTERFERON-FREE, 12-WEEK REGIMEN OF ABT-450/R/ABT-267, ABT-333, AND RIBAVIRIN IN TREATMENT-EXPERIENCED ADULTS WITH HEPATITIS C VIRUS GENOTYPE 1 S. Zeuzem1 , I. Jacobson2 , T. Baykal3 , R.T. Marinho4 , F. Poordad5 , M. Bourliere6 , M. Sulkowski7 , H. Wedemeyer8 , E. Tam9 , P. Desmond10 , D. Jensen11 , A.M. Di Bisceglie12 , P. Varunok13 , T. Hassanein14 , J. Xiong3 , B. DaSilva-Tillmann3 , L. Larsen3 , T. Podsadecki3 . 1 J.W. Goethe University, Frankfurt, Germany; 2 Weill Cornell Medical College, New York, NY, 3 AbbVie Inc., North Chicago, IL, United States; 4 Centro Hospitalar de Lisboa Norte, Lisbon, Portugal; 5 The Texas Liver Institute, University of Texas Health Science Center, opital Saint Joseph, Marseille, San Antonio, TX, United States; 6 Hˆ France; 7 Johns Hopkins University, Baltimore, MD, United States; 8 Medizinische Hochschule Hannover, Hannover, Germany; 9 LAIR Centre, Vancouver, BC, Canada; 10 St Vincent’s Hospital (Melbourne), Fitzroy, VIC, Australia; 11 Center for Liver Diseases, University of Chicago Medical Center Chicago, Chicago, IL, 12 Saint Louis University, St. Louis, MO, 13 Premier Medical Group of the Hudson Valley, PC, Poughkeepsie, NY, 14 Southern California Liver Centers and Southern California Research Center, Coronado, CA, United States E-mail: [email protected] Background and Aims: Efficacy in retreatment of HCV-infected patients is associated with treatment history, with the lowest responses occurring among prior peginterferon/ribavirin nullresponders. ABT-450 is an HCV NS3/4A protease inhibitor (dosed with ritonavir 100 mg, ABT-450/r) identified by AbbVie and Enanta. ABT-267 is an NS5A inhibitor; ABT-333 is an NS5B RNA polymerase inhibitor. The safety and efficacy of ABT-450/r/ABT-267 + ABT-333+RBV (3D+RBV) was evaluated among non-cirrhotic peginterferon/ribavirin-experienced, HCV genotype (GT)1-infected patients in an interferon-free phase III trial. Methods: In this double-blind, placebo-controlled trial, prior peginterferon/ribavirin relapsers, partial-responders, or null-responder patients were randomized (3:1) to receive ABT-450/r/ABT-267 (150 mg/100 mg/25 mgQD) + ABT-333 (250 mgBID) + weight-based RBV (Arm A) or matching placebos (Arm B) for 12 weeks. Results: 297 patients received 3D+RBV; 97 received matching placebos. The Arm A (3D+RBV active regimen) SVR12 rate was 96.3% (286/297). 2.4% of patients had virologic failure. SVR12 rates in prior peginterferon/ribavirin relapsers, partial-responders, and null-responders were 95.3%, 100%, and 95.2%, respectively. SVR12 rates were comparable among genotype 1a and 1b patients (96.0% and 96.7%, respectively). The most common adverse events (AEs) in Arm A (3D+RBV active regimen) and Arm B (placebo) were headache (36.4% and 35.1%, respectively) and fatigue (33.3% and 22.7%, respectively); the frequency of these events did not differ significantly between arms (P > 0.05). No moderate/severe AEs occurred significantly more

frequently in Arm A vs. B (P > 0.05). Rates of discontinuation due to AEs were 1.0% and 0% in Arm A (3D+RBV active regimen) and Arm B (placebo), respectively. Conclusions: A multi-targeted antiviral approach combining ritonavir-boosted ABT-450, ABT-267, and ABT-333 with ribavirin achieves high SVR12 rates with low rates of treatment discontinuation in treatment-experienced non-cirrhotic HCV genotype 1-infected patients, including prior null-responders. Table: Baseline characteristics and efficacy

Male, n (%) White race, n (%) HCV GT1a, n (%)* IL28B CC, n (%)

Fibrosis score ≥F2† , n (%) Baseline HCV RNA level, log10 IU/mL, mean (SD) Prior relapser, n (%) Prior partial-responder, n (%) Prior null-responder, n (%) SVR12 rate, n/N (%)‡ All patients HCV GT1a-infected patients HCV GT1b-infected patients On treatment virologic failure, n/N (%) Relapse post treatment, n/N (%) Premature study drug discontinuation, n/N (%)

Arm A (3D+RBV), N = 297

Arm B (Placebo), N = 97

167 (56.2) 269 (90.6) 173 (58.2) 34 (11.4)

60 (61.9) 86 (88.7) 57 (58.8) 7 (7.2)

95 (32.0) 6.55 (0.54) 86 (29.0) 65 (21.9) 146 (49.2)

32 (33.0) 6.52 (0.48) 29 (29.9) 21 (21.6) 47 (48.5) −

286/297 (96.3)§ 166/173 (96.0) 119/123 (96.7) 0 7/293 (2.4) 4/297 (1.3)

− − −

*Subgenotype could not be determined for 1 patient. † Fibrosis stage was assessed by liver biopsy scores, FibroScan scores, or FibroTest scores mapped to Metavir equivalent scoring. ‡ Patients missing data in SVR12 window count as failures. § Superior to the historical SVR rate for telaprevir and peginterferon/ribavirin.

O2 OBETICHOLIC ACID, A FARNESOID-X RECEPTOR AGONIST, REDUCES BACTERIAL TRANSLOCATION AND RESTORES INTESTINAL PERMEABILITY IN A RAT MODEL OF CHOLESTATIC LIVER DISEASE L. Verbeke1 , R. Farre2 , K. Covens3 , B. Verbinnen4 , T. Vanuytsel5 , I. Vander Elst1 , P. Windmolders1 , J. Verhaegen6 , M. Komuta7 , T. Roskams7 , J. Trebicka8 , F. Nevens1 , W. Laleman1 . 1 Department of Liver and Biliopancreatic Disorders, KU Leuven, 2 Translational Research Center for Gastrointestinal Disorders, 3 Molecular and Vascular Biology, 4 Experimental Immunology, 5 Translational Research Center for Gastrointestinal Disorders, 6 Microbiology, 7 Pathology, KU Leuven, Leuven, Belgium; 8 Department of Internal Medicine I, University of Bonn, Bonn, Germany E-mail: [email protected] Background and Aims: Bacterial translocation (BT) drives the pathogenesis and complications of cirrhosis. The farnesoid-X receptor (FXR) is considered a key transcription regulator involved in intestinal barrier function, bile and liver metabolism. We studied potential intestinal FXR-dysfunction in a cholestatic animal model and evaluated the impact of obeticholic acid (INT-747), an FXRagonist, on BT, gut permeability, inflammation and liver damage. Methods: Rats were gavaged with INT-747 5 mg/kg or vehicle every 2 days during 10-day bile-duct ligation (BDL; n = 14+20) and assessed for changes in BT (by culture of ascites and mesenteric lymph nodes (MLN)), permeability (Ussing-chambers), tightjunction protein expression, intestinal immune cell recruitment and cytokine-expression (by FACS + RT-PCR in ileum, MLN and spleen) and liver injury. Healthy rats served as control (n = 17). Auxiliary

Journal of Hepatology 2014 vol. 60 | S1–S22 © 2014 All rights reserved.

ORAL PRESENTATIONS in-vitro BT-mimicking experiments, using the CaCo2 (human enterocyte-like)-cell line were performed in TranswellsTM . Results: Vehicle-treated BDL-rats showed reduced ileal FXR pathway expression (P = 0.02 vs. healthy control) which was associated with increased BT, ileal permeability (through increased claudin-2-expession), recruitment of macrophages, natural killer and dendritic cells and interferon-g (IFN-g)expression (P < 0.05 vs. control). Following INT-747-treatment, immune cell recruitment and IFN-g expression were markedly reduced which was associated with normalized permeability (by up-regulated claudin-1, occludin and zonula-occludens-protein-1) (P < 0.05 vs. BDL+vehicle). Following INT-747-treatment plasma bilirubin decreased from 12.4±0.4 to 8.4±0.7 mg/dl in BDL-rats (P < 0.01). In vitro, IFN-g induced increased permeability and E. coli translocation which remained unaffected with INT-747preincubation. Conclusions: In experimental cholestasis, FXR-agonism restores ileal permeability and decreases BT by attenuating intestinal inflammation, demonstrating a crucial protective role for FXR in the gut-liver axis.

the add-on versus 10 (11%) in the monotherapy-arm achieved HBeAg-seroconversion with HBV DNA 9.3 IU/mL but 0.28 for histology P < 0.01 and >0.49 for blood-tests P < 0.0001), which underlined the risk of low specificity. In multivariate models [multivariate regressioncoefficient (MRC 95% CI; P-value)], CK18-cleaved/CK18-full had similar profiles, with strong associations with activity-SAF [0.09 (0.02–0.17) P = 0.01 / 0.10 (0.03–0.17) P = 0.009], and ActiTest [0.69 (0.51–0.87) / 0.72 (0.55–0.89); both P < 0.0001] adjusted on steatosis/fibrosis. Associations CK18-cleaved/CK18-full with steatosis-SAF/SteatoTest were strongly reduced when adjusted on activity-SAF/fibrosis-SAF but remained significant for steatosis-SAF/ SteatoTest [0.12 (0.03–0.20) P = 0.007 / 0.28 (0.15–0.41) P < 0.0001]. Associations CK18-cleaved/CK18-full with fibrosis-SAF/FibroTest were no more significant when adjusted on activity-SAF/ steatosis-SAF. miR-122 profile was different with lower associations with SAF/blood-tests (R2 = 0.28 vs 0.41 for CK18-cleaved and 0.52 for CK18-full). Association with activity-SAF persisted [MRC = 0.14 (7–20) P = 0.0001] after fibrosis-SAF or steatosis-SAF adjustment (0.11 (0.04–0.18) P = 0.002) but not with both. Same conclusions were obtained using biopsy or FibroTest-ActiTest-SteatoTest. Conclusions: DILI-candidates such as cleaved/full-lengthcytokeratin-18 and miR-122 are highly correlated with activity, but also associated with steatosis and fibrosis. Specificity of such candidates must be assessed and compared in appropriate controls before qualification. P328 ROLE OF MYCOPHENOLATE MOFETIL (MMF) IN STEROID NON RESPONSIVE SEVERE ACUTE ALCOHOLIC HEPATITIS: A RANDOMIZED OPEN LABEL PLACEBO CONTROL PROSPECTIVE CLINICAL PILOT TRIAL – MASH TRIAL P.P. Basu1,2 , M. Aloysius2 , N.J. Shah3 , R.S. Brown Jr1 . 1 College of Physicians and Surgeons, Columbia University, 2 King’s County Hospital Medical Center, New York, 3 James J. Peters VA Medical Center, Icahn School of Medicine at Mount Sinai, Bronx, NY, United States E-mail: [email protected] Background and Aims: Severe alcoholic hepatitis (SAH) has a mortality of 47% with steroid non-responsiveness. Immunosuppressives remains SOC. Mycophenolate mofetil (MMF) is a immunosuppressive reduces B and T lymphocytic recruitment and immune injury.

Journal of Hepatology 2014 vol. 60 | S67–S214

S175

POSTERS Methods: Thirty patients aged-25–60 diagnosed with steroid unresponsive SAH, with severity score criteria (DF > 32%, ABCI > 12, Lille > 0 .45, MELD > 26. Exclusion; hepatitis A, BC, Gi bleed, hepatic Encephalopathy sepsis, Group A MMF 500 mg bid with Ptx 400 mg and Placebo orally for 30 days. Group B Prednisolone 30 mg plus Placebo (Vit D) and Ptx 400 mg orally 30 days. Group C Prednisolone plus NMF and PTX orally for 30 days. Results: See the table. Group A

T. bilirubin INR Albumin Creatinine Platelet AST/ALT TNF IL-6 IL-8

Group B

Group C

Day 0

Day 90

Day 0

Day 90

Day 0

Day 90

14 2.1 2.3 1.8 138 321/187 9.7 7.9 5.3

1.3 0.9 3.3 0.7 303 31/23 0.4 0.2 0.03

13 2.1 2.4 1.5 121 341/157 8.9 6.7 4.9

8 1 3 0.8 251 39/25 1.1 1 0.6

15 2.3 1.9 2.4 98 421/299 12.2 9.8 11.4

2.1 0.8 3.1 1 187 34/36 0.3 0.1 0.8

Conclusions: This study postulates MMF is well tolerated, safe, and effective. MMF independently and precipitously reduced the necroinflammatory score compared with the SOC. A larger trial needs to validate the singular role of MMF in AAH. P329 BASELINE LIVER BIOPSY CAN PREDICT RESPONSE TO STEROIDS IN PATIENTS WITH SEVERE ALCOHOLIC HEPATITIS S.M. Shasthry1 , A. Rastogi2 , A.S. Bhadoria3 , C. Vashishtha1 , M. Kumar1 , S.K. Sarin1 . 1 Hepatology, 2 Hepatopathology, 3 Epidemiology, Institute of Liver & Biliary Sciences, New Delhi, India E-mail: [email protected] Background and Aims: Patients of severe alcoholic-hepatitis (ASH) have high short term mortality and variable response to steroids (30–60%); which is difficult to predict based on clinical or biochemical profile. We evaluated the role of baseline liver biopsy in predicting response to steroids. Methods: Thirty patients [M: F-30:0; age37±14yrs] with biopsy proven ASH and discriminant factor >32 were enrolled. All patients received 40 mg/d of prednisolone, and at day7 were categorized into responders and non-responders by Lille score < or >0.45 respectively. Histological severity of ASH was assessed by a single hepatopathologist (blinded from the clinical response to steroids). Ballooned hepatocytes and lobular inflammation were scored on a 0–5 scale and graded between 0–2. Canalicular cholestasis, perivenular steatosis, Mallory–Denk bodies (MDBs), perivenular fibrosis, ductular cholestasis and cholangiolitis were semi quantitatively graded (Table 1). Table 1. Histological parameters of ASH (semiquantitative) Parameter

Steroid responder, Median (IQR)

Steroid non-responder, Median (IQR)

p value

Ballooning degeneration Lobular inflammation Perivenular steatosis Mallory–Denk bodies Perivenular/pericellular fibrosis Cholangiolitis Canalicular/ductular bile plugs Severity score Grade of alcoholic hepatitis

3 1 1 2 2 0 1 3 1

4 2 1 3 3 0 1 4 2

0.04 0.41 0.77 0.02 0.10 0.34 0.70 0.04 0.03

(2–4) (1–2) (0–2) (1–2) (2–2) (0–1) (1–2) (2–4) (1–2)

(4–5) (1–2) (0–2) (2–3) (2–3) (0–0) (0–2) (4–5) (2–2)

Results: Of 30 patients, 23 (76.7%) were responders. No significant difference in age, DF, MELD, MELDNa, CTP or HVPG was found between responders and non-responders. Non-responders had histologically more severe ASH. Extent of ballooning (grade4 ballooning in 85.7% vs 34.8%; p = 0.042) and MDBs (grade3 MDBs S176

in 71.4% vs 17.4%; p = 0.016) were higher on baseline liver biopsy in non-responders. Other histological parameters showed no significant difference (Table 1). Conclusions: Non-responders to steroids had a greater histological severity (higher ballooning and MDBs) of alcoholic hepatitis. Baseline clinical and biochemical parameters fail to predict nonresponse to steroids. Liver biopsy and histological severity of ASH is important for predicting response to steroids in severe ASH. P330 ACTIVATION OF INTERFERON AND NF-úB PATHWAYS IN EARLY HUMAN ALCOHOLIC LIVER DISEASE DEVELOPMENT 1,2 , N. Lanthier1,2 , C. De Saeger1 , C. Sempoux3 , I.A. Leclercq1 , P. Starkel ¨ Y. Horsmans1,2 . 1 Laboratory of Hepato-Gastroenterology, Université Catholique de Louvain, Institut de Recherche Expérimentale et Clinique, 2 Service d’Hépato-Gastroentérologie, 3 Service d’Anatomie Pathologique, Université Catholique de Louvain, Cliniques Universitaires Saint-Luc, Brussels, Belgium E-mail: [email protected] Background and Aims: Animal data suggest a role for TLR4 with subsequent activation of NF-úB in alcohol liver injury. Here, we assessed cytokines, TLR and TLR-dependent pathways expressions in human ALD. Methods: Liver biopsies from 41 actively drinking ALD patients were compared with normal liver tissue (n = 6) and with liver biopsies from ALD patients after 2 weeks of abstinence (n = 27). Quantitative PCR and Western blotting were used for mRNA and protein analysis, respectively, immunohistochemistry for cellular localization of inflammatory processes. Our local ethics committee has approved the study. Results: Compared with normal livers, already early ALD (fibrosis 80% confluence on collagen-I-coated plates and treated (in triplicates) 24 h with prototypical inducers rifampicin (CYP3A4) and omeprazole (CYP1A2), [n = 3]. 50mM testosterone or phenacetin were added and supernatant and cell samples taken after 2 hours of incubation at 37°C. CYP1A2/3A4 activity [P450-Glo™-Luminometry; Promega] was determined (Figure 1). Relative turnover of testosterone [HPLC] and phenacetin [LCMS/MS] metabolites was also measured. Cell phenotype was assessed by light-microscopy, histology (PAS-Glycogen), CYP3A4, F-actin/phalloidin, and JC-1 fluorescent-staining. Results: Figure 1 shows HepaRG CYP1A2/3A4 activity was 40–80 fold >> HepG2/C3A cells [P < 0.001]; Drug profiling revealed HepaRGs had both enhanced production of major metabolites of phenacetin and testosterone and more intact drug metabolism compared with HepG2/C3A. In contrast with HepG2/C3A, HepaRGs displayed a more intact hepatic phenotype, including: Strong positive glycogen, CYP3A4 staining, high JC-1-positive intrinsic metabolic activity (DYm) and organotypic gross morphology.

Figure 1. Relative activity of CYP1A2 and CYP3A4 enzymes in HepaRG and C3A cells under induction by omeprazole (OMP, CY1A2) or rifampicin (RIF, CYP3A4) controlled against additional enzyme-specific inhibition by fluvoxamin (FLU, CYP1A2) or ketoconazole (KETO, CYP3A4). Activity was measured by luminometry and was normalized internally against appropriate controls. Data are displayed as mean ± standard deviation of % relative activity, normalized to the activity of HepaRG cells under induction (= 100%) and derived from n = 3 biological and n = 8 technical replicates each.

Conclusions: HepaRG cells may represent a more physiologicallyrelevant pre-clinical platform for CYP450 activation/inhibition, safety pharmacology, as well as drug-drug interaction studies. P332 DEVELOPMENT OF HUMAN HEPATIC HepaRG CELL-BASED LiverBioChip PROTOTYPES USING ELECTRON BEAM LITHOGRAPHY DIRECTED-NANOPATTERNING FOR HTSCOMPATIBLE PRE-CLINICAL DRUG EVALUATION APPLICATIONS L.J. Nelson1 , C. LeBled1 , P. Treskes1 , M. Thompson2 , M. Navarro1 , J.N. Plevris1 , N. Gadegaard2 . 1 Hepatology, University of Edinburgh, Edinburgh, 2 James Watt Nanofabrication Centre, University of Glasgow, Glasgow, United Kingdom E-mail: [email protected] Background and Aims: Cells are known to respond to nanoscale features thus our aim was to assess phenotypic morphology of HepaRGs cultured on 2D-nanopatterned polymer slides (NPS) for the optimisation of prototype LiverBioChips for pre-clinical drugscreening. Methods: NPS were manufactured using electron-beam lithography and injection moulding. Slides comprised 11 discrete areas representing varying degrees of nanoscale symmetry and disorder (including 1x planar surface as control). Human hepatic HepaRG and HepG2/C3A control cells were seeded in culture

media, on NPS substrates at high (standard; 2.4×105 /cm2 or 5.6×104 /cm2 respectively) intermediate (75% of high) and low (50%) seeding densities. Cells were cultured for up to 6 days; and morphological phenotype analyzed on days 3 and 6, using light microscopy; LIVE/DEAD viability-assay and ‘Profiler Panel’ immunostaining comprising CYP3A4/phalloidinTRITC/DAPI fluorochromes. Captured images were analysed for cell morphometrics using CellProfiler software. Results: HepaRG cells on two different nanotopographies displayed clear differences in cell attachment/-spreading and enhanced maturation (Figure 1). We observed pattern-dependent early appearance of bile-canaliculae. Moreover, HepaRG differentiated phenotype was observed earlier (day 3) and at lower (intermediate) seeding densities compared with high-density HepaRGs cultured on standard collagen-I substrate (day 6). In contrast HepG2/C3As at all seeding densities did not respond to nanopatterning, maintaining a typical phenotype. Preliminary image analysis of cell morphometrics, could delineate cell numbers/neighbours, as well as relative area/intensity of CYP3A4 staining in HepaRG cultures.

Figure 1. HepaRG cells seeded on two different nanotopographies (DSQ or HEX patterns as indicated) displaying clear differences in proliferation rate and enhanced lineage determination suggesting relative differentiation/ cells numbers are influenced by nanopatterned substrates. Shown are side-by-side displays of HepaRG cells after indicated nanopattern-culture and dual staining via DAPI and phalloidin at 20× magnification and after triple staining including CYP3A4 at 10× magnification. Note punctuate phalloidin-staining of F-actin bands indicative of bile-canaliculae.

Conclusions: HepaRG-based 2D directed-nanopatterning coupled with CellProfiler analysis represents proof-of-concept for developing prototype LiverBiochips, as a convenient HTScompatible method for assessing multiparametric cell descriptors for drug development. P333 HUMAN HEPATIC HepaRG CO-CULTURE MODEL AS A SENSITIVE AND NON-INVASIVE TOXICOLOGICAL PLATFORM USING ECIS (ELECTRICAL CELL-SUBSTRATE IMPEDANCE SENSING) BIOSENSORS W. Gamal1 , P. Treskes2 , C. Chesne3 , J.N. Plevris2 , P.-O. Bagnaninchi1 , L.J. Nelson2 . 1 Centre for Biomedical Engineering, 2 Hepatology, University of Edinburgh, Edinburgh, United Kingdom; 3 Biopredic International, Rennes, France E-mail: [email protected] Background and Aims: A robust HTS-compatible multiparametric platform for assessing drug hepatotoxicity would provide a new paradigm for in vitro toxicology. Using the highly-differentiated human hepatic cell line HepaRG as a surrogate to primary human hepatocytes, our aim was to profile real-time changes in cellular impedance, Z , in response to paracetamol toxicity. Such dynamic screening allows capture and modeling of multiparametric, highcontent data using ECIS biosensors. Methods: HepaRG cells were cultured to confluence (terminallydifferentiated Hepatocyte:Cholangiocyte co-culture) on ECIS8W10+ and 8W1E-dd 8-well (250,000 cells/well) electrode arrays. Following CYP3A4 induction (day 8), with Rifampicin (24h), a paracetamol time-/dose-response [DMSO-vehicle as controls] was monitored with Impedance Z measurements taken every 180 sec


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POSTERS over a 500 Hz to 64 kHz frequency (f ) range, for 96h post-induction. Parallel ATP-depletion viability assays were performed (PromegaCellTiter© -Glo). ECIS-8W10+ had 40 electrodes/well (averaging Z signal over hundreds of cells). ECIS-8W1E-dd arrays had a range of electrode sizes ranging from 250um down to 20um (sample specific cell populations). Results: Z -time curves showed highly sensitive dose-related response to paracetamol over the critical 24h period [Figure 1], with decreasing Z values reflecting disruption of cell–cell/cell– substrate interactions (f = 4 kHz); whilst at f = 64 kHz, Z changes were indicative of metabolic derangement and loss of membrane integrity. HepaRG ATP content mirrored Z changes. Z -spectra modelling (utilizing ECIS algorithms) supported these observations, and provided further evidence of disruption of barrier function/celladhesion parameters.

oxidative stress. We demonstrated the presence of an extracellular fibrin network lining sinusoid spaces and the targeting of rcHIP/PAP to this inflammatory ECM during the course of experimental ALF in mice. The time window for the treatment of Fas-intoxicated mice (survival gain and ROS detoxification) with rcHIP/PAP corresponded to the time of occurrence of the fibrin-fibrinogen scaffold. We showed that the endogenous HIP/PAP gene was not yet activated at this time, emphasising the need to resort to rcHIP/PAP. Liver explants from patients that underwent transplantation for ALFs of various etiologies also displayed fibrin accumulation deposits in sinusoid spaces hinting at a probable clinical relevance of the HIP/PAP-fibrin interaction. Conclusions: rcHIP/PAP is an extracellular antioxidant with curative properties against ALF. P335 DYSFUNCTIONAL GUT MICROBIOTA IN ILEO-COLONIC MUCOSA AND STOOL IN ALCOHOLIC CIRRHOSIS IS ASSOCIATED WITH ENDOTOXEMIA: IMPLICATIONS FOR BACTERIAL TRANSLOCATION J.S. Bajaj1 , D. Heuman1 , P. Hylemon2 , M. Sikaroodi3 , J. Ridlon2 , W.M. Pandak1 , M. Schubert1 , M. Fuchs1 , M. White1 , K. Daita2 , P. Gillevet3 . 1 Gastroenterology, Hepatology and Nutrition, Virginia Commonwealth University and McGuire VA Medical Center, 2 Microbiology, Virginia Commonwealth University, Richmond, 3 Microbiome Analysis Center, George Mason University, Manassas, VA, United States E-mail: [email protected]

Figure 1. HepaRG paracetamol time- and dose-response measured by ECIS at 64 kHz.

Conclusions: Real-time high resolution tracking of morphological changes following hepatotoxic challenge was observed. The HepaRG-ECIS platform may offer a breakthrough technology for pre-clinical in vitro hepatotoxicology. P334 AN INFLAMMATORY EXTRACELLULAR MATRIX IS A MOLECULAR TARGET OF THE ROS SCAVENGER HIP/PAP N. Moniaux1,2,3 , M. Darnaud1,2 , K. Garbin1,2 , H. Yatri1,2 , 1,2,4 ´ , J. Faivre1,2,5 . 1 Unit A. Dos Santos1,2 , D. Samuel1,2,3 , C. Brechot opital 785, Inserm, 2 Univ Paris-Sud, 3 Centre Hepato-Biliaire, AP-HP, Hˆ Paul Brousse, Villejuif, 4 Institut Pasteur, Paris, 5 Service Hématologie et Biologie des Tumeurs, AP-HP, Hˆ opital Paul Brousse, Villejuif, France E-mail: [email protected] Background and Aims: The human recombinant HIP/PAP protein (rcHIP/PAP) has been shown to be a free radical scavenger that protects primary hepatocytes against several death effectors and significantly improved survival of mice bearing a chemically induced acute liver failure (ALF). It has been then tested in a phase 2 clinical trial in patients with ALF. We report on the mechanisms underlying this curative effect on inflammatory livers. Methods: To clarify this mechanism, we undertook a biochemical analysis of human hepatocyte primary cultures and mice bearing a Jo2-induced ALF treated with rcHIP/PAP. Results: In vitro, we showed that rcHIP/PAP localized within an extracellular matrix (ECM) network via a rcHIP/PAP-fibrin interaction, and that this ECM localization was required for rcHIP/PAP to detoxify ROS and protect human hepatocytes against S178

Background and Aims: Small bowel bacterial overgrowth/ translocation is linked to negative consequences in alcoholic cirrhosis but small bowel microbiota have not been studied in a culture-independent manner in cirrhosis. To evaluate dysbiosis (dysfunctional gut microbiota) in alcoholic cirrhosis compared to non-alcoholic cirrhotics and controls on stool, ileal and colonic biopsies. Methods: Cirrhotics and age-matched healthy controls underwent stool collection and serum endotoxin analysis. A subset underwent colonoscopy with ileal/colonic biopsies. Biopsies and stool were tested for percentage of autochthonous/beneficial bacterial families (Lachnospiraceae, Ruminococcaceae, ClostridialesXIV and Veillonellaceae) using multi-tagged pyrosequencing. Actively drinking alcoholic cirrhotics (Alc) were compared to non-alcoholic cirrhotics (NAlc) and controls. Results: 27 controls and 214 cirrhotics (20% Alc) were included (MELD alc 15 vs. NAlc 14, p = 0.81, Endotoxin Alc 1.5 vs. NAlc 0.6 vs. control 0.4, p = 0.02). Stool autochthonous taxa proportion was lower in Alc than NAlc/controls (26% vs. 38%vs.56%, p = 0.01). In the entire group, stool autochthonous bacteria proportion was negatively linked with endotoxemia (r-0.5, p = 0.014). 41 patients (10 Alc, 20 NAlc and 11 controls) underwent colonoscopy with rectal, descending, transverse, ascending colonic and ileal biopsies. There was a generalized significant reduction in the proportion of autochthonous bacteria in Alc compared to NAlc and controls at each level. However, relative abundances between various levels of the intestine within groups were similar (table). Table: Relative abundance of autochthonous bacteria

Stool Rectum Descending Transverse Ascending Ileum

Controls

Non-alcoholic cirrhosis

Alcoholic cirrhosis

56% 42% 43% 48% 45% 46%

38% 38% 42% 35% 33% 37%

26%*# 32%*# 29%* 34%# 30%# 32%

* significantly lower in alcohol compared to others; # significantly lower in cirrhotics compared to controls.


POSTERS Conclusions: We have found for the first time that stool dysbiosis reflects ileo-colonic mucosal dysbiosis in cirrhosis. This dysbiosis, characterized by a reduced beneficial bacterial relative abundance, is significantly more prevalent in actively drinking alcoholic cirrhotics, is linked with endotoxemia and could be associated with bacterial translocation in this population. P336 HEPATOCYTES IMPROVE THEIR DRUG METABOLIC ACTIVITY WITH THE PRESENCE OF ENDOTHELIAL CELLS IN AN IN VITRO HEPATIC CO-CULTURE MODEL OF ACETAMINOPHEN TOXICITY M. Navarro1 , L. Nelson1 , P. Treskes1 , P. Hayes1 , K. Samuel2 , J. Plevris1 . 1 Department of Hepatology, 2 SNBTS, University of Edinburgh, Edinburgh, United Kingdom E-mail: [email protected] Background and Aims: Hepatocyte cell lines are used in vitro to predict human hepatotoxicity and to investigative cell signaling in drug metabolism. However, in vivo, endothelial cells also facilitate drug deposition/clearance. We have therefore developed an in vitro hepatic model, incorporating human hepatic cells (HepG2/C3A) and human umbilical vein endothelial cells (HUVEC) to investigate the role of endothelial cells in drug metabolism. Methods: HepG2/C3A cells and HUVEC were cultured separately or co-cultured at a ratio of 1:1. After 2 days, cultures were exposed to 0mM, 5mM, 10mM, and 20mM of acetaminophen for 24h. CYP3A4 activity was assessed using P450-Glo™ CYP3A4 Assay with Luciferin-IPA (Promega), primary antibody (CYP3A4) and albumin synthesis with albumin Blue 580 Fluorescence Assay. Cell viability was measured using CellTiter-Glo Luminescent Cell Viability assay (Promega). Results: Co-Culture with HUVEC induced CYP3A4 activity in the absence of acetaminophen, which was further increased in cells exposed to 5mM and 10mM APAP. Albumin synthesis by HepG2/C3A cells was also increased on day 3. Further, HepG2/C3A cell survival following exposure to APAP overdose (10mM) of acetaminophen led to a significant reduction in cellular ATP

content associated with cell death and depletion of intracellular glutathione and total nitric oxide. By contrast, HepG2/C3A cells co-cultured with HUVEC showed a significantly higher ATP content when exposed to >10mM of acetaminophen with a relative increase in nitric oxide and glutathione. Conclusions: In this study, we have demonstrated an interaction between hepatocytes and endothelial cells in response to the hepatotoxic drug acetaminophen. In the presence of HUVEC, cell viability was maintained and synthesis of total nitric oxide and glutathione was increased in response to toxic insult. The application of this hepatoprotective effect of endothelial cells in vivo could enhance the integrity of the hepatic sinusoid endothelium, in addition to administration of N-acetyl-L-cysteine in acetaminophen poisoning. P338 ROLE OF ACID SPHINGOMYELINASE IN APAP-INDUCED LIVER INJURY A. Baulies1,2 , S. Nunez ˜ 1,2 , V. Ribas1,2 , L. Martinez1,2 , N. Kaplowitz3 , 1,2 1 ´ . Cell Death and C. Garc´ıa-Ruiz1,2 , J.C. Fernandez-Checa 2 Proliferation, IIBB-CSIC, Liver Unit, Hospital Clinic, IDIBAPS and CIBEREHD, Barcelona, Spain; 3 USC Research Center for Liver Diseases, Keck School of Medicine, Los Angeles, CA, United States E-mail: [email protected] Background and Aims: Acetaminophen (APAP) is a pain reliever that can cause liver necrosis and even acute liver failure in case of overdose. Recent studies show that activation of autophagy protects against APAP hepatotoxicity. Acid sphingomyelinase (ASMase) deficiency triggers a lysosomal storage disorder characterized by lysosomal cholesterol loading that compromises autophagy. Thus, we analyzed the role of ASMase in APAP-induced liver injury. Methods: Liver damage, histological profile and mitochondrial function were examined in ASMase−/− mice and their littermates following APAP treatment (300 mg/kg). Survival was examined in fasted animals following a lethal dose of APAP (500 mg/kg). Mitophagy was analyzed by confocal imaging with Lysotracker and DsRed-stained mitochondria. ASMase+/+ PMH were treated with U18666A, an intracellular cholesterol transport inhibitor, and ASMase−/− PMH with 25-hydroxycholesterol to diminish lysosomal cholesterol content. In all cases, cell viability was analyzed following APAP treatment. Results: Liver injury examined by serum ALT levels and histology was greater in ASMase−/− mice following APAP treatment compared to ASMase+/+ . This outcome paralleled the lower survival rate of the null mice 48 h after a lethal dose of APAP. ASMase−/− PMH cell viability after APAP treatment was significantly lower. Lysotraker and DsRed mitochondria colocalization was lower in ASMase−/− PMH following APAP treatment, indicating defective mitophagy. ASMase+/+ PMH pre-treated with U18666A showed enhanced susceptibility to APAP, while 25-hydroxycholesterol pretreatment in ASMase−/− PMH significantly protected against APAP induced cell death. Conclusions: Lack of ASMase enhances liver susceptibility to APAP treatment due to an impairment of autophagy as a consequence of the lysosomal cholesterol accumulation.


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POSTERS P339 PON1 DIRECTED RECALIBRATION OF MACROPHAGE LIPID HOMEOSTASIS LEADS TO REVERSAL OF CIRCULATING FOAMY PHENOTYPE AND PREDICTS OUTCOME IN ACUTE ON CHRONIC LIVER FAILURE (ACLF) PATIENTS J.S. Maras1 , S. Das2 , S.S. Jaswal2 , R. Mahiwal3 , C. Kumar3 , D. Kumar2 , H.H.C. Gawda4 , A. Rastogi5 , N.T. Pati2 , S. Sharma2 , A. Pandey4,6 , S.K. Sarin3 . 1 Institute of Liver & Biliary Sciences, 2 Department of Research, 3 Hepatology, Institute of Liver & Biliary Sciences, New Delhi, 4 Institute of Bioinformatics, Bangalore, 5 Department of Pathology, Institute of Liver & Biliary Sciences, New Delhi, India; 6 Departments of Biological Chemistry, Pathology, Oncology, and Surgery, McKusick-Nathans Institute of Genetic Medicine, Institute of Basic Biomedical Sciences, Section of Hepatology and Liver Transplantation, Johns Hopkins University School of Medicine, Baltimore, MD, United States E-mail: [email protected] Background and Aims: Acute on chronic liver failure is a rapidly progressive severe illness with a potential of reversibility. Lipid metabolism and transport is dysregulated in ACLF and foamy degeneration in severe alcoholics is correlated with mortality. Whether circulating proteins in ACLF potentiate alteration in macrophage lipid-homeostasis, phenotype or functional characteristics is not known. Aim: To determine the mechanism involved in dysregulation of lipid-homeostasis in macrophages in patients with ACLF due to alcohol. Methods: Quantitative protein expression profiling was validated using ELISA, on ACLF (n = 40), compensated cirrhosis (n = 20) and healthy-controls (n = 20). Combinatorial recombinant protein treatment [CRPT] on rat-BMDM/THP1 cells with conditional media (ACLF/CLD/HC-plasma) was assessed for lipid-homeostasis by qPCR/immuno-fluorescence/histochemistry for phenotypic/ functional characterization. Results: 120 proteins (>2 folds) were identified. ELISA-validation documented significant reduction of proteins involved in lipidhomeostasis (Pon1, ApoB, ApoA1, ApoA2, ApoC1, ApoC3 and ApoD) in ACLF [p≤0.05]. Significantly reduced levels of Pon1, ApoB and concomitant increase of ox-LDL was correlated with ACLF-nonsurvivors. BMDM/THP1 on ACLF condition-media documented increased nile-red-staining and altered expression of CD36, SRB1, FAS, SREBP1, TNF-alpha, IL-6, ABCA1,G1, LDLR, LXRb , EPO as compared to CLD and HC condition-media [>2 folds]. CRPT of [PON1, HDL, and ApoA1] on BMDM/THP1 reversed nile red staining and gene expression profile of macrophages. Circulating Pon1 (40%). ALP remission was defined as either normalization of ALP (for patients with baseline ALP 1.67–2.8×ULN)or an ALP less than 1.67×ULN (for patients with baseline ALP >2.8×ULN). Other endpoints included change in European Liver Fibrosis (ELF) scores and serum bile acids. Results: 20 patients were treated; 19 completed the Wk28 visit. One patient was discontinued as PBC was not confirmed. 95% of patients were female and median duration of disease was 3.2 yrs (range: 0.9, 13.2); 13 (65%) had baseline ALP >3×ULN. Baseline ELF scores indicated that patients had moderate to severe liver fibrosis (median score 9.8). No patients achieved ALP response or ALP remission at Wk12/Wk28. Median percent reduction from baseline in ALP concentration was 8.37% at Wk12 and 12.07% at Wk28. 13 patients (68%) had a reduction of ≥15% in ALP at either Wk20/Wk28 (Figure). There was a small decrease in median ELF score from baseline to Wk28. 6 patients (30%) had a decrease in ELF of ≥0.5 and 2 patients (10%) had increase of ELF score of ≥0.5 points. Total serum bile acid concentrations at baseline were highly elevated (median 43.32 mmol/L) with a decrease in concentration from baseline to Wk28 of 8.84 mmol/L (IQ Range: −18.925; 1.760 mmol/L). UST was well-tolerated with no SAEs, serious infections, and discontinuations due to AE through Wk28.

P368 COGNITIVE AND EMOTIONAL PROCESSING NETWORKS ARE IMPACTED BY SENSORY INPUT TO THE THALAMUS IN PATIENTS WITH PRIMARY BILIARY CIRRHOSIS V. Mosher1,2 , M. Swain3 , R.P. Myers3 , G. MacQueen4,5,6 , B. Goodyear2,5,7 . 1 Medical Sciences, 2 Seaman Family MR Research Centre, University of Calgary, 3 Liver Unit, Calgary Division of Gastroenterology and Hepatology, 4 Mathison Centre for Mental Health Research & Education, 5 Hotchkiss Brain Institute, 6 Psychiatry, 7 Radiology, University of Calgary, Calgary, AB, Canada E-mail: [email protected] Background and Aims: Behavioural symptoms including fatigue and cognitive dysfunction are common in patients with primary biliary cirrhosis (PBC) and suggest an alteration in brain function. To better understand the link between behaviour and brain function in PBC patients, we used resting-state functional magnetic resonance imaging (rs-fMRI) to determine if functional connections associated with emotional and cognitive processing in the brain are altered in PBC patients. Methods: Six PBC patients (none with advanced liver fibrosis), and six age/sex-matched healthy volunteers, underwent rs-fMRI. Brain maps of connection strength with the amygdalae were determined using time series analysis. These maps were compared between groups, with disease symptom severity (assessed using the PBC-40 questionnaire) as a covariate. Results: Compared to healthy controls, PBC patients exhibited reduced connection strength with the amygdalae to the ventral posterolateral thalamus, which receives sensory input from the body, and brain regions associated with high-emotion word processing, including the right lingual gyrus, right anterior cingulate cortex, and middle temporal gyri. PBC symptom severity (reflected by greater PBC-40 scores) was associated with reduced connection strength to the amygdalae within the insula (associated with body awareness and emotional processing), and the left inferior frontal gyrus (associated with language).

Figure: Brain regions exhibiting reduced functional connection strength with the amygdalae, in PBC patients relative to healthy volunteers (blueto-light blue), and regions where connection strength decreases with increased PBC symptom severity as measured by the PBC-40 (red-toyellow) (expressed as Z-score).

Conclusions: UST did not materially change serum ALP in patients with advanced, UDCA unresponsive, PBC.

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Conclusions: Our results suggest that PBC patients exhibit alterations of specific emotional and cognitive processing brain networks; an effect occurring via the ventral posterolateral thalamus. Importantly, a number of these alterations are linked to more severe behavioural symptoms in PBC patients. These findings link behavioural symptoms in PBC patients to altered brain function.


POSTERS P369 EXTRAHEPATIC MANIFESTATIONS OF ATP-BINDING CASSETTE TRANSPORTER B4 (ABCB4) DEFICIENCY DETECTED IN A SYSTEMATIC PHENOMICS PLATFORM K. Hochrath1 , M. Krawczyk1 , M. Hrabˇe de Angelis2,3,4 , F. Lammert1 . 1 Department of Medicine II, Saarland University Medical Center, Homburg, 2 German Research Centre for Environmental Health, German Mouse Clinic, Helmholtz Zentrum M¨ unchen, Munich, 3 Technische Universit¨ at M¨ unchen, Chair of Experimental Genetics, Freising-Weihenstephan, 4 Member of German Center for Diabetes Research (DZD), Neuherberg, Germany E-mail: [email protected] Background and Aims: The ABCB4 gene encodes the hepatocanalicular phospholipid floppase, which translocates phosphatidylcholine from liver into bile. Abcb4 knockout (Abcb4−/− ) mice develop chronic cholangitis and biliary fibrosis similar to primary sclerosing cholangitis in humans (Hepatololgy 2004;39:117–28, Gastroenterology 2004;127:261–74). To date, the interactions of these diseases with extrahepatic organs are not fully defined. With the aim to characterize the full spectrum of systemic effects of ABCB4 deficiency, we performed comprehensive whole body phenomics of Abcb4−/− mice. Methods: Standardized phenotyping screens (Nat Methods 2005;2:403–4) were performed in 20 Abcb4−/− mice on two different genetic backgrounds (FVB/N and BALB/c) as well as 40 control animals. These screens included phenotyping of dysmorphology, behavior, neurology, eye, nociception, energy-metabolism, clinical chemistry, immunology, allergy, steroid metabolism, metabolomics, cardiovascular function, lung function, and general pathology. Results: Genotype-specific differences were found in multiple screens. In particular, metabolomic analyses revealed a significant (p < 0.05) reduction of 23 of 163 (15%) analyzed lipids in knockout mice on both backgrounds. The immunology screen showed increased CD4+ T-lymphocyte counts (BALB/c: 5.34±0.47 vs. 4.54±0.56; p < 0.001) in Abcb4−/− mice, whereas the frequencies of granulocytes (BALB/c) and CD19+ B-cells (FVB/N) were decreased. Additionally, laser interference biometry determined enlarged axial eye lengths in Abcb4−/− mice as compared to controls (BALB/c: 3.626±0.034 vs. 3.577±0.017 mm; p < 0.001). No differences were observed in energy-metabolism, cardiovascular function, and nociception tests. Conclusions: Whole body phenotyping of Abcb4−/− mice illustrates the extrahepatic implications of a liver-specific transporter defect. This study may underscore the links between hepatic phenotypes and extrahepatic symptoms in chronic cholestasis. P370 CHOLESTATIC Abcb4-DEFICIENT MICE REVEAL DEPRESSION/ANXIETY-LIKE BEHAVIOR 2 K. Hochrath1 , S.M. Holter ¨ , L. Garrett2 , V. Gailus-Durner2 , H. Fuchs2 , 2,3,4 M. Hrabe´ de Angelis , W. Wurst5 , F. Lammert1 , C.S. Stokes1 . 1 Department of Medicine II, Saarland University Medical Center, Homburg, 2 German Research Centre for Environmental Health, German Mouse Clinic, Helmholtz Zentrum M¨ unchen, Munich, 3 Technische Universit¨ at M¨ unchen, Chair of Experimental Genetics, Freising-Weihenstephan, 4 Member of German Center for Diabetes at M¨ unchen, Chair Research (DZD), Neuherberg, 5 Technische Universit¨ of Developmental Genetics, Freising-Weihenstephan, Germany E-mail: [email protected] Background and Aims: Although depression is commonly associated with chronic liver diseases, its causality and functional mechanisms remain largely unknown (Psychosomatics 2013;54:52–9). Quantitative phenotypes are linked to psychiatric disorders, thus genetic studies in mice might help identify modifier genes that influence behavior. Our aim was to characterize the behavioral phenotype in the ABCB4-deficient (Abcb4−/− ) mouse, an established model for chronic cholestatic liver disease.

Methods: We evaluated spontaneous exploratory drive, reactivity to novelty, and emotionality with the open field (OF) test, in Abcb4−/− and BALB/cJ wild-type mice (N≥18). Specifically, time courses of distance travelled, speed, time spent in the center and rearing frequencies were noted. Additionally, sensorimotor gating by prepulse inhibition (PPI) at different intensities was assessed. Results: Abcb4−/− mice showed no overt behavioral abnormalities, constant weight gain, and normal reproduction rates. However, OF tests revealed genotype-dependent effects (p < 0.001) with consistently decreased locomotor activity in Abcb4−/− mice, characterized by reduced speed of movement and increased central zone avoidance. Furthermore, we detected decreased rearing activity for Abcb4−/− mice as compared to controls (47.8±8.4 vs. 99.0±8.6 [events], p < 0.001). PPI revealed a deficit at 69 dB intensity in female Abcb4−/− mice only. Conclusions: Abcb4−/− mice show greater anxiety behavior and reduced exploratory activity compared to controls. Given that ABCB4 is exclusively present in liver, the locomotor behavioral alterations are most likely due to neurochemical changes, e.g. reduced serum vitamin D levels, as recently observed in Abcb4−/− mice (BONE 2013;55:501–11). This preclinical model resembles the link between chronic liver diseases and increased stress susceptibility observed in humans. P371 PRIMARY SCLEROSING CHOLANGITIS-INFLAMMATORY BOWEL DISEASE IS ASSOCIATED WITH AN INCREASED FREQUENCY OF POST-TRANSPLANT COLONIC LYMPHOMA P.J. Trivedi1,2 , T. Wright1 , J. Robinson1 , K.-K. Li1,2 , D. Burns3 , B.K. Gunson1 , D.H. Adams1,2 , J. Ferguson2 , G.M. Hirschfield1,2 . 1 NIHR Biomedical Research Unit and Centre for Liver Research, University of Birmingham, 2 Liver Unit, Queen Elizabeth Hospital Birmingham, 3 School of Cancer Sciences, University of Birmingham, Birmingham, United Kingdom E-mail: [email protected] Background and Aims: Post-transplant lymphoproliferative disease (PTLD) is a recognised complication of liver transplantation (LT). Although small intestinal involvement is common, factors predisposing to colonic disease are not well characterised. Methods: A case-note review of all patients undergoing liver transplantation (1982–2013) was performed, and subsequently cross-referenced with an institutional lymphoma database comprising all biopsy-proven PTLD cases to date. Putative risk factors for development of colonic lymphoma were analysed using SPSSv21. Results: Over a 31-year adult ‘first liver’ transplant experience (No. of recipients=2872), 72 cases of post-LT lymphoproliferative disease were identified and most commonly observed in the context of primary biliary cirrhosis (PBC; n = 20) and primary sclerosing cholangtitis (PSC; n = 14). Overall, intestinal involvement was observed in 18/72 patients, representing predominantly small bowel disease (n = 12). Colonic lymphoma occurred only in individuals transplanted for PSC (n = 6; median 5.0yrs posttransplant; IQR: 3.2–11.7), all of who had underlying colitis. In all cases, disease was a diffuse large B-cell lymphoma; however, only 2 patients had EBV-(LMP)-positive tumours, of which only 1 a detectable serum EBV-titre (qPCR). There were no significant associations with age at transplantation, male gender, treatment with azathioprine or tacrolimus, duration of azathioprine or calcineurin inhibitor exposure, or onset of colitis post-LT (P=n.s.). Only one patient (each) with PTLD occurring in the context of cardiothoracic (n = 6) and renal (n = 32) transplantation developed large bowel disease, and under these circumstances was part of a disseminated lymphomatous process. Conclusions: PSC/colitis is associated with development of colonic lymphoma post-LT. Additional risk factors have yet to be identified.


S191

POSTERS P372 APOPTOSIS INDUCED BY BILIRUBIN AND LITHOCHOLIC ACID IN HUMAN OSTEOBLASTS IS NEUTRALIZED BY URSODEOXYCHOLIC ACID 4 S. Ruiz-Gaspa` 1,2 , M. Dubreuil2,3 , N. Guanabens ˜ , A. Combalia5 , on Biomédica P. Peris4 , A. Monegal4 , A. Pares6 . 1 Centro de Investigaci´ en Red en Enfermedades Hep´ aticas y Digestivas (CIBERehd), 2 Liver Unit, Digestive Diseases Institute, Hospital Cl´ınic, IDIBAPS, University on Biomédica en Red en of Barcelona, 3 Centro de Investigaci´ Enfermedades Hep´ aticas y Digestivas, 4 Rheumatology, 5 Department of Orthopedics, Metabolic Bone Diseases Unit, Hospital Cl´ınic, University of Barcelona, 6 Liver Unit, Hospital Cl´ınic, University of Barcelona, IDIBABPS, CIBERehd, Barcelona, Spain E-mail: [email protected] Background and Aims: Low bone turnover osteoporosis is common in cholestatic diseases. Ursodeoxycholic acid (UDCA) counteracts the damaging effects of bilirubin or lithocholic acid (LCA) on osteoblast viability, proliferation and mineralization. UDCA is antiapoptotic in various cell lines, but this effect in bone cells is unknown. Therefore, the consequences of bilirubin and LCA on apoptosis, and if UDCA has antiapoptotic effects have been assessed on osteoblasts. Methods: Human osteoblasts (hOB), and osteosarcoma cell line (Saos-2) were treated with camptothecin as a proapoptotic agent, and UDCA, LCA and bilirubin. Apoptosis was determined by DNA fragmentation, flow cytometry, caspase-3 activity, and expression of proapoptotic (Bcl-2-associated X protein BAX), and antiapoptotic (BCL2 and BCL2-like 1 protein, BCL2L) genes. Results: Both LCA (10 mM) and bilirubin (50 mM) induced apoptosis as indicated by DNA fragmentation (4.7 and 3.7 fold, respectively, p < 0.001), caspase-3 activity and flow cytometry in Saos-2 and hOB. UDCA (10 mM) reduced the apoptotic effects of camptothecin (0.5 mM) by 61%, (p < 0.001), and counteracted the apoptotic effects of LCA and bilirubin determined by DNA fragmentation (56% and 60%, respectively, p < 0.001), cytometry and caspase-3 activity in Saos-2, with lower effects in hOB. UDCA (10 mM) downregulated BAX (75%), and upregulated BCL2L (10-fold, p < 0.01) genes, and neutralized BAX upregulation (p < 0.01) and BCL2L downregulation (p < 0.01) induced by LCA and bilirubin. Conclusions: Bilirubin and LCA induce apoptosis in osteoblastic cells. UDCA counteracts the apoptotic consequences of these two substances and therefore, it may have further beneficial effects on the decreased bone formation in the cholestasis. P373 SERUM METABOLOMIC PROFILING IN PATIENTS WITH CHOLESTATIC PRURITUS. EFFECTS OF ALBUMIN DIALYSIS 2 A. Pares1 , M. Perez-Cormenzana ´ , A. D´ıaz1 , R. Mayo2 , A. Castro2 , 1 1 A. Mas . Liver Unit, Hospital Cl´ınic, CIBERehd, IDIBAPS, University of Barcelona, Barcelona, 2 OWL, Bilbao, Spain E-mail: [email protected] Background and Aims: This study assesses changes in the serum metabolomic profile before and after treatment with albumin dialysis using the molecular adsorbents recirculating system (MARS), to identify metabolites potentially associated with the pathogenesis of pruritus. Methods: Samples from sera before and after MARS and from albumin dialyzate were taken from 9 patients (7 women) with cholestasis and resistant pruritus. Metabolite extraction was accomplished by fractionating the samples into pools of species with similar physicochemical properties, and three different profiles were used: a) polar lipids, non-esterified fatty acids, and bile acids; b) amino acids, and c) apolar lipids. The analyses were performed by UPLC-ESI-QTOF-MS, random forests and multivariate and univariate analysis. Results: More than 470 metabolites were identified. Most metabolites decreased in sera after MARS and the differences S192

were particularly significant for sterols, N-acyl ethanolamines, 1-ether,2-acylglycerophosphoetanolamine and free sphingoid bases. A significant decrease of primary bile acids (p = 0.04) mainly taurine and glycine conjugates, and secondary bile acids, were found after MARS. The amino acid profile did not change, except for a significant decrease in cystine, taurine and tyrosine, and an increase in glutamine. A significant reduction of pregnenolone sulfate, androsterone sulphate isomer, and some glycerophospholipids and kynurenine were also found. Four of these serum metabolites and bile acids were particularly high in the albumin dialysate. Conclusions: Albumin dialysis is associated with a decrease of circulating metabolites especially phospholipids, primary bile acids and sterols. This metabolomic analysis identifies a panel of biomarkers that could participate into the pathogenesis of cholestatic pruritus. P374 LONG-TERM TREATMENT OF PRIMARY BILIARY CIRRHOSIS WITH THE FXR AGONIST OBETICHOLIC ACID SHOWS DURABLE EFFICACY K. Kowdley1 , G. Hirschfield2 , R. Chapman3 , C. Vincent4 , D. Jones5 , A. Pares6 , V. Luketic7 , S. Gordon8 , R. Pencek9 , T. Marmon9 , R. Hooshmand-Rad9 . 1 Virginia Mason Medical Center, Seattle, WA, United States; 2 University of Birmingham, Birmingham, 3 University of Oxford, Oxford, United Kingdom; 4 University of Montreal, Montreal, QC, Canada; 5 University of Newcastle, Newcastle, United Kingdom; 6 University of Barcelona, Barcelona, Spain; 7 Virginia Commonwealth University, Richmond, VA, 8 Henry Ford Health System, Detroit, MI, 9 Intercept Pharmaceuticals, San Diego, CA, United States E-mail: [email protected] Background and Aims: Obeticholic acid (OCA), 6-ethyl chenodeoxycholic acid (CDCA), is derived from CDCA and shows ~100× greater farnesoid X receptor (FXR) agonist activity than CDCA, the natural human ligand. OCA is being developed for the treatment of primary biliary cirrhosis (PBC). In a double blind, placebo controlled study OCA 10 mg and 50 mg, given as monotherapy, showed highly significant reductions in alkaline phosphatase (ALP) and other biochemical analytes, compared with placebo. Pruritus was the main adverse event (AE) in the DB phase (placebo: 30%, OCA 10 mg: 70%, 50 mg: 94%; Kowdley, J. Hepatology 2011; 54: S13). An open label, long-term study extension followed. Methods: During the ongoing extension, subjects initiated OCA at 10 mg once daily and titrated to 50 mg based on response. Ursodeoxycholic acid (UDCA) was added in 11 subjects. Results: Subjects (N = 28): mean age 58yrs; female: 78%; Caucasian: 96%. Baseline: ALP: 442±275U/L; bilirubin: 4.6±3.2 mmol/L; GGT: 460±318U/L; ALT: 91±61U/L; AST: 72±39 U/L. Median (IQR) exposure was 3.2 yrs (2.6–3.6); 9 subjects terminated early. Changes in liver chemistry are shown in the table. Pruritus was the most common AE (89%). Normalized exposure to pruritus (days/year of OCA) was comparable between the first year and overall but severity tended to decrease with continued treatment. Table: Changes in liver chemistry from baseline after 2 and 3 years Percentage change, mean (SE)

At 2 years All Subjects (n = 23) No UDCA use (n = 15) At 3 years All Subjects (n = 15) No UDCA use (n = 10)

ALP

GGT

ALT

−31% (9)* −29% (11)*

−48% (10)* −38% (8)* −39% (14)* −26% (11)*

−32% (12)* −48% (11)* −41% (8)* −33% (14)* −46% (12)* −41% (9)*

AST

Bilirubin

−22% (8)* −11% (9)

−17% (7)* −7% (9)

−27% (8)* −22% (9)*

−17% (10) −8% (12)

*p < 0.05 change from baseline.

Conclusions: PBC is a rare, chronic cholestatic liver disease with persistent significant unmet need. In this study long-term OCA


POSTERS treatment maintained a durable improvement in ALP and other liver chemistry. Pruritus, while prevalent appeared to diminish in severity with continued therapy. P375 SORTILIN DEFICIENCY ABOLISHES DUCTULAR REACTION AND REDUCES HEPATOCYTE APOPTOSIS AND LIVER FIBROSIS IN BILE DUCT LIGATION-INDUCED CHOLESTASIS I. Zvibel1 , E. Hubel1 , J. Dranoff2 , W.J. Park3 , E. Lavoie2 , N. Erez1 , S. Fishman1 , R. Bruck1 . 1 Research Center for Digestive Tract and Liver Diseases, Tel Aviv, Israel; 2 University of Arkansas for Medical Sciences, Little Rock, AR, United States; 3 Weizman Institute, Rehovoth, Israel E-mail: [email protected] Background and Aims: Sortilin is a recently described trafficking molecule directing newly synthesized molecules from the transGolgi network to secretory pathways or to the cell surface. One of the enzymes trafficked by sortilin and catalyzing sphingomyelin turnover to ceramide is acidic sphingomyelinase (ASM). ASM has been shown to be involved in hepatic fibrosis and hepatic stellate cells activation therefore, we have investigated the role played by sortilin deficiency in the development of liver fibrosis. Methods: Liver fibrosis was induced by bile duct ligation (BDL) or by 10 wks administration of CCl4 to WT and sortilin−/− mice. Results: Sortilin−/− mice displayed strongly attenuated hepatic fibrosis in bile duct ligation (BDL)-induced fibrosis, but not in CCl4 induced fibrosis, as shown by Sirius red staining for collagen and by mRNA levels of collagen I, aSMA, TGFb and TIMP1. At early stages after BDL, sortilin−/− mice had reduced ductular reaction, accompanied by lower levels of MCP-1, IL-6, TNFa and CXCL1 and reduced infiltration of Ly6G-positive neutrophils in the peri-ductular areas. Serum levels of IL-6 and its downstream signal pSTAT3 were also reduced in sortilin−/− mice. Cholangiocyte proliferation, as demonstrated by Ki67 staining, peaked in WT mice 7d after BDL, but was strongly attenuated in sortilin−/− mice. Sortilin−/− mice showed attenuated liver damage, less bile infarcts and reduced hepatocyte apoptosis. Livers from sortilin−/− mice had less ASM activity 7 and 14 days after BDL, compared to WT. Conclusions: Sortilin deficiency leads to reduced ductular reaction and reduced fibrosis after BDL. P376 LOSS OF KERATIN 19 PREDISPOSES TO THE DEVELOPMENT OF CHOLESTATIC LIVER DISEASE VIA IMPAIRED OVAL CELL ACTIVATION 1 Y. Chen1 , N. Guldiken ¨ , M. Spurny2 , J. Hayback ¨ 3 , M. Pollheimer4 , P. Fickert4 , N. Gassler5 , M.K. Jeon5 , C. Trautwein1 , P. Strnad1 . 1 IZKF and Department of Internal Medicine III, University Hospital Aachen, Aachen, 2 Department of Internal Medicine I, University Medical Center Ulm, Ulm, Germany; 3 Institute of Pathology, 4 Institute of Pathology and Department of Internal Medicine, Medical University Graz, Graz, Austria; 5 Institute of Pathology, University Hospital Aachen, Aachen, Germany E-mail: [email protected] Background and Aims: Keratins (K) are established cytoprotective proteins. Among them, K19 is highly expressed in ductal epithelia and hepatic progenitor cells. K19 expression associates with a poor prognosis in hepatocellular carcinoma and K19 variants are found in patients with primary biliary cirrhosis. Methods: To study the role of K19 in biliary system and hepatic progenitor cells, K19 wild-type (WT) and knockout (KO) mice were subjected to two oval cell proliferation models, i.e. feeding with 0.1% 3,5-diethoxycarbonyl, 1,4-dihydrocollidine (DDC) or with choline-deficient, ethionine-supplemented (CDE) diet for four and two weeks, respectively. Supplementation with 1% cholic acid (CA) for 1 week and common bile duct ligation (CBDL) were used to analyze the impact of K19 on biliary system.

Results: In untreated animals, loss of K19 led to re-distribution of keratin network in cholangiocytes. After DDC feeding, K19-KO mice exhibited (compared to WT) (i) increased cholestatic parameters (bilirubin, ALP; serum bile acids); (ii) lower ductular proliferation and lower amount of oval cells; (iii) impaired Notch 2 signaling in biliary cells; (iv) lower biliary fibrosis. After CDE diet, K19 deficiency resulted in a stronger hepatic injury and lower amount of oval cells. On the other hand, loss of K19 affects neither bile acid excretion nor bile acid-induced hepatotoxicity (CA model). Moreover, neither differences in liver injury nor biliary fibrosis development were noted 5 and 21 days after CBDL. Conclusions: Our results indicate that K19 plays an important role in oval cell proliferation and related liver regeneration while it is dispensable in biliary system. P377 THE FXR AGONIST OBETICHOLIC ACID IMPROVES A TRANSPLANTFREE SURVIVAL-PROVEN BIOCHEMICAL RESPONSE CRITERION IN PLACEBO CONTROLLED PRIMARY BILIARY CIRRHOSIS STUDIES V. Luketic1 , S.G. Gordon2 , C. Vincent3 , R. Chapman4 , M. Mayo5 , K. Kowdley6 , G. Hirschfield7 , A. Mason8 , K. Lindor9 , L. Macconell10 , T. Marmon10 , R. Hooshmand-Rad10 , D. Shapiro10 . 1 Virginia Commonwealth University, Richmond, VA, 2 Henry Ford Health System, Detroit, MI, United States; 3 University of Montreal, Montreal, QC, Canada; 4 University of Oxford, Oxford, United Kingdom; 5 University of Texas Southwestern Medical Center, Dallas, TX, 6 Virginia Mason Medical Center, Seattle, WA, United States; 7 University of Birmingham, Birmingham, United Kingdom; 8 University of Alberta, Edmonton, AB, Canada; 9 Arizona State University, Tempe, AZ, 10 Intercept Pharmaceuticals, San Diego, CA, United States E-mail: [email protected] Background and Aims: Obeticholic acid (OCA), a novel derivative of chenodeoxycholic acid and potent farnesoid-X receptor agonist, is being developed for the treatment of Primary Biliary Cirrhosis (PBC). The prevalence and slow progression of PBC makes clinical outcome (liver transplant or death) studies extremely difficult. The Global PBC Study Group has showed that alkaline phosphatase (ALP) and bilirubin correlated with transplant-free survival in PBC patients (±UDCA) (Lammers Hepatology 2013). Specific response criteria (ALP < 1.67×ULN, ≥15% reduction and bilirubin ≤ ULN) are being studied as the primary endpoint in the Phase 3 PBC OCA International Study of Efficacy (POISE). Methods: Two 12-week, double-blind, placebo-controlled Phase 2 trials evaluated the safety and efficacy of OCA in PBC patients with persistently high ALP (≥1.5–10×ULN) and bilirubin < 2×ULN, with or without UDCA (n = 224). All OCA doses showed highly significant decreases in ALP and bilirubin versus placebo. OCA was generally well-tolerated. Dose-related pruritus was common. Pooled data were retrospectively evaluated using the POISE response criteria. Subjects above the POISE entry criteria were analyzed for response (n = 189). Results: Patient demographics were similar across groups (mean age = 55 years; female [≥90%]; White [≥95%]). Significantly more OCA-treated patients met the POISE response criteria versus placebo, in both the OCA monotherapy and UDCA-background therapy studies. Table: POISE PBC response criteria† , % (95% CI) Treatment

Pooled Overall p-value*** Monotherapy p-value*** In combination with UDCA p-value***

Placebo (N = 53)

10 mg OCA (N = 46)

25 mg OCA (N = 39)

50 mg OCA (N = 48)

Total OCA (N = 133)

8 (2–18) − 5 (0–24) − 9 (2–25) −

41 (27–57) 10,.000 IU/ml exhibited PPR, whereas only 3/21 (14.3%) women with HBV-DNA 2,000 and the third with HBV-DNA of 1,850 IU/ml) exhibited PPR. Conclusions: In the present study, PPR occurred in ~30% of HBeAgnegative women, and all events were recorded in the first semester after delivery. Pre-partum HBV-DNA levels >10,000 IU/ml was a predictor of PPR. P665 SOLUBLE ST2 PLASMA CONCENTRATIONS PREDICT MORTALITY IN HBV-RELATED ACUTE-ON-CHRONIC LIVER FAILURE Z.-Y. Lei, Z.-S. Mo, X.-H. Li, Z.-B. Wu, K. Wang, D.-Y. Xie, Z.-L. Gao. The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou, China E-mail: [email protected] Background and Aims: Soluble ST2 (sST2) is a decoy receptor for IL-33, which acts as an alarmin of inflammation. This study mainly focused on the plasma levels and clinical significance of sST2 in HBV-related acute-on-chronic liver failure (HBV-ACLF). Methods: 48 patients admitting to our hospital were defined as HBV-ACLF base on the standard: total bilirubin (TB) ≥85 umol/L and prothrombin time international normalized ratio (INR) ≥1.5. Patients surviving shorter or longer than 180 days since admission, called death (n = 11) and survival (n = 35) group respectively. Blood was obtained weekly from baseline to week 3. 12 patients with chronic hepatitis B (CHB) and 16 healthy people (HC) served as controls. ELISA was used to detect sST2 levels in plasma, while Luminex was used to detect IL-33, IL-10, TNFa and IFN-g levels simultaneously. Results: The level of sST2 in HBV-ACLF at baseline was remarkably higher than that in controls (P < 0.001), while TNFa, IL-10 levels were only higher than that in HC. After treatment, sST2 level progressively increased in death group, but decreased in survival group, there were significant differences between two groups at S290

week 2 (P = 0.002) and week 3 (P < 0.001). The concentration of sST2 at week 3 was related to TB (r = 0.751, P < 0.001), INR (r = 0.629, P < 0.001) and Model for End-stage Liver Disease (MELD) scores (r = 0.735, P < 0.001). Kaplan–Meier survival analysis showed that higher sST2 concentration predicted lower survival rate (Figure 1).

Figure 1. Kaplan–Meier survival curve showing survival probability of 48 patients with HBV-ACLF who were stratified into two groups according to median plasma sST2 concentration at week 3.

Conclusions: Our data subjected that sST2 level correlated with disease severity and mortality of HBV-ACLF. To some extent, it may help to predict the poor prognosis for HBV-ACLF. P666 PREDICTORS OF SEVERE HEPATIC FIBROSIS IN TREATMENTNAIVE GENOTYPE B OR C INFECTED E ANTIGEN-POSITIVE AND -NEGATIVE CHRONIC HEPATITIS B PATIENTS W.-P. Su, C.-Y. Peng, H.-C. Lai, C.-H. Lin, P.-H. Chuang, S.-H. Chen. Division of Hepatogastroenterology, Department of Internal Medicine, China Medical University Hospital, Taichung, Taiwan E-mail: [email protected] Background and Aims: We investigated the predictors of severe hepatic fibrosis and their predictive values in a consecutive cohort of treatment-naïve, HBeAg-positive and -negative chronic hepatitis B (CHB) patients. Methods: A total of 409 treatment-naïve CHB patients (204 HBeAgpositive and 205 HBeAg-negative) were enrolled. The liver histology was scored using the METAVIR system. Predictive factors including age, gender, BMI, platelet, hemoglobin, white count, ALT, genotype, and HBV DNA and quantitative hepatitis B surface antigen (qHBsAg) levels were collected at the time of liver biopsy. The cut-off values for the predictors were determined by AUROC analysis. Results: Independent factors associated with severe fibrosis (F3–4) were genotype C infection (OR = 3.549, 95% CI: 1.738– 7.245, P = 0.0005), platelet count (OR = 0.981, 95% CI: 0.974–0.988, P < 0.0001) and qHBsAg (OR = 0.538, 95% CI: 0.336–0.864, P = 0.0102) in HBeAg-positive patients, and were BMI (OR = 1.153, 95% CI: 1.035–1.285, P = 0.0095), platelet count (OR = 0.975, 95% CI: 0.966– 0.984, P < 0.0001) and HBV DNA (OR = 0.663, 95% CI: 0.537–0.818, P = 0.0001) in HBeAg-negative patients. Combining the cut-off of 50 years old. The clinical course was particularly severe in patients with underlying chronic liver disease. Conclusions: HEV RNA testing allows for the diagnosis of acute hepatitis E. Consistent with previous reports, men above 50 constituted the majority of patients presenting with symptomatic autochthonous infection in Switzerland. P702 HEV INFECTION IS A FREQUENT CAUSE (20%) OF ELEVATED TRANSAMINASES IN PATIENTS WITH INFLAMMATORY RHEUMATIC DISEASES: RESULTS OF A FRENCH MONOCENTRIC PROSPECTIVE STUDY S. Fournier1 , M. Guillaume1 , C. Luxembourger2 , F. Abravanel3 , H. Bagheri4 , M. Wargny4 , C. Bureau1 , J.-P. Vinel1 , A. Cantagrel2 , 1 1 . Hepatology and Gastroenterology, 2 Rheumatology, J.-M. Peron ´ 3 4 Virology, Pharmacology, Toulouse University Hospital, Toulouse, France E-mail: [email protected] Background and Aims: Elevated liver enzymes in patients treated in rhumatology are often attributed to a drug toxicity. Prevalence of hepatitis E in this population is not known. The aim of this study was to prospectively describe the causes of elevated liver enzymes in these patients. Methods: This is a french monocentric prospective study. All patients followed in the Rheumatology Department of Toulouse University Hospital from Feb 2012 to Mar 2013 and presenting ALTs >2ULN were included. HEV infection was diagnosed by detection of anti HEV IgM (Wantai, China) or HEV RNA in serum. Results: 82 patients were included. The main causes of elevated transaminases were drug toxicity (37.8%), NAFLD (18.2%) and acute hepatitis E (7.3% of the whole population and 20.6% in the sub-group of patients with inflammatory rheumatic disease). Seroprevalence (IgG) was 51.8%. The 6 patients who developed acute hepatitis E had positive HEV IgM and 4 were viremic. Median age was 53 years old (25–70). One patient had jaundice (bilirubin 65 mmol/L). Median ALT was 410 UI/L (121–3430). All of them were taking immunosuppressing drugs (methotrexate in 5, antiTNFa in 3). Patients HEV infected were more often treated by methotrexate than others (p < 0.001). None had severe liver failure or chronic infection. Three viremic patients have been treated with ribavirin until RNA undetectability. None had viral relapse. Conclusions: Acute HEV infection is a frequent cause of elevated ALTs in the rhumatologic population in particularly in those treated with immunosuppressors. None had chronic infection. The place of ribavirin therapy needs to be further defined.

P703 THE GLOBAL BURDEN OF HEPATITIS A VIRUS IN 1990 AND 2005 D.B. Rein1 , G. Stevens2 , A. Flaxman3 , J.S. Wittenborn1 , N. Timothy4 , S.Z. Wiktor4 , S.T. Wiersma5 . 1 Public Health Department, NORC at the University of Chicago, Atlanta, GA, United States; 2 Department of Health Statistics and Information Systems, World Health Organization, Geneva, Switzerland; 3 Institute for Health Metrics and Evaluation, University of Washington, Seattle, WA, United States; 4 Global Hepatitis Programme, World Health Organization, Geneva, Switzerland; 5 U.S. Centerers for Disease Control and Prevention, Atlanta, GA, United States E-mail: [email protected] Background and Aims: We estimated the infections and morbidity from hepatitis A virus (HAV) in 1990 and 2005 using a simulation modeling approach. Methods: We estimated regional seroprevalence of anti-HAV using the Global Burden of Disease Study’s DISMOD-MR model to analyze abstracted data from 605 published studies. We converted seroprevalence into incidence using 3 models of HAV fit to the characteristics of high-income, low-income, and economically transitioning regions between 1990 and 2005. We used published parameters to convert incident infections into asymptomatic, symptomatic without death, or fatal disease outcomes. Results: For 1990, we estimated 99.4 million (95% credible interval [Cr.I.] 88.7–109.6) incident HAV infections, resulting in 28.7 (95% Cr.I. 1.0–71.4) million symptomatic cases, and 22,498 (95% Cr.I. 562–47,039) deaths. For 2005, we estimated 108.7 (95% Cr.I. 98.8– 118.1) million incident HAV infections, resulting in 32.9 (95% Cr.I. 1.1–80.6) million symptomatic cases, and 59,601 (95% Cr.I. 1,284– 108,341) deaths. Between 1990 and 2005, incidence decreased and the average age of infection increased. Table: Estimated hepatitis A virus outcomes: 1990 & 2005 Measure

1990 (95% Cr.I.)

2005 (95% Cr.I.)

Change

Global Population (billions) Incident infections (millions) Symptomatic cases (millions) Deaths

5.306 99.4 (88.7–109.6) 28.7 (1.0–71.4) 22,498 (562–47,039)

6.506 108.7 (98.9–118.1) 32.9 (1.1–80.6) 56,601 (1,284–108,341)

22.6% 9.4% 14.6% 264.9%

Conclusions: We estimate an increase in HAV infections that is consistent with population growth but also a disproportionate increase in HAV deaths. Our model indicates this is the result of infections occurring at older age when HAV infection is more likely to be severe. Although our estimates contain a great deal of uncertainty, if these results are confirmed by country surveillance data, routine infant vaccination against HAV should be included as part of a comprehensive plan for the prevention and control of viral hepatitis. P704 CLINICAL COURSE AND TREATMENT OF CHRONIC HEPATITIS E INFECTION IN PATIENTS AFTER SOLID ORGAN TRANSPLANTATION 1 E. Halilbasic1 , L. Kazemi-Shirazi1 , K. Staufer1 , P. Munda1 , C.J. Muller ¨ , H. Holzmann2 , P. Jaksch3 , W. Klepetko3 , M. Trauner1 , H. Hofer1 . 1 Department of Internal Medicine III, Division of Gastroenterology and Hepatology, 2 Department of Virology, 3 Department of Thoracic Surgery, Medical University of Vienna, Vienna, Austria E-mail: [email protected] Background and Aims: Chronic hepatitis E virus (HEV) infection is an emerging cause of liver injury in solid organ transplant (SOT) recipients. Due to immunosuppression management of HEV infection in SOT patients can be challenging. We report clinical course and treatment outcome of chronic HEV infection in SOT patients from an Austrian tertiary center. Methods: Five SOT patients (1 kidney, 4 lung, 4 males, median age 58 [24–65] years) tested positive for serum HEV-RNA. All patients underwent a liver biopsy. Immunosuppression included


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POSTERS prednisolone with tacrolimus (N = 4) or cyclosporine (N = 1) with or without mycophenolate mofetil. The 4 lung transplant recipients were treated with ribavirin (RBV) and followed up for up to 19 months. Results: HEV infection became apparent 4 [1–9] (median [range]) months after transplantation. All patients had elevated liver enzymes at diagnosis of HEV infection (mean ALT: 79 [62–110] U/l; [range]) and were infected with HEV genotype 3. Liver biopsy revealed presence of cirrhosis in 3 patients (Metavir fibrosis stage 1 and 2 in the remaining two). During RBV treatment ALT values normalized in all, but HEV-RNA became undetectable in only 2 patients. One Patient achieved a SVR, the other relapsed. The kidney-transplanted patient presented with liver failure and died prior to initiation of ribavirin treatment. Conclusions: Chronic HEV infection may have a rapid fibrosis progression and is an important cause of liver related morbidity in patients after transplantation. Treatment with ribavirin results in viral suppression and biochemical normalization, although it might not be successful in all patients on the long run. P705 FULL GENOME SEQUENCING AND CHARACTERIZATION OF A NEW HAV STRAIN TYPE 1B CIRCULATING IN ITALY C. Argentini1 , S. Catone1 , N. Coppola2 , U. Villano3 , S. Taffon3 , E. Sagnelli2 , M. Pisaturo2 , C. Sagnelli2 , S. Vella1 , D. Genovese1 . 1 Department of Pharmacs, Istituto Superiore di Sanit` a, Roma, 2 Dipartimento di Salute Mentale e Medicina Pubblica, Sezione di Malattie Infettive, Seconda Universit` a di Napoli, Napoli, 3 Department of Microbiology, Istituto Superiore di Sanit` a, Roma, Italy E-mail: [email protected] Background and Aims: Between 1997 and 2004, in Apulia and Campania a series of outbreaks have been described and circulation of genotype 1b was for the first time reported in Italy. To characterize the genotype 1b circulating in Italy. We have sequenced 6 isolates collected in Campania in 2001. Methods: We collected 19 HAV isolates (2001–2004). Virus genomes were amplified by PCR and sequenced. The sequence were analysed by means of MEGA 5.2, MODELTEST and BEAST to infere phylogenenis and virus evolution. Results: Based on 2A-VP1 region, 11 out of 19 isolates were classified as genotype 1b. We obtained full genome molecular sequences of three 1b isolates (S13, S17 and S18) and partial sequences (95% of total genome) of other 3 isolates. The comparison of genotype 1b completely sequenced demonstrate that these HAV isolates belong to a newly described strain, highly homogeneus (homology among isolates was 99.3% in nucleotide analyses and 99.4% in protein analyses) but strongly separated to previously described strain. The bayesan analyses showed that this strain probably arised between 60 and 80 years ago, but has been described in Italy only in the early 2000. Conclusions: This new HAV 1b strain is associated in a series of outbreacks in southern of Italy. We have demonstrated that is only distantly related to previously characterized genotype 1b strain escribed in Australia, USA and Africa. The application of phylogeny techniques allows us to suppose the emergence of this strain during HAV evolution and its circulation in Italy.

P706 RISK FACTORS OF CHRONIC KIDNEY DISEASE IN HIV-NEGATIVE PATIENTS WITH CHRONIC HEPATITIS B INFECTION: A FRENCH NATIONWIDE HOSPITAL COHORT STUDY 3 V. Mallet1,2 , S. Thiebaut ´ , A. Trylesinski4 , M. Schwarzinger3 . 1 Université Paris Descartes, Assistance Publique – Hˆ opitaux de Paris, Inserm, 2 Lingha ‘Links for Global Health Assessment’, Paris Biotech Santé, 3 THEN ‘Translational Health Economics Network’, Paris, France; 4 Novartis Pharma AG, Basel, Switzerland E-mail: [email protected] Background and Aims: Chronic kidney disease (CKD) is associated with chronic hepatitis B virus (CHB) infection at all liver disease stages. Our aim was to determine the risk factors of CKD in CHB patients before decompensated cirrhosis. Methods: We conducted a retrospective, longitudinal analysis of the 2010–2012 French National Hospital database. CHB adult patients were identified by ICD-10 medical coding (B18.0/B18.1). CKD was defined by a glomerular filtration rate below 60 mL/min, dialysis or kidney transplant. Exclusion criteria involved: HIV co-infection; decompensated cirrhosis; and, at cohort inception, CKD or organ transplant. Risk factors of CKD were estimated according to obesity status in proportional hazards models using age as the time-scale and death as a competing risk. Results: Of 23,161 CHB patients (male: 52.2%; mean (SD) age: 45.1 (16.7) years), 546 (2.4%) progressed to CKD and 559 (2.4%) died from non-liver causes during 42,974 patient-years. Obesity status defined two at-risk groups of CKD since 135/2,156 (6.3%) obese (BMI ≥ 30 kg/m2 ) patients progressed to CKD as compared to 411/21,105 (2.0%) non-obese patients (p < 0.0001). The incidence of CKD increased sharply with age and male gender in both groups (Figure). Independent risk factors of CKD were similar in obese/nonobese CHB patients: co-infection with delta virus (HR = 2.18/1.33), high-blood pressure (HR = 2.00/3.74), insulin-dependent diabetes (HR = 6.00/4.90), non-insulin-dependent diabetes (HR = 3.64/2.34), and a history of acute kidney injury (HR = 1.91/2.08). About 50% obese and 25% non-obese CHB patients had at least one risk factor of CKD before decompensated cirrhosis.

Figure: Incidence of CKD in French CHB HIV-negative patients according to obesity status.

Conclusions: Obesity is a major risk factor of CKD in CHB patients.

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POSTERS P707 CHRONIC KIDNEY DISEASE INCREASES NON-LIVER-RELATED MORTALITY IN HIV-NEGATIVE PATIENTS WITH CHRONIC HEPATITIS B INFECTION: A FRENCH NATIONWIDE HOSPITAL COHORT STUDY 3 V. Mallet1,2 , S. Thiebaut ´ , A. Trylesinski4 , M. Schwarzinger3 . 1 Université Paris Descartes, Assistance Publique – Hˆ opitaux de Paris, Inserm, 2 Lingha ‘Links for Global Health Assessment’, Paris Biotech Santé, 3 THEN ‘Translational Health Economics Network’, Paris, France; 4 Novartis Pharma AG, Basel, Switzerland E-mail: [email protected] Background and Aims: Chronic kidney disease (CKD) is associated with chronic hepatitis B virus (CHB) infection at all liver disease stages. Our aim was to determine the role of CKD in non-liverrelated mortality in CHB patients. Methods: We conducted a retrospective, longitudinal analysis of the 2010–2012 French National Hospital database. CHB adult patients were identified by ICD-10 medical coding (B18.0/B18.1). CKD was defined by a glomerular filtration rate below 60 mL/min, dialysis or kidney transplant. Exclusion criteria involved: HIV co-infection; and, at cohort inception, CKD, organ transplant, or end stage liver disease (ESLD) including hepatocellular carcinoma. Prognosis factors of non-liver-related death were estimated in a proportional hazards model stratified by gender and obesity status, using age as the time-scale and ESLD as a competing risk. Results: Of 25,881 CHB patients (male: 54.8%; mean (SD) age: 46.3 (16.9) years), 2,602 (10.1%) progressed to ESLD (case-fatality rate: 32.2%) and 628 (2.4%) died from non-liver-related causes during 48,653 patient-years. CKD occurred in 51/628 (8.2%) patients who died from non-liver-related causes, 150/2,602 (5.8%) patients before ESLD, and 507/22,651 (2.2%) patients otherwise (p < 0.001). Independent prognostic factors of non-liver-related mortality were: chronic alcoholism (HR = 1.39, p < 0.001), progression to CKD (HR = 4.24, p < 0.0001), and other major Charlson Comorbidities (Index = 1: HR = 4.90; Index = 2: HR = 9.48; Index ≥ 3: HR = 24.02, p < 0.0001). Independent risk factors of ESLD were: chronic alcoholism (HR = 3.31, p < 0.0001), co-infection with delta virus (HR = 1.39, p < 0.0001) or chronic hepatitis C (HR = 1.59, p < 0.0001), and insulin-dependent diabetes (HR = 1.27, p < 0.0001). Conclusions: CKD is an independent prognostic factor of non-liverrelated mortality in CHB patients. P708 CHRONIC HEPATITIS E – THE SPECTRUM OF HISTOPATHOLOGY A. Beer1 , S. Pischke2 , P. Behrendt2 , J. Schlue2 , U. Drebber3 , H. Kreipe2 , H. Wedemeyer2 , M. Manns2 , H.P. Dienes1 . 1 University of Vienna, Vienna, Austria; 2 Hannover Medical School, Hannover, 3 University of Cologne, Cologne, Germany E-mail: [email protected] Background and Aims: Sporadic hepatitis E is an emerging indigenous disease in Europe induced by genotype III of the virus. Whereas in immunocompetent individuals it takes an acute selflimited course, under immunocompromised conditions chronic hepatitis may develop. Histopathology of acute hepatitits E has been published whereas histology of the chronic hepatitis E has not been evaluated. Methods: We collected liver biopsies from 14 patients (13 male, 1 female; age 10–66 years) who had undergone solid organ transplantation (4 liver, 5 kidney, 4 heart). One HIV negative patient had CD4 lymphocyte impairment. Biopsies were stained with H&E, Prussian Blue, van Gieson and PASD. Evaluation of histologic activity and fibrosis was performed applying Ishak score. Results: Biochemical data showed a considerable increase with ALT mean levels 326 U/L (35–1410), AST mean 442 U/L (25–2574), gGT 192 (42–606), bilirubin 36 mg/dl (9–146). Histopathology displayed typical features of chronic hepatitis of mild to moderate activity

(histological activity index of 8, mild fibrosis stage 2). In two biopsies a conspicuous central vein sclerosis and perisinusoidal fibrosis was present that first lead to misdiagnosis of NASH. The number of PMN leucocytes was increased. Bile ducts were involved with a mild reactive cholangitis. Significant cholestasis was absent. Conclusions: Chronic hepatitis E runs a similar course as hepatitis B and C. Histopathology shows a pattern of mild to moderate histologic activity with an increased presence of PMN leucocytes and reactive cholangitis. Fibrosis showed stage 2 in the patients studied with a pattern resembling NASH in some. P709 LIVER HISTOLOGICAL INFLAMMATION OF CHRONIC HEPATITIS B VIRUS CARRIERS WITH PERSISTENTLY NORMAL ALANINE TRANSAMINASE D. He, Q. Shang, Y. An, Q. Yi, Y. Zhang, M. Ding. The 88th Hospital of Chinese PLA, Tai’an, China E-mail: [email protected] Background and Aims: A new upper limits normal of ALT (30 U/L for males and 19 U/L for females, relative to the current 40 U/L) is recommended. We aimed to observe the difference of liver histological inflammation in chronic hepatitis B virus carriers with persistently normal ALT classified by the two standards. Methods: 326 chronic HBV carriers with persistently normal ALT (202 immune tolerance state and 124 inactive HBsAg carrier state) were divided into low ALT group (≤30 U/L for males and ≤19 U/L for females) and high ALT group (31–40 U/L for males and 20–40 U/L for females). According to Ishak scoring system and Metavir grading algorithm, liver histological inflammation were evaluated. Results: Significant differences of Ishak inflammation score were seen between low ALT group and high ALT group (immune tolerance state: p = 1.55×10−11 ; inactive HBsAg carrier state: p = 1.06×10−8 ). In low ALT group, the proportion of Metavir mild inflammation in immune tolerance state and inactive HBsAg carrier state was 100% and 96.8%, respectively. In high ALT group, the proportion was 63.8% and 50.8%, respectively. A significant difference of Metavir mild, moderate, and severe distribution was seen in age and sex for immune tolerance state and in sex for inactive HBsAg carrier state. Conclusions: In conclusions, there is a significant difference of liver histological inflammation between the two standards of ALT. The new standard should be better than the current one in evaluating the mild histological inflammation. Sex and age are associated with histological inflammation in high ALT group.

8B. VIRAL HEPATITIS: HEPATITIS C – CLINICAL (EXCEPT THERAPY) P710 EFFECT OF ALISPORIVIR (ALV) ON THE PHARMACOKINETICS (PK), SAFETY, AND TOLERABILITY OF METHADONE IN HEALTHY SUBJECTS AND OPIOID-DEPENDENT PATIENTS ON STABLE METHADONE MAINTENANCE THERAPY (MMT) S.J. Kovacs1 , J. Ke1 , A. Barve1 , L.R. Webster2 , Y. Cheng3 , J. Zhang1 , R. Maietta4 , G. Sunkara1 , D.S. Stein1 . 1 Novartis Institutes for BioMedical Research, East Hanover, NJ, 2 CRI Lifetree, Salt Lake City, UT, United States; 3 Beijing Novartis Pharma Co. Ltd, Shanghai, China; 4 Novartis Institutes for Biomedical Research, Cambridge, MA, United States E-mail: [email protected] Background and Aims: HCV-infected patients requiring MMT represent an important population with specific needs. Methadone, a substrate for CYP2B6, CYP2C9, CYP2C19, CYP2D6, CYP3A4, does not require active transport to cross membranes. ALV is an


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POSTERS irreversible CYP3A4 inhibitor, and inhibitor of multiple cellular uptake and efflux transporters. This study evaluated the interaction between ALV and methadone to assess potential clinical risks for methadone withdrawal or toxicity. Methods: Open-label screening (Part 1) and randomized, placebocontrolled, double blind (Part 2) investigation. Healthy subjects received a single 5 mg dose of methadone before and after ALV (Part 1). Then, opioid-dependent patients on stable MMT (≤150 mg daily) were randomized (2:1) to receive ALV 600 mg BID or placebo for 7 days (Part 2). Safety assessments included daily vitals and 12lead ECGs, laboratory tests, and continuous pulse oximetry. Blood samples were collected for R- and S-methadone concentrations prior to and after dose administration on D1 (before ALV) and D8. Non-compartmental PK analysis was performed. Results: 16 healthy and 22 MMT subjects (14 active; 8 placebo) were enrolled. Two healthy and 2 MMT subjects withdrew consent before study completion. Dizziness (37.5%), headache (18.8%), and nausea (18.8%) were the most commonly reported AEs among healthy subjects; somnolence (30.8% vs. 25.0%), headache (30.8% vs. 12.5%), nausea (15.4% vs. 0%), and tremor (15.4% vs. 0%) among MMT subjects receiving ALV vs. placebo. Table: Summaries of the dose-normalized methadone exposure parameters from Part 2 Analyte

Dose-normalized parameter

Methadone (D1)

ALV+methadone (D8)

Geometric mean ratios (90% CI)

R-methadone

AUC0–24,ss [(ng·h/mL)/mg]

60.4±25.2*

64.0±24.5

1.071 (0.958, 1.196)

S-methadone

Cmax,ss [(ng/mL)/mg] AUC0–24,ss [(ng·h/mL)/mg]

3.69±1.27 60.9±32.4

3.68±1.13 61.1±29.8

1.001 (0.904, 1.109) 1.014 (0.895, 1.150)

Cmax,ss [(ng/mL)/mg]

4.24±1.81

4.21±1.57

1.002 (0.889, 1.129)

*Arithmetic mean ± SD.

Conclusions: Alisporivir does not affect the in vivo disposition of R- or S-methadone suggesting there is no relevant interaction that would complicate its use in the treatment of opioid-dependent patients infected with HCV. P711 A NOVEL GENETIC MAKER TO IMPROVE THE PREDICTION OF HCV SPONTANEOUS CLEARANCE: POLYMORPHISMS CONSISTING OF (TA)N DINUCLEOTIDE REPEAT NEAR IL28B GENE M. Sugiyama1 , S. Hiramine2 , N. Furusyo2 , A. Ido3 , H. Tsubouchi3 , H. Watanabe4 , Y. Ueno4 , M. Korenaga1 , K. Murata1 , N. Masaki1 , J. Hayashi2 , M. Mizokami1 . 1 National Center for Global Health and Medicine, Ichikawa, 2 Kyushu University, Hakata, 3 Kagoshima University, Kagoshima, 4 Yamagata University, Yamagata, Japan E-mail: [email protected] Background and Aims: Recent GWAS revealed that SNPs around IL-28B were associated with spontaneous clearance (SC). However, the effect of IL28B SNPs was lower than the results observed in the study of SC. We previously reported that the length of the TA dinucleotide repeat in the promoter region of IL28B has a wide variation, and the transcriptional activity increased gradually in a TA repeat length-dependent manner. In the present study, we determined the length of the TA repeat to investigate the correlation to with SC. Methods: Total 2041 Japanese genomic samples were enrolled (274 with HCV spontaneous clearance, 457 with chronic HCV infection and 1310 healthy donors). IL28B SNPs and the length of TA repeat were genotyped using Invader assay and GeneMapper software, respectively. Results: A univariate analysis showed significant differences between SC and chronic infection in sex (men 51.4% vs. women 67.0%), age (68.1±11.1 vs. 64.0±10.8 years), IL28B (rs8099917) genotypes (TT 92.0% vs. non-TT 67.4%), and TA repeat number (≥12/12, 99.6% vs. 25 kg/m2 (r = −0.55, p = 0.01). Serum chemerin levels were negatively associated with fibrosis stage (r = −0.37, p = 0.01) and inflammatory grade (r = −0.30, p = 0.04), independently of BMI. Serum chemerin

level correlated, but surprisingly negatively, with chemerin (r = −0.43; p = 0.004). Expression of visfatin correlated positively with inflammatory and steatosis grade (r = 0.45, p = 0.02; r = 0.42, p = 0.04; respectively) only in patients with BMI < 25 kg/m2 , while chemerin negatively with steatosis grade in patient with BMI > 25 kg/m2 (r = −0.50, p = 0.01). No association between liver gene expression and liver fibrosis and HOMA-IR was found. Chemerin correlated positively with chemerin receptor (r = 0.41, p = 0.002). Conclusions: BMI is a predictor of some adipokines liver gene expression. Only serum chemerin is associated, although negatively, with its liver gene expression. Serum chemerin is associated with fibrosis and inflammatory process. Liver gene expression of some adipokines may influence steatosis and inflammatory process. P715 A STEADY INCREASE IN PROPORTION OF HCV INFECTIONS DUE TO GENOTYPE 3 IN THE CANADIAN PROVINCE OF ALBERTA FROM 1998 TO 2013: IMPLICATIONS FOR THERAPY S.D. Shafran1 , K.E. Doucette1 , J.W. Tang2 . 1 Medicine, 2 Medical Microbiology and Immunology, University of Alberta, Edmonton, AB, Canada E-mail: [email protected] Background and Aims: Genotyping of HCV is necessary to guide treatment. HCV has been genotyped in the Canadian province of Alberta (population ~4M) since 1998 exclusively by the Alberta Provincial Laboratory for Public Health. Methods: We reviewed all HCV genotypes performed in Alberta since 1998 in persons aged 20–49 years to evaluate the distribution of HCV genotypes over time. We selected “younger” adults since they are likely to be closer to the date of HCV acquisition and therefore more representative of HCV transmission epidemiology. Trend data were examined by the Cochran-Armitage method. Results: HCV genotyping was performed in 8145 unique patients aged 20 to 49 years from 1998 to 2013. There was a steady increase in the proportion of genotype 3 cases from 20.9% in 1998–2000 to 30.1% in 2011–2013, a 44% increase (p < 0.0001), and a steady decrease in the proportion of cases due to genotype 1 from 69.1% in 1998–2000 to 58.4% in 2011–2013 (p < 0.0001). The proportion of cases due to genotype 2 was stable at ~9%. Less than 3% of HCV genotypes were due to non-genotypes 1, 2 and 3. These are displayed in the figure.

Figure: HCV genotype distribution over time.

Conclusions: The significant increase in the proportion of HCV cases due to genotype 3 at the “expense” of genotype 1 cases has implications for treatment, since patients with genotype 3 HCV infection are treated with less expensive antiviral regimens with less intensive laboratory monitoring than patients with HCV genotype 1.


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POSTERS P716 PERSISTENCE OF HEPATITIS C VIRUS AFTER SPONTANEOUS RECOVERY FROM HEPATITIS C A.Y. Chen1 , M. Hoare2 , G.J.M. Alexander3 , T.I. Michalak1 . 1 Molecular Virology and Hepatology Research Group, Memorial University, St. John’s, NL, Canada; 2 Cancer Research UK Cambridge Institute, 3 Medicine, University of Cambridge, Cambridge, United Kingdom E-mail: [email protected] Background and Aims: HCV causes chronic hepatitis C, while spontaneous recovery from infection is relatively infrequent. HCV expression after spontaneous resolution has rarely been investigated. We analyzed, using high sensitivity HCV RNA detection approaches, plasma and PBMC from individuals with a past episode of hepatitis C and repeatedly normal liver enzymes and undetectable HCV RNA by conventional testing. Methods: Parallel plasma and PBMC from 23 non-viraemic individuals by standard testing, followed for 0.3–14.3 (mean 6.3) years prior to initial collection, were examined. From 9 of them, additional samples were obtained 4.5–7.2 (mean 5.9) years later. RNA was extracted from 500 ml plasma and, if HCV negative, from ~10 ml after ultracentrifugation, and from PBMC cultured with PHA or PHA-PWM-IL2-IL4 (JGV2005;86:657). HCV RNA positive and negative strands were detected by RT-PCR/NAH with a sensitivity of 0.05). In FISSION, patients receiving PEG-IFN+RBV had substantially lower EQ5D scores compared to SOF+RBV (0.650±0.224 vs. 0.737±0.251, p = 0.041). Patients receiving PEG-IFN+SOF+RBV (NEUTRINO) had a major decrease in their EQ5D scores during treatment (from 0.793±0.219 to 0.645±0.213, p < 0.0001) similar to scores observed in patients receiving PEG-IFN+RBV (FISSION) (from 0.771±0.236 to 0.650±0.224, p < 0.0001). After 12 weeks of follow-up, patients with SVR-12 (FUSION) showed significant improvement in EQ5D scores (+0.043 to the baseline, p = 0.0127). In multivariate analyses, baseline depression, anxiety, fatigue, insomnia, treatment-related anemia and receiving IFN were predictors (p < 0.001) of lower EQ5D scores. Conclusions: This is the first study estimating EQ5D scores from SF-36 in CH-C. EQ5D scores are minimally impacted by IFN-free SOF regimens. These patients had significantly higher EQ5D scores than those receiving IFN-containing regimens.


POSTERS P719 INDEPENDENT PREDICTORS OF PATIENT-REPORTED OUTCOMES (PROS) AND QUALITY OF LIFE (QOL) IN CHRONIC HEPATITIS C PATIENTS RECEIVING INTERFERON (IFN)-FREE VERSUS IFN-CONTAINING REGIMENS WITH SOFOSBUVIR (SOF) Z.M. Younossi1,2 , M. Stepanova1,2 , F. Nader1 , D. Nelson3 , E. Lawitz4 , I.M. Jacobson5 , E. Gane6 , S.L. Hunt1,2 . 1 Center for Liver Diseases, Department of Medicine, Inova Fairfax Hospital, 2 Betty and Guy Beatty Center for Integrated Research, Inova Health System, Falls Church, VA, 3 University of Florida, Gainesville, FL, 4 Texas Liver Institute, University of Texas Health Science Center, San Antonio, TX, 5 Weill Cornell Medical College, New York, NY, United States; 6 Auckland City Hospital, Auckland, New Zealand E-mail: [email protected] Background and Aims: IFN causes substantial impairment of QoL. We compared PROs and QoL between IFN-free and IFN-containingSOF regimens. Methods: 525 patients receiving IFN+RBV+SOF (NEUTRINO) or RBV+SOF (FUSION) completed PRO questionnaires: CLDQ-HCV (QoL), FACIT-F (Fatigue) and WPAI-SHP (work productivity and activity). Subjects were blinded to virologic data. Results: Baseline depression/anxiety, insomnia and fatigue were similar between IFN-free or IFN-containing regimens (p > 0.05). By the end-of-treatment, decrements in PRO scores were higher in those receiving IFN-regimen: physical well-being (decrement = 18.9% IFN-based vs. 4.6% IFN-free, p < 0.0001), functional well-being (13.2% vs. 6.1%, p = 0.0002), fatigue (19.4% vs. 2.7%, p < 0.0001), totalFACIT-F (12.2% vs. 2.4%, p < 0.0001), all domains of CLDQ-HCV (9.5% to 19.3% vs. 0.3% to −3.3%, p < 0.05), work productivity (23.2% vs. 5.7%, p = 0.004) and activity impairment (22.2% vs. 6.5%, p = 0.002). In multivariate analysis, predictors of poor PROs at baseline were depression (beta = −9.4 to −13.6%, p < 0.05) and fatigue (beta = −12.2 to −22.5%, p < 0.05). At the end-of-treatment, predictors of poor PROs included receiving IFN (beta = −4.7 to −13.2%, p < 0.05), depression (beta = −6.4 to −6.5%, p < 0.05), fatigue (beta = −10.9 to −16.5%, p < 0.05) and female gender (beta = −7.8 to −9.4%, p < 0.05). By follow-up week 12, predictors of poor PROs included depression, anxiety, fatigue and cirrhosis (p-values 0.05). Compared to their baseline scores, patients receiving SOF+RBV experienced modest declines in some aspects of SF-36, CLDQ-HCV, fatigue and WPAI:SHP (p = 0.04 to 0.05). In patients achieving SVR-12 (regardless of the regimen), significant improvements were noted in General Health (p = 0.0004), CLDQ-HCV (P < 0.0001), Fatigue (p = 0.005), Emotional Well-Being (p=P < 0.0001) and Physical Component Summary Scores (p = 0.0022). Multivariate analyses showed that baseline depression, fatigue, insomnia and cirrhosis were the most consistent predictors of PRO impairment (all p < 0.05). Conclusions: PROs are minimally impacted by SOF+RBV regimens. An additional 12 weeks of treatment does not substantially add to the PRO burden. SVR-12 is associated with improvement in most PROs. Depression, fatigue, insomnia and cirrhosis are independent predictors of PROs. P721 INTERFERON-STIMULATED GENE EXPRESSION IS ASSOCIATED WITH TREATMENT RESPONSE TO FALDAPREVIR PLUS PEGYLATED INTERFERON a-2a AND RIBAVIRIN IN PATIENTS WITH CHRONIC HCV GENOTYPE-1 INFECTION Y. Datsenko1 , P. Baum1 , R. Schirmbeck2 , J.O. Stern3 , N. Sha3 , G.G. Steinmann1 . 1 Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, 2 Department of Internal Medicine I, Ulm University Hospital, Ulm, Germany; 3 Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT, United States E-mail: [email protected] Background and Aims: Interferon-stimulated gene (ISG) expression is associated with treatment response to pegylated interferon a-2a and ribavirin (PR). We investigated ISG expression before and during treatment with the NS3/4A protease inhibitor faldaprevir plus PR, and tested association of ISG expression with SVR. Methods: Treatment-naïve and treatment-experienced patients with HCV genotype-1 (GT-1) from the SILEN-C1 and 2 studies with available blood samples for RNA testing were included (N = 263). mRNA expression (TaqMan PCR) of various ISGs (N = 15; C1S, CCL8, CXCL10, DDX58, HESX1, IFI27, IFIT1, IFNA4, IRF3, MAVS, PKLR, RSAD2, SIGLEC1, TBX3, USP13) was measured at baseline, 4 hours, 4 days and 8 days post-treatment initiation in peripheral blood mononuclear cells (PBMCs). Genes associated with SVR (p ≤ 0.05) Gene

Timepoint Baseline

4 hours

Day 4

Day 8

IFNA4 IRF3 DDX58 RSAD2 IFI27

OR 1.128 (p < 0.05) OR 1.395 (p = 0.05) OR 1.278 (p < 0.05) n.s. n.s.

n.s. OR 0.749 (p < 0.05) n.s. n.s. n.s.

n.s. n.s. OR 1.160 (p < 0.05) OR 1.018 (p < 0.05) OR 1.003 (p < 0.05)

n.s. n.s. n.s. n.s. OR 1.003 (p < 0.05)

n.s., not significant; OR, odds ratio.

Results: The baseline characteristics of the 171 treatment-naïve and 92 treatment-experienced patients analysed were balanced and comparable with the overall population in SILEN-C1 and 2. All ISGs that showed an association with SVR (P≤0.05), including DDX58 and IFNA4, are presented in the table. SVR patients had lower baseline expression of C1S, CXCL10, DDX58 and IFNA4 and higher on-treatment induction of DDX58, HESX1, IFI27, IFIT1, PKLR


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POSTERS and RSAD2 compared with non-SVR patients (P≤0.05). Compared with treatment-experienced patients, treatment-naïve patients had lower baseline expression and higher on-treatment induction of the majority of the genes tested. Conclusions: The results of this study suggest that in addition to other baseline predictors (e.g. IL28B), baseline expression of the ISGs, IFNA4 and DDX58, in PBMCs can contribute to better prediction of SVR in HCV GT-1-infected patients treated with faldaprevir plus PR.

there was no association of IFNL3 and IFNL4 with SVR in patients with genotype 2 and 3 (p > 0.05). Conclusions: IFNL3 genotyping from serum was highly efficient and revealed concordant results using EDTA blood. In our study IFNL4(ss469415590) was highly associated with IFNL3-(rs12979860), thus we see no additional benefit for decision making of IFNL4(ss469415590) testing in Caucasian GT2/3 patients. IFNL3 is associated with RVR.

P722 PERFORMANCE AND VALUE OF IFN-LAMBDA3 AND IFN-LAMBDA4 GENOTYPING IN PATIENTS WITH CHRONIC HEPATITIS C (CHC) GENOTYPE 2/3 IN A REAL WORLD SETTING

P723 FALDAPREVIR PLUS PEGYLATED INTERFERON/RIBAVIRIN DID NOT INCREASE ANAEMIA COMPARED WITH PEGYLATED INTERFERON/RIBAVIRIN IN HCV GENOTYPE-1, TREATMENTNAIVE PATIENTS: POOLED ANALYSIS OF PHASE III STUDIES T. Asselah1 , D.M. Jensen2 , G.R. Foster3 , M.S. Sulkowski4 , D. Ouzan5 , L. Morano6 , R. Buynak7 , K. Agarwal8 , T. Hassanein9 , D. Forton10 , F. Negro11 , D. Genne´ 12 , K. Kaita13 , A. Maieron14 , L. Preotescu15 , 18 ´ , J.O. Stern19 , C. Sarrazin16 , E. Zehnter17 , M. Romero-Gomez 20 ¨ , A.-M. Quinson19 , M. Drulak19 , M. Garcia19 , F. Voss20 , W. Bocher 21 1 opital Beaujon, APHP, University Paris-Diderot and P. Ferenci . Hˆ INSERM CRB3, Clichy, France; 2 University of Chicago Medicine, Chicago, IL, United States; 3 Queen Mary, University of London, London, United Kingdom; 4 John Hopkins University School of Medicine, Baltimore, MD, United States; 5 Institut Arnault Tzanck, St Laurent du Var, France; 6 Hospital Meixoeiro, Vigo, Spain; 7 Northwest Indiana Center for Clinical Research, Valparaiso, IN, United States; 8 King’s College Hospital, London, United Kingdom; 9 Southern California Liver Centers, Coronado, CA, United States; 10 St George’s Hospital, London, United Kingdom; 11 University Hospital, Geneva, 12 Chaux de Fonds Hospital, Chaux de Fonds, Switzerland; 13 HSC University of Manitoba, Winnipeg, MB, Canada; 14 Elizabethan Hospital, Linz, Austria; 15 National Institute of Infectious Diseases ‘Prof Dr Matei Bals’, Bucharest, Romania; 16 J.W. Goethe University Hospital, Frankfurt, 17 Schwerpunktpraxis Hepatologie, Dortmund, Germany; 18 Valme University Hospital, Sevilla, Spain; 19 Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT, United States; 20 Boehringer Ingelheim Pharma GmbH & Co. KG, Ingelheim, Germany; 21 Medical University of Vienna, Vienna, Austria E-mail: [email protected]

5 S.B. Wiegand1,2 , B. Heidrich1,2 , S. Susser3 , A. Stoehr4 , K.H.W. Boker ¨ , N. Grigorian6 , R. Zachoval7 , R. Link8 , U. Naumann9 , C. John10 , A. Lohse11 , P. Malfertheiner12 , H. Carls13 , M.P. Manns1,2 , H. Wedemeyer1,2 , C. Sarrazin3 , M. Cornberg1,2 , for the HepNet Study Group. 1 Dept. of Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, 2 German Liver Foundation, HepNet Study-House, Hannover, 3 Dept. of Internal Medicine I, Johann Wolfgang Goethe University Medical Center, Frankfurt, 4 IFI Institute for Interdisciplinary Medicine, Asklepios Klinik St Georg, Hamburg, 5 Leberpraxis, Hannover, 6 Dept. of Internal Medicine II, Saarland University Medical Center, Homburg, 7 University of Munich, Munich, 8 St. Josefs Hospital, Offenburg, 9 Center for Addiction-Medicine, 10 Medical Practice, Berlin, 11 Hamburg Eppendorf, Hamburg, 12 Otto usseldorf, von Guericke University, Magdeburg, 13 Medical Practice, D¨ Germany E-mail: [email protected]

Background and Aims: SNPs near the interferon lambda 3 gene (IFNL3, IL28B) are predictors for SVR in patients with Chronic Hepatitis C GT1. Recently, a dinucleotide frame shift in ss469415590 was described, which generates IFNL4. In this study, we compared the role of IFNL4 variants with IFNL3-(rs12979860) and IFNL3(rs8099917) on response to PEG-IFN and Ribavirin in patients with Chronic Hepatitis C GT2/3. Methods: We recruited 1006 patients with Chronic Hepatitis C and GT2/3 in a large German registry. 959 started treatment with PEG-IFN and Ribavirin. We performed genotyping of IFNL3 (rs12979860, rs8099917; n = 641) and of IFNL4 (ss469415590; n = 631) from serum samples. Results: In 641 out of 695 patients (92%) serum samples results for genotyping were obtained. Concordance of genotyping in patients with available serum samples and EDTA blood samples (n = 219) was more than 96% for both IFNL3 SNPs. IFNL3(rs12979860) correlated with IFNL4: 99.2% of patients with IFNL3-(rs12979860)-CC were IFNL4-(ss469415590)-TT/TT. IFNL3(rs12979860)-CT was linked with IFNL4-(ss469415590)-TT/DG (98.0%) and IFNL3-(rs12979860)-TT was associated with IFNL4(ss469415590)-DG/DG (97.6%). Patients with the preferable IFNL3(rs12979860 and rs8099917) SNPs (CC vs. non-CC and TT vs. non-TT) had significantly higher RVR rates (p < 0.0001 for both). However,

Background and Aims: The advent of first-generation inhibitors have revolutionised HCV treatment, but anaemia and associated adverse events limit their use. In the Phase III STARTVerso1 and 2 trials, the potent HCV NS3/4A inhibitor faldaprevir plus pegylated interferon a-2a and ribavirin (PR) achieved high SVR rates in treatment-naïve patients with chronic HCV genotype-1 (GT-1) infection. We analysed pooled haematological data from STARTVerso1 and 2 to determine if faldaprevir increased PRassociated anaemia. Methods: Patients (N = 1309) received 24 weeks (W) or 48W PR plus: placebo for 24W; faldaprevir 120 mg QD for 12W or 24W; or faldaprevir 240 mg QD for 12W. Changes in haemoglobin, haptoglobin and reticulocyte levels were analysed.

Table (abstract P723): Change in haemoglobin from baseline

Haemoglobin levels, mean difference (±SD), g/dL Week 2 Week 4 Week 8 Week 12 Haptoglobin levels, mean maximum decrease (±SD), mg/dL Reticulocyte levels, mean maximum increase (±SD), %

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PR + Placebo

PR + Faldaprevir

(n = 264)

120 mg QD (n = 521)

240 mg QD (n = 524)

−1.1 (1.2) −2.1 (1.5) −2.6 (1.5) −2.8 (1.5) −35 (40) 1.9 (1.2)

−1.1 (1.2) −2.2 (1.5) −2.7 (1.4) −2.9 (1.4) −32 (43) 2.0 (1.7)

−0.8 (1.2) −1.9 (1.3) −2.5 (1.3) −2.9 (1.4) −27 (43) 1.7 (1.2)


POSTERS Results: Changes from baseline in haemoglobin, haptoglobin and reticulocyte levels are shown in the table. Of patients receiving 120 mg and 240 mg faldaprevir, 4% and 3%, respectively had haemoglobin ≤8.5 g/dL versus 3% for placebo. Of faldaprevir-treated patients, 27% in each arm had haemoglobin ≤10 g/dL versus 25% for placebo. 30%, 26% and 24% (placebo, faldaprevir 120 mg and faldaprevir 240 mg, respectively) of patients required ribavirin dose reductions and 3%, 4% and 3%, respectively, required erythropoiesisstimulating agents. 1% of patients in each arm required blood transfusions. Conclusions: Addition of faldaprevir to PR did not change the severity and time course of PR-related anaemia in HCV GT-1, treatment-naïve patients.

agents. The effect of ribavirin modifications on SVR12 rates is shown in the table. Conclusions: The proportion of patients who experienced haemoglobin ≤10 g/dL was similar in each of the treatment arms. Ribavirin dose reduction was the most common action taken in these patients and this did not appear to be associated with lower SVR12 rates compared with patients who had no change in ribavirin.

P724 VIROLOGICAL RESPONSE IN TREATMENT-NAIVE PATIENTS WITH CHRONIC HCV GENOTYPE-1 INFECTION RECEIVING FALDAPREVIR PLUS PEGYLATED INTERFERON a-2a AND RIBAVIRIN IS UNAFFECTED BY RIBAVIRIN DOSE REDUCTION T. Asselah1 , D.M. Jensen2 , G.R. Foster3 , M.S. Sulkowski4 , D. Ouzan5 , L. Morano6 , R. Buynak7 , K. Agarwal8 , T. Hassanein9 , D. Forton10 , M. Cho11 , D. Genne´ 12 , K. Kaita13 , A. Maieron14 , L. Preotescu15 , C. Sarrazin16 , E. Zehnter17 , A. Streinu-Cercel18 , J.O. Stern19 , Y. Datsenko20 , M. Drulak19 , F. Voss20 , A.-M. Quinson19 , P. Ferenci21 . 1 Hˆ opital Beaujon, APHP, University Paris-Diderot and INSERM CRB3, Clichy, France; 2 University of Chicago Medicine, Chicago, IL, United States; 3 Queen Mary, University of London, London, United Kingdom; 4 Johns Hopkins University School of Medicine, Baltimore, MD, United States; 5 Institut Arnault Tzanck, St Laurent du Var, France; 6 Hospital Meixoeiro, Vigo, Spain; 7 Northwest Indiana Center for Clinical Research, Valparaiso, IN, United States; 8 King’s College Hospital, London, United Kingdom; 9 Southern California Liver Centers, Coronado, CA, United States; 10 St George’s Hospital, London, United Kingdom; 11 Pusan National University, Yangsan Hospital, Busan, Korea, Republic of, 12 Chaux de Fonds Hospital, Chaux de Fonds, Switzerland; 13 HSC University of Manitoba, Winnipeg, MB, Canada; 14 Elizabethan Hospital, Linz, Austria; 15 Carol Davila University of Medicine and Pharmacy, Bucharest; National Institute for Infectious Diseases ‘Prof. Dr. Matei Bals’, Bucharest, Romania; 16 J.W. Goethe University Hospital, Frankfurt, 17 Schwerpunktpraxis Hepatologie, Dortmund, Germany; 18 Carol Davila University of Medicine and Pharmacy Carol Davila University of Medicine and Pharmacy, Bucharest; National Institute for Infectious Diseases ‘Prof. Dr. Matei Bals’, Bucharest, Romania; 19 Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT, United States; 20 Boehringer Ingelheim Pharma GmbH & Co. KG, Ingelheim, Germany; 21 Medical University of Vienna, Vienna, Austria E-mail: [email protected]

Ribavirin reduction only 38/64 (59) Ribavirin interruption 5/19 (26) No change 88/181 (49)

Background and Aims: STARTVerso1 and 2 evaluated faldaprevir plus pegylated interferon a-2a and ribavirin (PR) in treatmentnaïve patients with chronic HCV genotype-1 infection. This posthoc analysis explored the impact of ribavirin dose reduction or interruption on virological response in these trials. Methods: Patients (N = 1309) received 24 weeks (W) or 48W PR plus: placebo for 24W; faldaprevir 120 mg QD for 12W or 24W; faldaprevir 240 mg QD for 12W. Ribavirin dosing was weight-based: 1000 mg or 1200 mg BID (60–≤80 kg or >80 kg, respectively). Patients were categorised as having ribavirin ‘dose reduction’, ‘dose interruption’ or ‘no change’. Results: In placebo, faldaprevir 120-mg and 240-mg arms, 25%, 27% and 27% of patients, respectively had haemoglobin ≤10 g/dL. Overall, 60% of patients with haemoglobin ≤10 g/dL had a ribavirin dose reduction only, 13% had RBV interruption and 27% had no change. Among patients with a reduction or interruption in ribavirin, 20% prematurely discontinued ribavirin treatment. Overall, 1% required a blood transfusion and 4% received erythropoiesis-stimulating

Table: Effect of ribavirin dose modifications on SVR12 SVR12 based on ribavirin modifications∗† , n/N (%) Placebo

∗

Faldaprevir 120 mg

Faldaprevir 240 mg

88/114 (77) 18/28 (64) 276/379 (73)

85/109 (78) 32/41 (78) 261/374 (70)

Ribavirin dose modification for any reason is the denominator. analysis. ITT, intent to treat.

†

ITT

P725 IMPACT OF THE rs8099917 SNP ON SVR12 IN PATIENTS WITH HCV GT-1 INFECTION TREATED WITH FALDAPREVIR, DELEOBUVIR AND RIBAVIRIN T. Asselah1 , S. Zeuzem2 , V. Soriano3 , J.-P. Bronowicki4 , A.W. Lohse5 , 6 , M. Schuchmann7 , M. Bourliere ` 8 , M. Buti9 , B. Mullhaupt ¨ S. Roberts10 , E. Gane11 , F. Voss12 , J.-P. Gallivan12 , I. Herichova13 , opital Beaujon, APHP, University P. Baum14 , F. Mensa15 . 1 Hˆ Paris-Diderot and INSERM CRB 3, Clichy, France; 2 J. W. Goethe University Hospital, Frankfurt, Germany; 3 Hospital Carlos III, Madrid, opital de Brabois, Vandoeuvre, France; 5 University Hospital Spain; 4 Hˆ Hamburg-Eppendorf, Hamburg, Germany; 6 University Hospital of Zurich, Zurich, Switzerland; 7 University Hospital Mainz, Mainz, opital Saint Joseph, Marseille, France; 9 Hospital Vall Germany; 8 Hˆ d’Hebron, Barcelona, Spain; 10 Alfred Hospital, Melbourne, VIC, Australia; 11 Auckland Clinical Studies, Auckland, New Zealand; 12 Boehringer Ingelheim Pharma GmbH & Co. KG, Ingelheim, Germany; 13 Boehringer Ingelheim Pharma RCV GmbH & Co KG, Vienna, Austria; 14 Boehringer Ingelheim Pharma GmbH & Co. KG, Biberach, Germany; 15 Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT, United States E-mail: [email protected] Background and Aims: Genetic variations in the SNP rs12979860 near the IFNL3 (IL28B) gene affect the likelihood of achieving an SVR in HCV genotype (GT)-1a infected patients following interferonfree therapy with faldaprevir and deleobuvir. Here we report on the frequency of another SNP (rs8099917) and the combined effect on SVR with rs2979860 in the SOUND-C2 trial. Methods: SOUND-C2 was a randomised, open-label Phase 2b study of faldaprevir + deleobuvir ± ribavirin in 362 treatmentnaïve patients with HCV GT-1. Frequency of the rs12979860 and rs8099917 genotypes was assessed by TaqMan-based allelic discrimination PCR analysis from patients in the ribavirincontaining arms and their association with SVR12 calculated in the virologically evaluable set (n = 267). Results: Forty-eight percent (128/267) of patients had the favourable rs8099917 genotype (TT) and 52% the unfavourable rs8099917 genotypes (non-TT). The difference in SVR rates achieved by patients with favourable and unfavourable genotypes was 25% in GT-1a and 15% in GT-1b-infected patients (table). All patients with rs12979860 CC were also rs8099917 TT. Among GT-1a patients, SVR12 was achieved by 71% and 60% of patients with the favourable rs12979860 genotype (CC) and the favourable rs8099917 genotype (TT), respectively. Conclusions: SVR rates were consistently high in GT-1b patients receiving faldaprevir + deleobuvir + ribavirin irrespective of the IFNL3 SNPs. The effect of rs8099917 on SVR was more pronounced


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POSTERS among GT-1a than GT-1b-infected patients. Overall, the rs12979860 CC genotype appeared to be a better predictor of virological success than the rs8099917 TT genotype. Combining the SNPs did not increase predictability over rs12979860 alone. Table: SVR12 for rs12979860 and rs8099917 by genotype rs8099917 Favourable (TT)

Unfavourable (non-TT)

(n/N)

GT-1a

GT-1b

GT-1a

Total rs12979860 Favourable (CC) Unfavourable (non-CC)

60% (36/60)

93% (63/68)

35% (19/54)

GT-1b 78% (66/85)

71% (22/31) 48% (14/29)

97% (33/34) 88% (30/34)

0% (0/0) 35% (19/54)

0% (0/0) 78% (66/85)

P727 VALIDATION AND COMPARISON OF SIX NON-INVASIVE SCORES FOR THE DIAGNOSIS OF LIVER FIBROSIS IN CHILDREN WITH CHRONIC HEPATITIS C S. Barakat1 , W. El-Gendy2 , H. Abou El-Naga1 , M. Salem3 . 1 Pediatrics, Alexandria University, Faculty of Medicine, 2 Clinical Pathology, Alexandria University, 3 Pathology, Alexandria University, Faculty of Medicine, Alexandria, Egypt E-mail: [email protected] Aims: To asses the diagnostic performance of 6 non-invasive fibrosis scores, not validated before in children with chronic HCV infection. Methods: Fibrometer, Hepascore, Forns score, Fibroindex, FIB-4 and APRI were assessed in fifty-two children with HCV, using liver histology as reference standard. The primary end-point was the area under characteristic curves (AUROC) for the detection of significant fibrosis by METAVIR classification. Results: 13% of studied children had significant fibrosis. Hepascore has poor performance (AUROC 0.638), the remaining studied scores have good diagnostic accuracies (AUROCs ranged from 0.921 for APRI to 0.825 for fibroindex). ALL cores showed excellent negative predictive value (NPV >95%) to exclude significant fibrosis, while none of the scores present with a positive predictive value (PPV) good enough (>90%) to reliably predict significant fibrosis in HCV studied children. In multiple stepwise logistic regression including the five scores, APRI and fibrometer (AUROCs 0.921 and 0.857 respectively) were the only scores independently associated to significant fibrosis. Our results strongly suggest that exclusion of significant fibrosis can be made with more than 95% certainty when APRI is lower than 0.68 or fibrometer is lower than 2.94. By contrast, significant fibrosis cannot be predicted with the same certainty. Conclusions: We suggest to perform APRI in all children; those classified by APRI as non significant fibrosis did not need further evaluation due to high NPV. However, children showing significant fibrosis by APRI had to undergo liver biopsy. With this algorithm we obtained 93% accuracy in exclusion of significant fibrosis and saved around 80% of liver biopsy. P728 HEPATITIS C VIRUS (HCV) CORE ANTIGEN ASSAY FOR DETECTION OF ACTIVE HCV INFECTION AMONG HIV-INFECTED MEN WHO HAVE SEX WITH MEN ATTENDING AN STI CLINIC J.W. Vanhommerig1,2 , T.J.W. van de Laar3 , M.S. van Rooijen1 , H.J. de Vries1,4 , A.G.C.L. Speksnijder1 , M. Prins1,4 , S.M. Bruisten1 . 1 Cluster of Infectious Diseases, Department of Research, Public Health Service of Amsterdam, 2 Section of Clinical Virology, Department of Medical Microbiology, Academic Medical Center, 3 Department of Blood-Borne Infections, Sanquin Blood Supply Foundation, 4 Department of Internal Medicine, Center of Infectious Diseases and Immunology Amsterdam (CINIMA), Academic Medical Center, Amsterdam, Netherlands E-mail: [email protected] Background and Aims: Formation of HCV-antibodies may be delayed up to 1 year after infection, especially when one S312

is co-infected with HIV. For detection of early HCV infection and reinfection, commercial HCV-RNA tests are available. However, these tests are time-consuming and expensive, and are therefore currently not suitable for population-wide screening. A commercially available test that may supplement current screening methods, targets the HCV core protein. Methods: During five waves of anonymous surveys at the Amsterdam STI clinic in 2009–2012, all HIV-infected MSM were tested for HCV-antibodies (AxSYM HCV 3.0, Abbott), and HCV-RNA (TMA, Siemens). To evaluate screening with the Architect HCV antigen (HCV-Ag) assay (Abbott), all HCV-RNA positive samples were selected and for each positive sample, two HCV-RNA negative samples were selected as controls. Results: Among 439 HIV-infected MSM, 31 (7.1%) tested positive for HCV-RNA. The HCV-Ag assay showed concordant positive results in 31/31 (100%). A large proportion of MSM who had active HCV infection, i.e., 6/31 (19.4%), were HCV-seronegative at the time of screening and were presumed to have an acute infection. Concordant HCV-RNA-negative results were obtained in 60/62 (96.8%) control-samples; 2/62 (3.2%) gave erroneous results in the HCV-Ag assay and were considered discordant results. Specificity of the HCV-Ag assay was 97.8% (95% CI 92.4%-99.7%). Conclusions: The HCV-Ag assay proved a valuable screening tool for detection of active HCV infection in HIV-infected MSM. Detection of early HCV infections is preferred not only to prevent transmission; treatment success rates are significantly improved when treatment is started earlier after infection. P729 HCV GENOTYPE 1A AND 1B: SIMILARITIES AND DIFFERENCES IN CLINICAL FEATURES, THERAPEUTIC OUTCOME AND PREDICTORS OF RESPONSE F. Morisco1 , V. Di Marco2 , R. Granata1 , S. Camera1 , A. Ippolito3 , M. Margaglione4 , G. Fattovich5 , A. Smedile6 , E. Clery1 , C. Ferraiuoli1 , M. Guarino1 , L. Donnarumma1 , M.R. Valvano2 , M. Milella7 , M.M. Felder8 , G.B. Gaeta9 , P. Gatti10 , P. Tundo11 , M. Barone12 , R. Cozzolongo13 , M. Angelico14 , G. Mazzella15 , T. Santantonio16 , N. Caporaso1 , A. Andriulli3 . 1 Department of Clinical Medicine and Surgery, Gastroenterology Unit, University of Naples ‘Federico II’, Naples, 2 Division of Gastroenterology, University of Palermo, Palermo, 3 Division of Gastroenterology, Casa Sollievo Sofferenza Hospital, IRCCS, San Giovanni Rotondo, 4 Department of Clinical and Experimental Medicine, University of Foggia, Foggia, 5 Division of Gastroenterology, University of Verona, Verona, 6 Department of Medical Science, University of Torino, Torino, 7 Clinic of Infectious Diseases, University of Bari, Bari, 8 Division of Gastroenterology, Central Hospital, Bolzano, 9 Clinic of Infectious Diseases, University of Naples, Naples, 10 DIMO Medical Oncology, University of Bari, Bari, 11 Division of Infectious Diseases, S. Caterina Novella Hospital, Galatina, 12 Division of Gastroenterology, University of Bari, Bari, 13 Division of Gastroenterology, S. de Bellis Hospital, IRCCS, Castellana Grotte, 14 Division of Gastroenterology, University of Tor Vergata, Roma, 15 Division of Gastroenterology, University of Bologna, Bologna, 16 Clinic of Infectious Diseases, University of Foggia, Foggia, Italy E-mail: [email protected] Background and Aims: Clinical trials assessed treatment response rate among HCV-1a and -1b genotype patients as a single group, and it has been unable to uncover differences in treatment outcome between the two subtypes. This study evaluates the baseline similarities and differences between the two HCV-1 subtypes on pre-treatment and on-treatment predictors of response to peginterferon (PEG-IFN) and ribavirin. Methods: 1184 naïve patients with HCV genotype-1 infection were treated with PEG-IFN and ribavirin (1000–1200 mg/day). Between January 2005 and December 2010 15 Italian centers, selected by voluntary participation, took part in the study. The study included


POSTERS 155 (13%) patients infected with subtype 1a and 1029 (87%)with subtype 1b. Results: Six factors differentiate genotype 1a vs genotype 1b: more frequent male sex (74% vs 56%; p < 0.001); age younger than 50 years (63% vs 30%; p < 0.001); value of BMI (24 vs 25; p < 0.042); higher value of platelet count (209×103 vs 186×103 ; p < 0.001); less frequent severe liver fibrosis (70% vs 52%; p < 0.001) and finally less frequent type 2 diabetes comorbidity (5% vs 12%; p < 0.005). All other parameters resulted equally distributed between the two HCV 1 subtypes. Three factors were independently associated in subtype 1a: female gender, IL28B polymorphism, and RVR; and three factors independently associated in subtype 1b: low baseline serum HCV-RNA concentration (400,000 IU/mL (PegIFN/RBV 43.2%, BOC/PegIFN/RBV 71.1%, TVR/PegIFN/RBV 74.1%). SVR rates were 42.6% for PegIFN/RBV, 53.3% for BOC/PegIFN/RBV and 59.6% for TVR/PegIFN/RBV. AE/SAE rates were 81.4%/7.1% for PegIFN/RBV, 91.3%/13.6% for BOC/PegIFN/RBV and 91.4%/11.3% for

TVR/PegIFN/RBV. SVR rates after the treatment of AEs and comedication prescribed for AE treatment are listed in the table. Conclusions: Triple therapy leads to significantly more AEs than PegIFN/RBV. Active management of AEs with co-medication increased SVR rates. Psycholeptics and analgesics were most frequently prescribed during dual and triple therapy.

Treatment-related AEs PegIFN RBV BOC TVR Co-morbidities Prescribed co-medication (%) N05/N06 N02 D07 A02 A03 C09 M01 Most frequent AEs (%) Fatigue Skin Disorder/Rash Nausea Pruritus Anaemia Headache Arthralgia Dysgeusia

PegIFN/RBV

BOC/PegIFN/RBV

TVR/PegIFN/RBV

(%)

SVR (%)

(%)

SVR (%)

(%)

SVR (%)

37.2 20.8 − − 33.9

54.4 57.9 − − 43.5

46.0 32.8 23.0 − 44.9

56.8 54.3 48.5 − 53.5

46.3 38.0 − 42.8 40.2

62.0 65.6 − 63.3 58.9

28.4 18.6 7.7 13.7 4.4 7.1 11.5

36.6 25.8 12.2 14.6 10.8 12.5 9.1

31.5 24.7 21.3 18.0 14.0 11.9 9.9

72.5 31.5 24.8 12.1 14.8 28.9 18.8 2.7

72.5 40.5 29.8 23.7 22.1 30.9 22.9 27.5

68.0 50.5 30.0 33.2 27.3 18.5 16.6 10.2

Co-medication coded to WHO ATC level 2: N05/N06 = psycholeptics/psychoanaleptics; N02 = analgesics; D07 = corticosteroids, dermatological preparations; A02 = antacids, drugs for treatment of peptic ulcerand flatulence.; A03 = antispastic, anticholinergic agents and propulsives; C09 = agents acting on the reninangiotensin system, anti-inflammatory and anti-rheumatic products.

P731 CANNABINOID RECEPTOR 2-63 QQ VARIANT IS ASSOCIATED WITH PERSISTENTLY NORMAL AMINOTRANSFERASE SERUM LEVELS IN CHRONIC HEPATITIS C N. Coppola, R. Zampino, C. Sagnelli, G. Bellini, A. Marrone, M. Stanzione, N. Capoluongo, A. Boemio, C. Minichini, L.E. Adinolfi, S. Maione, E. Miraglia Del Giudice, E. Sagnelli, F. Rossi. Second University of Naples, Naples, Italy E-mail: [email protected] Background and Aims: To evaluate in anti-HCV-positive patients the clinical impact of the rs35761398 single nucleotide polymorphism (SNP) of the CNR2 gene leading to the substitution of Arg (R) of codon 63 of the cannabinoid receptor 2 (CB2) with Gln (Q). Methods: 253 consecutive anti-HCV-/HCV-RNA-positive patients were enrolled: males 53%; median age 52 years (range 18–80), mean serum HCV-RNA 2.6×106 IU/ml (SD: ±5.6×106 ), 63.6% with genotype 1, 73.5% with liver biopsy (Histological Activity Index: 5.93±3.5; fibrosis score according Ishak score 2.3±1.42; steatosis score: 1.19±1.2). Of these 253 patients, 53 were HCV carriers with persistently normal ALT (PNALT group) and 200 had a history of steadily abnormal serum ALT values (abnormal ALT group). All patients were naive for antiviral therapy and were screened for CNR2 rs357661398 SNP by a real-time assay. Results: Subjects in the PNALT group, compared with those in the abnormal ALT group were older (58.5±12 vs 50.7±12.4 years, p = 0.001), more frequently female (66% vs. 42%, p = 0.003), with lower body mass index (24.5±3.1 vs. 26.6±4.6, p = 0.003), and more frequently with HCV genotype 2 (43.1% vs 17.7%, p = 0.0002) and CB2-63 QQ variant (34% vs. 11%, p = 0.0001). Considering all 253 patients, no difference in the demographic, biochemical, or virological data was observed between patients in the different CB2-63 variants. The logistic regression analysis identified CB2-63 QQ, HCV genotype 2, older age and lower BMI as independent predictors of PNALT (p < 0.00001). Conclusions: The CB2–63 QQ variant in HCV patients was independently associated to the PNALT status.


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POSTERS P732 PREDICTIVE VALUE OF THE INTERFERON-LAMBDA 4 GENE (IFN-L4) FOR DIRECT-/ANTIVIRAL TREATMENT IN HEPATITIS C VIRUS INFECTED PATIENTS WITH GENOTYPES 1, 2, 3, AND 4 S. Susser1 , C.M. Lange1 , N. Hamdi2 , T. Berg3 , D. Perner1 , S. Zeuzem1 , C. Sarrazin1 . 1 Medical Clinic 1, Goethe-University Hospital, Frankfurt/Main, Germany; 2 Department of Pharmacology ur and Toxicology, German University Cairo, Cairo, Egypt; 3 Klinik f¨ Gastroenterologie, Sektion Hepatologie, Universit¨ atsklinik Leipzig, Leipzig, Germany E-mail: [email protected] Background and Aims: The TT/DG-polymorphism upstream of IL28B was identified as first potential functional variant for induction of IL28B expression. The DG-allele was described as responsible for expression of IFN-L4, which is associated with impaired clearance of HCV. The power for prediction of treatment outcome of IFN-L4 in HCV genotype 1, 2/3, and 4 infected patients, treated with telaprevir (TPR) and/or (pegylated) interferon-alfa (PEG-IFNa)/ribavirin is largely unknown. Methods: The IFN-L4 host-genotype (gt) (ss469415590) and IL28B/IFN-L3-genotypes (rs12979860/rs8099917) were analyzed in patients with chronic HCV gt1 (n = 385), 2/3 (n = 267), and 4 (n = 197) infections for associations with sustained virologic response (SVR) to antiviral therapy consisting of either (PEG-)IFNa/ribavirin or TPR+PEG-IFNa/ribavirin (n = 79). As controls, healthy people from Germany/Egypt (n = 283)/(n = 96) were included. Results: The ss469415590_TT/DG genotype is associated with SVR in HCV gt1 dual- but not triple-therapy and in gt2/3, but not gt4 infected patients (p < 0.001; p = 0.775; p = 0.020; p = 0.157). In accordance with the findings for rs12979860_CC, ss469415590_TT is associated with higher baseline viral load in gt1 dual and gt2/3 patients (p < 0.001; p < 0.001). In our HCV gt1 infected patients treated with TPR+PEG-IFNa/ribavirin, response is neither predictable through ss469415590, nor by rs12979860 (p = 0.775/0.287). Conclusions: Prediction of response after dual-therapy in HCV gt1 and 2/3 patients is equally possible with genotyping of rs12979860 or ss469415590. In HCV gt4 patients only IFN-L3 but not IFN-L4 is associated with treatment outcome. No treatment association was found for our “real-life” triple therapy cohort, which mainly consisted of former relapsers/non-responders. In summary, despite functional context, IFN-L4 treatment response prediction was not superior to IL28B. P733 NEW HEPATITIS C INFECTION AND RE-INFECTION AMONG HIV CO-INFECTED MEN IN MELBOURNE, AUSTRALIA C. El-Hayek1 , J.S. Doyle1,2,3 , M. Cuevas4 , M. Lim1 , C.K. Fairley4,5 , D.E. Leslie6 , N. Roth7 , B.K. Tee8 , M. Stoove1 , M.E. Hellard1,2,3 . 1 Centre for Population Health, Burnet Institute, 2 Department of Infectious Diseases, The Alfred and Monash University, 3 Department of Epidemiology and Preventative Medicine, Monash University, 4 Melbourne School of Population Health, University of Melbourne, 5 Melbourne Sexual Health Centre, 6 Victorian Infectious Diseases Reference Laboratory, Melbourne, 7 Prahran Market Clinic, Prahran, 8 Centre Clinic, Victorian AIDS Council, Melbourne, VIC, Australia E-mail: [email protected] Background and Aims: Sexual transmission of HCV among HIV infected men-who-have-sex-with-men (MSM) appear to be driving increased HCV notifications in many settings. High re-infection rates after viral clearance also pose prevention and treatment challenges. To determine the magnitude of this problem, we calculate the frequency of new HCV infection and re-infection among frequently testing HIV-infected MSM. Methods: A retrospective cohort analysis was conducted using laboratory HCV test records of all HIV-infected men who S314

attended three high MSM-caseload clinics in Melbourne for regular management (April 2006-December 2011). We defined incident HCV as HCV-antibody seroconversion or newly detected HCV RNA when previously HCV-antibody positive/RNA negative, and HCV re-infection as RNA positive following two negative RNA tests >30 days apart. Results: Among 2190 HIV-infected MSM tested for HCV, 250 tested HCV positive. The number of HIV-infected men tested for HCV increased each year although the testing frequency remained 1.4 tests/person/year. 63 incident infections and 4 re-infections were observed. The prevalence of HCV-infection declined from 12% in 2006 to 7% in 2011 (p = 0.01) with no significant change in HCV incidence rate of 1.55/100PY (95% CI: 1.18–2.04) over the study period. The incidence of re-infection was 4.63/100PY (95% CI:1.74– 12.33). Conclusions: Incident HCV infection among HIV-infected MSM is low and remains stable among this cohort over time. Our results suggest that increased HCV testing is contributing in part to the increase in HCV case notifications among HIV-infected MSM. However, re-infection rates are substantially higher than primary infection rates, suggesting the need for enhanced health promotion messages and testing among this high-risk group. P734 A SYSTEMATIC REVIEW OF INTERVENTIONS TO REDUCE ALCOHOL CONSUMPTION AMONG INDIVIDUALS WITH CHRONIC HCV INFECTION J.S. Doyle1,2,3 , D. Hunt1 , E.J. Aspinall4,5 , S.J. Hutchinson4,5 , D.J. Goldberg5 , T. Nguyen6 , Y. Falck-Ytter7 , R.L. Morgan8 , B. Smith8 , M. Stoove1 , S. Lutchers9 , A.J. Thompson10,11 , S.Z. Wiktor6 , M.E. Hellard1,2,3 . 1 Centre for Population Health, Burnet Institute, 2 Department of Infectious Diseases, The Alfred and Monash University, 3 Department of Epidemiology and Preventative Medicine, Monash University, Melbourne, VIC, Australia; 4 School of Health and Life Sciences, Glasgow Caledonian University, 5 Health Protection Scotland, Glasgow, United Kingdom; 6 Global Hepatitis Programme, World Health Organisation, Geneva, Switzerland; 7 Case and VA Medical Centre, Case Western Reserve University, Cleveland, OH, 8 Centre for Disease Control and Prevention, Atlanta, GA, United States; 9 Centre for International Health, Burnet Institute, 10 Department of Gastroenterology, St Vincent’s Hospital, 11 Victorian Infectious Diseases Reference Laboratory, Melbourne, VIC, Australia E-mail: [email protected] Background and Aims: Alcohol consumption has been shown to accelerate the progression of liver disease among people with chronic HCV infection. The impact of behavioural interventions to reduce consumption of alcohol among people with HCV remains uncertain. This review aims to determine the effectiveness of behavioural interventions to reduce alcohol consumption and liver disease complications, while improving treatment response and quality of life. Methods: A systematic review was conducted following Cochrane guidelines to identify studies of HCV-infected individuals receiving any behavioural interventions to reduce alcohol consumption compared to no intervention published up to 1 May 2013. Interventions included brief alcohol counselling, motivational enhancement therapy, and integrated alcohol education during HCV therapy. Outcomes of interest included alcohol intake; sustained virological response; liver fibrosis, decompensation, or cancer; and quality of life. The review protocol was registered prospectively. Results: Of 1109 unique citations, five studies (two randomised control trials (RCT), three cohorts) met the selection criteria. Alcohol intake was the only outcome reported; no studies reported on mortality, morbidity or quality of life. Measurement of alcohol consumption varied across studies, precluding any quantitative synthesis. One RCT (n = 135) demonstrated a reduction of 4.2


POSTERS drinking days per month; another (n = 334) showed no change in AUDIT score (OR 0.99, 95% CI: 0.96–1.01) at six months. Achieving abstinence at six months was achieved in one (n = 53) cohort (OR 3.36, 95% CI: 2.62–4.10). Conclusions: Alcohol reduction interventions for chronic HCV patients could lead to abstinence or reduce the number of drinking days per month. Practitioners should be encouraged and trained to provide alcohol-reduction counselling. P735 ASSOCIATION OF GENETIC POLYMORPHISMS WITH FIBROSIS PROGRESSION IN CHRONIC HEPATITIS C AFTER FAILURE OF INTERFERON THERAPY N. Tamaki, M. Kurosaki, Y. Yasui, S. Suzuki, T. Hosokawa, K. Tsuchiya, H. Nakanishi, J. Itakura, N. Izumi. Department of Gastroenterology and Hepatology, Musashino Red Cross Hospital, Tokyo, Japan E-mail: [email protected] Background and Aims: Genome-wide association study identified several single nucleotide polymorphisms (SNPs) that are associated with liver fibrosis progression in natural course of chronic hepatitis C. The aim of the present study was to evaluate the role of these SNPs on fibrosis progression after failure of interferon therapy. Methods: A paired liver biopsy was performed in 157 chronic hepatitis C patients who failed to achieve sustained virological response by interferon. Mean period between two biopsies were 5.8 years. Fibrosis progression per year was calculated as difference of METAVIR fibrosis stage between two biopsies. Association between fibrosis progression and genotype of MERTK (rs4374383), TULP1 (rs9380516), GLT8D2 (rs2629751), and IL28B (rs8099917) was analyzed. Results: Fibrosis stage before interferon therapy was F1 in 53%, F2 in 29%, and F3 in 18%. After interferon therapy, fibrosis stage progressed in 62 patients and regressed in 10. The median rate of fibrosis progression was 0.08±0.17/year. Increased rate of fibrosis progression was significantly associated with IL28B (TT 0.03±0.15 vs. TG/GG 0.16±0.18, p < 0.001) and TULP1 (CC 0.06±0.17 vs. CT/TT 0.14±0.18, p = 0.02) but not with MERTK or GLT8D2 genotype. By multivariate analysis, IL28B TG/GG genotype (HR: 1.8, 95% CI: 1.1–3.1, p = 0.01) and older age (HR: 2.0, 95% CI: 1.2–3.4, p = 0.008) was independent predictor of fibrosis progression. Conclusions: The host factors of older age and IL28B TG/GG genotype have strong association with fibrosis progression after failure of interferon therapy. P736 SCREENING FOR HEPATITIS C AND COMPARISON OF DISEASE CHARACTERISTICS AND RESPONSE TO TREATMENT WITH NON-SCREENED PATIENTS M. Sharma1 , S.A. Kaabi1 , N.A. Dweik1 , A.K. John1 , H. Wani1 , M.A. Mohannadi1 , M.F. Derbala1 , R. Yacoub1 , R.B. Thandassary1 , F. Pasic1 , M.E. Bedawi2 , M.T. Butt1 , R. Singh1 . 1 Hamad Medical Corporation, 2 Weill Cornell Medical College in Qatar, Doha, Qatar E-mail: [email protected] Background and Aims: Screening for Hepatitis C has been found to be beneficial in high-risk individuals and baby boomers. To screen for Hepatitis C in average and high risk individuals in Qatar. Secondary aims included comparing the disease characteristics and response to treatment among screened and nonscreened hepatitis C patients. Methods: A community-based survey was done to screen for Hepatitis C antibodies in the average and high-risk population over a 16 months period in the capital city of Doha. Screening for antibodies was done using a rapid immunochromatographic assay. All patients detected to have at least stage 1 fibrosis on the

liver biopsy (Scheuer classification) were treated with pegylated interferon and ribavirin. Results: 13704 people were screened and 272 (2%) had positive antibodies to hepatitis C. During the same period, 237 non-screened patients with hepatitis C were referred for treatment. Screened Group (SG) and non-screened groups (NSG) had similar baseline characteristics. ALT, AST and fibrosis on the liver biopsy were significantly lower in the SG as compared to NSG (p = 0.04, 0.04 and 0.01 respectively). The response to treatment was not statistically significant in the SG as compared to NSG (SVR 61.7% versus 69.1% p = 0.55). Among the SG, average risk patients had significantly lower ALT levels (p = 0.04) and similar response to treatment as the high-risk individuals (SVR 63.2% versus 61% p = 0.87). Conclusions: Screening detects Hepatitis C at an early stage but does not result in better response to the treatment. Average and high-risk screened patients have a similar response to antiviral treatment. P737 DEVELOPMENT OF A MODEL TO PREDICT PROGRESSION TO CIRRHOSIS IN CHRONIC HEPATITIS C PATIENTS WHO FAILED ANTIVIRAL THERAPY 1 S. Lens1 , F. Torres2 , I. Garc´ıa-Juarez ´ , M.C. Londono ˜ 1 , S. Mart´ınez1 , ´ R. Miquel3 , A. Garc´ıa-Criado4 , R. Gilabert4 , C. Bru4 , J.M. SanchezTapias1 , X. Forns1 . 1 Liver Unit, Hospital Cl´ınic, IDIBAPS, CIBEREHD, 2 Bioestatistics Platform and Data Management Unit, University of Barcelona, Hospital Cl´ınic, 3 Pathology Department, 4 Radiology Department, Hospital Cl´ınic, IDIBAPS, CIBEREHD, Barcelona, Spain E-mail: [email protected] Background and Aims: Progression of liver disease may occur in non-cirrhotic patients with chronic HCV infection who fail antiviral therapy. The aim of this study was to generate a model among nonresponders patients to identify those at higher risk of developing liver cirrhosis and decompensation. Methods: Non-cirrhotic patients seen at our Unit between April 1990 and May 2003 who failed antiviral therapy were included. All patients were followed prospectively with periodic clinical, analytical and ultrasonographic examinations. A comparative analysis was performed between patients who developed cirrhosis and those who did not. Results: 896 non cirrhotic patients received treatment during that period. Baseline liver biopsy showed F0–F1 in 76%. 495 patients (55%) failed to achieve sustained virological response. After a mean follow-up of 12 years, 126 (25%) patients developed cirrhosis, 26 had clinical decompensation and 26 hepatocellular carcinoma. Using a Cox regression model, baseline variables independently related to cirrhosis development were: F ≥ 2 (HR 2.61), age >40 years (HR 2.01), GGT >40 UI/L (HR 2.35) and AST >100 UI/L (HR 1.76). Models including these variables (with or without liver biopsy) could predict cirrhosis development at 3, 5, 7 and 10 years with AUC values between 0.83 and 0.89. APRI, FIB-4 and Forns score at baseline also had a high predictive value, with AUC between 0.76 and 0.89 at 3, 5, 7 and 10 years. Conclusions: In non responder patients, simple baseline variables (including APRI, FIB-4 or Forns scores) can predict cirrhosis development. These models could be useful to identify patients requiring treatment more urgently.


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POSTERS P738 NO CLINICALLY RELEVANT DRUG–DRUG INTERACTIONS BETWEEN FALDAPREVIR AND PEGYLATED INTERFERON a-2a PLUS RIBAVIRIN IN HCV-INFECTED PATIENTS: PHARMACOKINETIC ANALYSES FROM TWO PHASE II STUDIES M. Bourliere ` 1 , M. Sulkowski2 , M.P. Manns3 , T. Asselah4 , P. Ferenci5 , S. Pol6 , T. Berg7 , J. Lalezari8 , A.-M. Quinson9 , Y. Datsenko10 , opital Saint Joseph, Marseille, J. Scherer9 , C.-L. Yong9 , F. Huang9 . 1 Hˆ France; 2 Johns Hopkins University School of Medicine, Baltimore, MD, United States; 3 Hannover Medical School, Hannover, Germany; 4 Hˆ opital Beaujon, APHP, University Paris-Diderot and INSERM CRB 3, opital Clichy, France; 5 Medical University of Vienna, Vienna, Austria; 6 Hˆ atsklinikum Leipzig A¨ oR, Leipzig, Cochin, Paris, France; 7 Universit¨ Germany; 8 Quest Clinical Research and University of California-San Francisco, San Francisco, CA, 9 Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT, United States; 10 Boehringer Ingelheim Pharma GmbH & Co KG, Biberach, Germany E-mail: [email protected] Background and Aims: Faldaprevir is a potent HCV NS3/4A protease inhibitor in development for the treatment of chronic HCV infection in combination with ribavirin and pegylated interferon a-2a (PegIFN), and in interferon-free combinations. Here we report the pharmacokinetic (PK) interactions between faldaprevir and ribavirin and PegIFN. Methods: PK interactions were measured in two Phase Ib-II studies. In study 1, treatment-naïve patients (N = 26) received 14 days (D) faldaprevir followed by 14D faldaprevir plus PegIFN/ribavirin. The effect of PegIFN/ribavirin on steady-state (SS) faldaprevir was assessed by comparing PK profiles on D28 with D14. In study 2, the effect of 120 and 240 mg faldaprevir on ribavirin PK was assessed versus placebo in a substudy (N = 49) at week (W) 10 of 24W treatment; the effect on PegIFN was evaluated by comparison of faldaprevir trough values (PegIFN/ribavirin + faldaprevir vs PegIFN/ribavirin + placebo) at W12 and W24. Results: The geometric mean ratio (GMR) (90%CI) of faldaprevir plus PegIFN/ribavirin versus faldaprevir alone was 107.4 (96.9– 119.1), 110.5 (96.5–126.4) and 106.8 (94.5–120.6) for AUCt,ss , Cmax,ss and Cmin,ss , respectively. The GMR of ribavirin/faldaprevir versus ribavirin/placebo for ribavirin AUCt,ss was 0.81 and 0.82 for 1000 mg and 1200 mg ribavirin, respectively (for the faldaprevir 120 mg regimen), and 1.06 and 0.83, respectively (for the faldaprevir 240 mg regimen). The trough PegIFN concentration GMR was within ±10% for faldaprevir vs placebo at W12 and W24. Conclusions: These data indicate that faldaprevir has no clinically relevant effect on exposure of ribavirin or PegIFN, and PegIFN or ribavirin have no effect on the PK of faldaprevir. P739 RISK FACTORS AND TREATMENT OUTCOMES IN GENOTYPE 1 AND GENOTYPE 4 HEPATITIS C INFECTED PATIENTS FROM A REAL WORLD DATA ANALYSIS IN EUROPE C. Moreno1 , Y. Horsmans2 , R. Flisiak3 , H. Van Vlierberghe4 , E. Trepo1 , P. Starkel2 , J. Jaroszewicz3 , A. Geerts4 , M. Pisini5 , L. Lantican6 , D. Koletzki6 . 1 Erasme University Hospital, 2 Saint Luc University Hospital, Brussels, Belgium; 3 Medical University of Bialystok, Bialystok, Poland; 4 University Hospital Ghent, Ghent, 5 Janssen Pharmaceutica NV, 6 Janssen Diagnostics BVBA, Beerse, Belgium E-mail: [email protected] Background and Aims: Despite improvement in treatment of chronic hepatitis C (CHC), there seems to be an unmet need for patients infected with other genotypes (G) than G1 in real world settings. This was assessed by comparing clinical outcomes of patients with G1 and G4. Methods: Routine clinical practice data were collected from Belgium and Poland using a web-based registry. Univariate statistical data analysis was performed. S316

Results: Demographics: N = 293 G1 patients mainly consisting of Caucasians (91%) and n = 75 G4 patients mainly consisting of BlackAfricans (54%) and Caucasians (42%) were studied. Nosocomial virus transmission was equally observed in both genotypes (57%). Injecting-drug-use-related transmission was higher in G1 (22%) than G4 (13%), whereas sexual transmission was lower in G1 (5%) than G4 (13%). HIV co-infection was observed in 6% of G1 versus 15% of G4 patients. Risk factors: A higher percentage of patients with baseline BMI >25 was seen in G4 (53%) and differences were observed for comorbidities with higher frequencies for hypertension, diabetes and hypercholesterolemia and higher prescription frequencies of certain concomitant medication potentially interacting with CHC treatment in G4 compared to G1 (table). Treatment and treatment response: PegIFN+RBV-based regimen contributed with 76% to treatments. In terms of ultimate treatment response per patient, the G4 cohort showed a higher percentage of treatment failure (65%) than the G1 cohort (52%). Table: Potential risk factors for lower treatment success Observed risk factors a

BMI >25 HIV Co-infection Having ≥1 concurrent illness b Leading concurrent illnesses b Hypertension Diabetes Hypercholesterolemia Renal failure Leading concomitant medication b Antihypertensives Calcium channel blockers HIV-Antivirals Beta blockers Statins a b

G1 (n = 293)

G4 (n = 75)

42% 6% 46%

53% 15% 60%

18% 7% 3% 1%

32% 17% 5% 5%

20% 5% 7% 6% 3%

30% 18% 17% 12% 3%

Total number of records with available information (G1 n = 158; G4 n = 45). Total number of treated patients (G1 n = 234; G4 n = 60).

Conclusions: The data suggest that G4 patients had a higher prevalence of risk factors with potential negative impact on treatment response, thus highlighting the need to identify disease management strategies to improve treatment outcomes. P740 PHARMACOKINETICS OF FALDAPREVIR IN HEALTHY SUBJECTS FOLLOWING CO-ADMINISTRATION WITH ITRACONAZOLE K. Marzin1 , R. Koenen1 , N. Strelkowa1 , K.-P. Kammerer1 , M. Elgadi2 , F. Huang3 . 1 Boehringer Ingelheim Pharma GmbH & Co. KG, Ingelheim, Germany; 2 Boehringer Ingelheim Canada Ltd/Ltée., Burlington, ON, Canada; 3 Boehringer Ingelheim Pharmaceuticals, Inc., Ridgefield, CT, United States E-mail: [email protected] Background and Aims: Faldaprevir is a potent inhibitor of HCV NS3/4A protease in Phase III development for the treatment of chronic HCV infection. Since faldaprevir is a CYP3A4/Pglycoprotein substrate, we investigated the extent to which the pharmacokinetics of faldaprevir is influenced by co-administration of itraconazole, a strong CYP3A4/P-glycoprotein inhibitor. Methods: This was an open-label, fixed-sequence study with intrasubject comparison to evaluate the effect of itraconazole 200 mg QD on the pharmacokinetics of faldaprevir. Faldaprevir 120 mg BID was administered on Day 1 and then 120 mg QD from Day 2 until end of study (total 10 days); after 6 days of faldaprevir, itraconazole 200 mg QD (except 200 mg BID on first day) was added to faldaprevir for an additional 4 days. Intensive PK sampling was performed after 6 days of faldaprevir treatment and after 4 days of faldaprevir + itraconazole treatment.


POSTERS Table (abstract P740): Faldaprevir PK parameters: adjusted geometric mean ratios Pharmacokinetic parameter

AUCt,ss [ng·h/mL] Cmax,ss [ng/mL]

Adjusted gMean

Intra-individual gCV, %

Faldaprevir + itraconazole (test) (N = 17)

Faldaprevir alone (reference) (N = 17)

Ratio of test to reference treatment, % (LL, UL) (N = 17)

59,454.33 5,027.05

29,944.19 2,783.02

198.55 (182.43, 216.09) 180.63 (165.68, 196.93)

Results: Eighteen subjects received trial medication and 17 completed the trial. The table shows steady-state pharmacokinetic parameters along with geometric mean ratio of faldaprevir with vs. without itraconazole. Conclusions: Co-administration of faldaprevir with itraconazole resulted in an approximately 2-fold increase in faldaprevir steady-state exposure. Administration of faldaprevir alone or in combination with itraconazole was generally safe and well tolerated. No dose adjustment is required when faldaprevir is coadministered with itraconazole. P741 PHASE ANGLE IS ASSOCIATED WITH ADVANCED FIBROSIS IN HCV PATIENTS M.S. Dorna, N.A. Costa, L.F. Campos, F.G. Romeiro, L.Y. Sassaki, S.A.R. Paiva, M.F. Minicucci, G.F. Silva. Internal Medicine, State University of Sao Paulo, Botucatu, Brazil E-mail: [email protected] Background and Aims: Hepatitis C is the major cause of chronic liver disease worldwide. It is also known to silently evolve to liver cirrhosis and hepatocellular carcinoma. For more than 20 years Bioelectrical Impedance has been used to assess nutritional status. More recently, Phase Angle (PhA) derived from bioelectrical impedance has been used as a predictor tool in different clinical situations, including liver diseases. Objective: Our aim was to associate PhA and advanced fibrosis in patients with hepatitis C virus (HCV). Methods: Study conducted in a reference treatment centre in Brazil. Written informed consent was obtained from all patients. Inclusion criteria were the confirmed diagnosis for HCV and a previous liver biopsy for fibrosis staging. Exclusion criteria were ascites and age 100% being considered clinically relevant. Since HI may alter protein binding, GS-5816 free fraction (%) was also determined. Results: All subjects completed the study; no subject discontinued due to an AE. All treatment-emergent AEs were Grade 1 (mild), except for one Grade 2 (moderate, ascites: severe HI subject). Total plasma exposures (AUC) were similar in subjects with moderate or severe HI compared to NF. Cmax was lower in subjects with HI but was not considered clinically impactful as overall exposure to GS-5816 was unaltered. GS-5816 free fraction increased with HI and unbound clearance decreased with HI. Table: GS-5816 PK parameter comparison GS-5816 PK parameter

AUCinf AUClast Cmax

GMR% (90% CI) Moderate HI:NF (N = 10:10)

Severe HI:NF (N = 10:10)

83.0 (57.5, 120) 82.0 (56.8, 118) 59.4 (37.8, 88.7)

114 (74.7, 173) 105 (68.7, 160) 47.2 (29.3, 76.0)

Conclusions: No clinically relevant changes in GS-5816 exposures were observed in subjects with HI compared to those with NF. GS-5816 may be administered without dose adjustment to patients with mild, moderate, or severe hepatic impairment.


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POSTERS P743 IMPACT OF BASELINE SERUM FERRITIN ON SURVIVAL OF PATIENTS WITH CHRONIC HEPATITIS C AFTER A 15 YEARS FOLLOW-UP 1 1 C. Jez ´ equel ´ , E. Bardou-Jacquet1 , Y. Desille ´ , I. Renard1 , M. Latournerie1 , P. Houssel-Debry1 , A. Le Gruyer1 , A. Guillygomarc’h1 , C. Le Lan1 , P. Brissot1 , R. Moirand1 , ´ 2 , B. Turlin3 , D. Guyader1 . 1 Liver Y. Deugnier1 , F. Laine´ 1 , O. Loreal Disease Department, 2 Inserm U 991, 3 Pathology Department, CHU Pontchaillou, Rennes, France E-mail: [email protected] Background and Aims: Increased serum ferritin (SF) and liver iron are commonly observed in chronic viral hepatitis C (CHC). A correlation with severity of fibrosis, reponse to antiviral therapy and incidence of hepatocellular carcinoma (HCC) have been suggested in cross-sectional studies. The real impact is still debated. The aim of this study was to investigate the impact of baseline SF on long-term survival. Methods: Patients were selected from a single-center cohort of 4293 CHC(+) consecutive patients. Survival at the date of 15/09/2013 was obtained from death certificate for 3361 patients. Inclusion criteria were HCV-RNA positivity and availability of SF at baseline. Liver fibrosis was determined using liver biopsy (n = 959) and/or non-invasive tests (elastography of fibrotest® ). Survival was studied according to SF level: group 1 (SF < 100 mg/L); group 2 (SF = 100– 300 mg/L); group 3 (SF > 300 mg/L). A Cox model was built to take into account age, gender, therapy and baseline stage fibrosis. Results: 1451 patients were included (mean age 43.5±13.7 years; male 59.8%; fibrosis n = 1173, F0F1 = 60.2%, F2 = 16.8%, F3 = 10%, F4 = 13%); median SF = 171 mg/L [IQR 80–343]; median follow-up = 14.7 years [9.5–18.7]). The overall survival (261 deaths and 55 HCC) was 83.5% after 15 years of follow-up. The survival was lower in group 3 than in groups 1/2 for patients with F0/F1/F2 (p < 0.0001) but not for patients with F3/F4 fibrosis score (p = 0.42). In multivariate analysis, when fibrosis and age were introduced in Cox model, SF was no longer associated with survival.

P744 DEPDC5 GENE LOCUS ASSOCIATES WITH SEVERE LIVER FIBROSIS IN INDIVIDUALS WITH CHRONIC HCV INFECTION M.A. Burza1 , B.M. Motta1,2 , A.L. Fracanzani3 , A. Aghemo3 , S. Fargion3 , M. Colombo3 , J. Fischer4 , T. Berg4 , L. Valenti3 , S. Romeo1,5 . 1 Department of Molecular and Clinical Medicine, University of Gothenburg, Gothenburg, Sweden; 2 University of Milan, Milan, 3 Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Universit` a degli Studi di Milano, MIlan, Italy; 4 Department of Hepatology, University Hospital Leipzig, Leipzig, Germany; 5 Department of Medical and Surgical Sciences, University Magna Graecia, Catanzaro, Italy E-mail: [email protected] Background and Aims: Two genome-wide association studies identified associations between the DEP domain containing 5 (DEPDC5) and the MHC class I polypeptide-related sequence A (MICA) loci and hepatocellular carcinoma (HCC) in Asian subjects with chronic hepatitis C infection (CHC). Since the genetic architecture varies among individuals from different ethnic groups, the aim of this study was to investigate the effect of DEPDC5 rs1012068 (T>G), and MICA rs2596542 (C>T) variants on HCC and fibrosis risk in Europeans with CHC. Methods: The genetic variants were genotyped in an Italian cohort of CHC individuals with (n = 149) and without HCC (n = 328) from Milan. Among all the individuals (n = 477), n = 300 showed severe fibrosis (F3/4) whereas n = 177 had mild/absent fibrosis (F0/1). Significant associations were validated in an independent cohort from Leipzig with a full spectrum of liver disease (n = 630). Results: Neither the DEPDC5 nor MICA variants were associated with HCC in the Milan cohort. An increased risk of developing severe fibrosis was observed in DEPDC5 G allele carriers from Milan (OR = 1.46; 95% CI: 1.04–2.04; p-value=0.027) as well as in DEPDC5 G allele carriers from Leipzig (OR = 1.57; 95% CI: 1.14–2.16; p-value=0.006) and in the overall series pooling in the two cohorts (OR = 1.32 95%C.I. 1.08–1.61; p-value=0.007), after adjusting for age and gender. Conclusions: We did not replicate the association between DEPDC5 rs1012068 and MICA rs2596542 variants with CHC-related HCC in Europeans. However, the DEPDC5 rs1012068 variant was a risk factor for severe fibrosis in Europeans with CHC and might indirectly favor carcinogenesis. P745 SEQUENCING OF INITIAL AND RECURRENT HCV INFECTIONS IN HIV-INFECTED MEN WHO HAVE SEX WITH MEN (MSM) M. Macartney1 , A. Ricciardi2 , A. Rodger2,3 , D. Webster1 , D. Karageorgopoulos4 , T. Fernandez2 , F. Ferro2 , J. Sasadeusz5 , S. Bhagani2 . 1 Department of Virology, 2 Infectious Diseases and HIV, Royal Free London NHS Foundation Trust, 3 Research Department of Infection, University College London (UCL), 4 Infectious Diseases, Royal Free London NHS Foundation Trust, London, United Kingdom; 5 Infectious Diseases, The Alfred Hospital, Melbourne, VIC, Australia E-mail: [email protected]

Conclusions: The link between SF and mortality, ascertained in univariate analysis for F ≤ 2 fibrosis score disappeared in multivariate model suggesting absence of direct influence of iron in survival of CHC patients.

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Background and Aims: There is on-going controversy regarding recurrent HCV infection versus reactivation following SVR or spontaneous clearance (SC) in HIV positive MSM. We sequenced 3 viral regions to determine if these were de novo infections or recurrence. Methods: Using standard definitions we identified acute HCV cases from 2002 to 2013. Sequences from HCV protease, polymerase and core/env1 regions were aligned using maximum parsimony phylogenetic trees to identify sequence relatedness from initial and subsequent acute hepatitis episodes. Results: 154 cases of acute HCV (79% Genotype 1) were identified in MSM; average age 38 years. 17 (11%) had SC and 69% (70/101) of those treated achieved SVR. 29% (25/87) had a second episode


POSTERS . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .

15

at primary (open squares) or secondary infection (solid squares). A further 5 switched from genotype 1a to 4 or vice versa; only 1 had evidence for possible re-activation of a pre-existing genotype 1a strain following treatment. Conclusions: We have found a high rate of recurrent acute HCV after SC or SVR in HIV positive MSM who are undergoing de novo re-infection with different genotype 1a strains although a few are re-infecting with a different genotype. Only 1 had evidence of possible re-activation of the primary infecting genotype 1 strain.

17 12 6 26

P746 VITAMIN D RELATED PATHWAY GENE POLYMORPHISMS ENHANCED THE PREDICTION OF TREATMENT RESPONSE IN ASIAN CHRONIC HEPATITIS C-INFECTED PATIENTS WITH UNFAVORABLE IL-28B rs12979860 GENOTYPES K. Thanapirom1 , S. Suksawatamnuay1 , W. Sukeepaisarnjaroen2 , P. Tangkijvanich3 , S. Treeprasertsuk1 , P. Thaimai1 , S. Akkarathamrongsin4 , Y. Poovorawan4 , P. Kullavanijaya1 , P. Komolmit1 . 1 Medicine, Chulalongkorn University, Bangkok, 2 Medicine, Srinagarind Hospital, Khon Kaen, 3 Biochemistry, 4 Pediatrics, Chulalongkorn University, Bangkok, Thailand E-mail: [email protected]

31 16 11

10 11 10 31 9 4 22 17 9

22 H77-NS3 . 16 . . 6 . 15 . 4 . . 12 26 Figure 1.

of acute HCV a median of 3.8 years later. All had on-going risk behaviours. Sequences from initial and subsequent HCV infections were available in 19. 13 primary genotype 1 infections had evidence of secondary acute infection with a different genotype 1a strain. Figure 1 shows the phylogenetic tree from NS3 protease sequences

Background and Aims: IL-28B rs12979860 genotype CT/TT is widely accepted predictor of unfavorable treatment outcome in chronic hepatitis C (CHC) infection. However, some patients who have IL-28B rs12979860 genotypes CT/TT achieve treatment response. Recent study found vitamin D serum level and its common genetic polymorphism CYP27B1-1260 rs10877012 was independent predictor of SVR in poor-response IL28B genotypes. We aimed to determine role of using functionally genetic polymorphisms of vitamin D pathway for predicting antiviral response in this poor response group of patients. Methods: 583 Asian patients with CHC infection treated pegylated interferon-alfa based regimen were enrolled. One hundred and seventeen patients (20.4%) had IL-28B rs12979860 genotypes CT/TT. Twelve common genetic variant of vitamin D pathway including CYP27B1-1260 (rs10877012), CYP2R1 (rs2060793,rs12794714), GC (rs4588,rs7041, rs222020,rs2282679), VDR (rs2228570 Fokl, rs1544410 Bsml, rs757343 Msel, rs7975232 Apal and rs731236 Taql) were genotyped. Results: Factor associated with SVR in patients with IL28B rs12979860 genotype CT/TT was shown in Table 1. Multivariate analysis showed VDR rs7975232 Apal TT genotype (OR = 3.76, 95% CI: 1.28–11.04, p = 0.01) and baseline HCV-RNA 75 years old in the Control group (Elderly Control) with 74 patients who developed HCC at 3.25, age, genotype, gender, race, diabetes and other patient characteristics as independent risk factors. Results: 150,958 out of 360,857 unique HCV CCR patients met study inclusion criteria. Patients with FIB-4 >3.25 experienced a 4-fold increase in the risk of death [HR = 4.23 (4.08–4.38)]. Initiating treatment significantly reduced this risk by 25% [H.R = 0.748 (0.70– 0.78)] even if patients initiated treatment after FIB-4 >3.25. Conclusions: A FIB-4 score >3.25 is a strong predictor of mortality risk in patients with HCV. Clinicians should use changes in FIB-4 scores to motivate untreated patients to initiate treatment. P762 TEMPORAL CHANGES IN HEPATITIS C VIRUS GENOTYPE 3A DISTRIBUTION AMONG PEOPLE WHO INJECT DRUGS IN VANCOUVER, CANADA B. Jacka1 , T. Applegate1 , A.F. Poon2 , R. Harrigan2 , G.J. Dore1 , A. Olmstead3 , K. DeBeck2,4 , M.-J. Milloy2,5 , F. Lamoury1 , C. Woods2 , Z. Brumme2,6 , S. Dobrer2 , V. Dias Lima2,7 , J. Montaner2,7 , J. Joy2 , B.D. Marshall8 , T. Kerr2,7 , E. Wood2,7 , M. Krajden3 , J. Grebely1 . 1 The Kirby Institute, Viral Hepatitis Clinical Research Program, University of New South Wales, Sydney, NSW, Australia; 2 British Columbia Centre for Excellence in HIV/AIDS, St Paul’s Hospital, 3 British Columbia Centre for Disease Control, 4 School of Public Policy, Simon Fraser University, 5 Department of Family Practice, University of British Columbia, 6 Faculty of Health Sciences, Simon Fraser University, 7 Department of Medicine, Division of AIDS, University of British Columbia, Vancouver, BC, Canada; 8 Department of Epidemiology, Brown University, Providence, RI, United States E-mail: [email protected] Background and Aims: IFN-free DAA-based therapy has improved the feasibility of HCV treatment as prevention among people who inject drugs (PWID). However, IFN-free DAA-based therapies have reduced efficacy among people with HCV genotype 3a. We characterized trends in HCV genotype 3a infection and associated factors among PWID in Vancouver, Canada. Methods: Data were derived from two ongoing prospective cohorts of PWID (VIDUS and ARYS, 1996–2012). Participants HCV antibody

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POSTERS positive at baseline and those with HCV antibody seroconversion during follow-up were tested for HCV RNA and sequenced (CoreHVR1/NS5B). Year of infection was estimated as one year after selfreported initiation of injecting. Trends in HCV genotype distribution were evaluated. Factors associated with genotype 3a infection were assessed using logistic regression. Results: Among 818 participants, HCV genotype prevalence was: G1a: 52% (n = 422), G1b: 6% (n = 46), G2a: 3% (n = 22), G2b: 6% (n = 53), G3a: 32% (n = 263), G4a: G SNP is associated in subjects with NAFLD to a diagnosis of NASH, and in patients with CHC with the severity of steatosis and fibrosis. We assessed in patients with Gt1 CHC the relation between PNPLA3 SNPs and histological features of NASH, and of PNPLA3 SNP and NASH on the severity of fibrosis. Methods: 254 consecutive patients with Gt1 CHC were studied. PNPLA3 rs738409 C>G SNP was assessed. Biopsies were scored for staging and grading (Scheuer). Steatosis and features of NASH were also assessed (Bedossa). Results: Histological features of NASH were present in 31.5% of patients. PNPLA3 rs738409 C>G (“G” allele) was found in 40.9% (30.3% CG, 10.6% GG). A “G” allele was independently linked with steatosis ≥30% (OR 2.005, 95% CI 1.015–3.961, p = 0.04) and its prevalence was higher in patients with features of NASH (40/80 in PNPLA3 “G” carriers vs 64/174 in PNPLA3 “CC”, p = 0.04), this association being confirmed at multivariate analysis (OR 2.109, 95% CI 1.194–3.726, p = 0.001) after correction for age and HOMA. Patients with ≥F3 fibrosis had a comparable prevalence of PNPLA3 alleles (29/74 in “G” carriers vs 75/180 in CC, p = 0.71), with a higher prevalence of NASH compared to no NASH (39/74 vs 41/180, p < 0.001). This was confirmed after correction for age, severe necroinflammatory activity, and steatosis ≥30% (OR 2.466, 95% CI 1.271–4.782, p = 0.008). Conclusions: The PNPLA3 C>G SNP has a major link to histological features of NASH among Gt1 CHC patients, thus being indirectly to the severity of fibrosis.

Background and Aims: Fibrosis progression in Hepatitis C virus (HCV) infected patients varies greatly between individuals. Since chemokines recruit immune cells to the infected liver and thus S326


POSTERS P769 SINGLE NUCLEOTIDE POLYMORPHISMS NEAR IL28B AND IL28A GENES AND RISK OF LIVER CIRRHOSIS AMONG PATIENTS WITH CHRONIC HEPATITIS C VIRUS INFECTION M.-H. Lee1 , H.-I. Yang2 , T.-W. Shaw2 , S.-N. Lu3 , Y.-J. Lin2 , C.-L. Jen2 , Y. Yuan4 , G. L’Italien4 , C.-J. Chen2 , R.E.V.E.A.L.-HCV Study Group. 1 Natioal Yang-Ming University, 2 Genomics Research Center, Academia Sinica, Taipei, 3 Department of Gastroenterology, Kaohsiung Chang-Gung Memorial Hospital, Kaohsiung, Taiwan; 4 Global Health Economics and Outcome Research, Bristol-Myers Squibb, Princeton, NJ, United States E-mail: [email protected] Background and Aims: The study aimed to evaluate the associations with the development of cirrhosis for the two candidate SNPs (rs8099917 and rs12979860) and other SNPs near IL28A and IL28B genes among patients infected with HCV. Methods: There were 1095 participants seropositive for antibodies against HCV and seronegative for HBsAg in R.E.V.E.A.L-HCV cohort. A total of 72 SNPs near IL28A and IL28B genes were genotyped using Illumina VeraCode GoldenGate genotyping assay. The ascertainment of cirrhosis was by the regular follow-up by ultrasonography and the computerized data linkage with the National Health Insurance database. In addition, the vital status of our study participants was followed by the computerized linkage with the National Death Certification system. All of the participants were followed from 1991 to Dec, 31, 2010. The adjusted hazards ratios (HRadj ) and 95% confidence interval (CI) were estimated by Cox’s regression models. Results: There were 7.9% cirrhosis and 8.5% non-cirrhosis carried TG/GG on rs8099917 (p = 0.84); 9.8% cirrhosis and 10.0% noncirrhosis carried TT or TC on rs12979860 (p = 0.93). There were no additional SNPs near IL28A or IL28B genes associated with cirrhosis. The elevated serum levels of HCV RNA was the strong risk factor for cirrhosis, with the HRadj of 3.28 (1.87–5.75) for low RNA levels and 4.14 (2.42–7.11) for high RNA levels, compared with participants with undetectable RNA (p for trend G and MERTK rs4374383 A>G SNPs were also assessed. Results: MERTK A>G SNP distribution was similar in cases compared to controls (p = 0.99). In the entire cohort, MERTK AA genotype (OR 0.25, 95% CI 0.10–0.58, p = 0.001) was independently associated with severe steatosis together with PNPLA3 GG status (OR 2.18, 95% CI 1.32–3.59, p = 0.002). In the high-risk group of PNPLA3 GG patients, severe steatosis was observed in none patients with MERTK AA (0/11) compared to 39% (33/84) with MERTK GG/GA genotype (p = 0.01). The presence of fibrosis >F1 was independently linked to MERTK AA genotype in Sicilian cohort only (OR 0.28; 95% CI 0.11–0.69, p = 0.006), but not in the Northern Italy and in the entire cohorts. However, when excluding subjects with BMI > 40 kg/m2 from the entire cohort F2–F4 fibrosis was observed in 19.2% patients with MERTK AA compared to 30.3% with MERTK GG/GA (p = 0.04), being this data confirmed at multivariate analysis (OR 0.44; 95% CI 0.21–0.89, p = 0.02). Conclusions: MERTK AA genotype is protective against severe steatosis in patients with NAFLD, especially in those at high risk because of PNPLA3 GG genotype, while its effect on liver fibrosis needs to be further investigate. P817 EARLY MENOPAUSAL STATUS IS ASSOCIATED WITH THE SEVERITY OF LIVER FIBROSIS IN ITALIAN PATIENTS WITH NON-ALCOHOLIC FATTY LIVER DISEASE S. Petta1 , E. Vanni2 , L. Valenti3 , L. Miele4 , M. Bianchini5 , A.L. Fracanzani3 , E. Bugianesi2 , A. Craxì1 , S. Fargion3 , A. Grieco4 , C. Camma` 1 , E. Villa5 . 1 Sezione di Gastroenterologia, DiBiMIS, University of Palermo, Palermo, 2 Department of Medical Sciences, Division of Gastroenterology and Hepatology, University of Torino, Torino, 3 Department of Pathophysiology and Transplantation, Section Internal Medicine, Universit` a degli Studi, Fondazione Ca’ Granda IRCCS Ospedale Maggiore Policlinico, Milan, 4 Institute of Internal Medicine, School of Medicine, Catholic University of the Sacred Heart, Roma, 5 Department of Gastroenterology, University of Modena and Reggio Emilia, Modena, Italy E-mail: [email protected] Background and Aims: Contrasting data have been reported on the effect of gender and menopausal status on the susceptibility to nonalcoholic fatty liver disease (NAFLD) development and liver damage progression in NAFLD. We assessed whether menopausal status is associated with severity of liver fibrosis in NAFLD. Methods: In 244 consecutive females and 244 age-matched males with biopsy-proven NAFLD, from Italian referral centers, we assessed anthropometric, biochemical, and metabolic features, including menopausal status (self-reported); liver biopsy was scored according to Kleiner. Results: In the entire cohort, at multivariate logistic regression adjusted for age, BMI, type 2 diabetes, serum lipids, and presence of NASH, male gender (OR 1.408, CI 0.779–2.542, p = 0.25) respect to women at reproductive age was not associated with fibrosis >F1, while a trend was observed for menopause (OR 1.752, CI 0.956–3.208, p = 0.06). Accordingly, in female patients menopause (OR 3.079, 95% CI 1.136–8.345, p = 0.02) was associated with fibrosis >F1, independently of the aforementioned confounding factors. Interestingly, in age and multiple covariates-adjusted analysis, we observed a strong effect on liver fibrosis of early (OR 3.038, CI 1.083–8.527, p = 0.03) only, but not of late (OR 2.291, CI 0.771–6.803, p = 0.13) menopause. Conclusions: In female patients with NAFLD, menopausal status affects the risk of fibrosis severity, with a main effect of early but

not of late menopause. These results, which need validation in independent cohorts, suggest that a more intensive management may be appropriate for females with NAFLD at pre- and early menopause. P818 MICROVESICULAR STEATOSIS; THE MISSED ITEM IN THE MANAGEMENT OF NAFLD? G. Celebi1 , C.N. Ercin1 , A.F. Cicek2 , H. Gurel1 , E. Sertoglu3 , C. Acikel4 , Y. Karslioglu2 , S. Tapan5 , T. Dogru1 , S. Bagci1 . 1 Gastroenterology, 2 Pathology, Gulhane Military Medical Academy, 3 Biochemistry, Mevki Military Hospital, 4 Epidemiology, 5 Biochemistry, Gulhane Military Medical Academy, Ankara, Turkey E-mail: [email protected] Background and Aims: Nonalcoholic fatty liver disease (NAFLD) is among the most common causes of chronic liver disease and ranks as one of the major causes of cirrhosis. In NAFLD, histological course of steatosis is usually macrovesicular, but may be accompanied by varying degrees of microvesicular steatosis. In the present study, we aimed to evaluate the prevalence and significance of microvesicular steatosis in subjects with NAFLD. Methods: A total of 191 nondiabetic male subjects with biopsy proven NAFLD were evaluated retrospectively. Clinical and laboratory data were registered. Hematoxylin-eosin and Masson trichrome stains were evaluated by single expert liver pathologist. Steatosis grade (0–3), steatosis type (macrovesicular, microvesicular or mixed), zone (1, 3, azonal, or panacinar) and other histological findings (Megamitochondria, Mallory–Denk body and Councilman body) were determined. Results: Microvesicular steatosis was found in 30.4% of the participants. There was no difference regarding the demographic and biochemical parameters between patients with or without microvesicular steatosis. On the other hand, the prevalence of ballooning injury and megamitochondria were higher in patients with microvesicular steatosis (p = 0.019 and p = 0.036, respectively). Moreover, in multivariate analysis, the presence of ballooning injury and megamitochondria were significantly associated with microvesicular steatosis (p = 0.005 and p = 0.037, respectively). Conclusions: In conclusion, microvesicular steatosis might be an important histological finding for the prediction of liver injury in NAFLD. Further longitudinal studies are needed to characterize the role of microvesicular steatosis in natural history of NAFLD. P819 PREDICTIVE FACTORS OF SIGNIFICANT STEATOSIS IN POTENTIAL HEALTHY LIVER DONORS D. Jothimani1 , M. Srivatsav1 , S. Reddy1 , M. Rela1,2 . 1 Institute of Liver Disease and Transplantation, Global Hospitals, Chennai, India; 2 King’s College London – Institute of Liver Studies, King’s College Hospital, London, United Kingdom E-mail: [email protected] Background and Aims: Presence of significant (>10% macrovesicular steatosis) fat in potential liver donors may preclude them from partial liver donation. Aim of the study was to identify factors that predict significant steatosis in these patients. Methods: In this retrospective study all potential donors had screening blood tests followed by a computerized tomography (CT) scan. Patients underwent a liver biopsy for one or more of the following: abnormalities in LFTs, lipid profile or liver attenuation index (LAI) 10% steatosis (group 2). Univariate analysis showed significant difference in donor weight (66 vs 72.5 kg, p = 0.04), total cholesterol (193 vs 221 mg/dl, p = 0.02), LDL cholesterol (32.8 vs 38.9 mg/dl, p = 0.079), total liver volume (1301 vs 1393 ml, p = 0.058) and LAI (6.4 vs 1.0, p = 0.002) between group 1 and group 2, respectively. In the multivariate analysis, only total cholesterol (p = 0.042) and LAI (p = 0.02) independently predicted significant steatosis. Conclusions: In this study, total cholesterol and LAI are independent predictive factors of significant steatosis in potential liver donors. This result may be an useful non-invasive tool in patients with non-alcoholic fatty liver disease. P820 A RANDOMIZED CONTROLLED TRIAL TO STUDY THE EFFICACY OF COMBINATION OF PENTOXIPHYLLINE AND VITAMIN E VERSUS VITAMIN E IN PATIENTS WITH NON-ALCOHOLIC STEATOHEPATITIS C.K. Kedarisetty1 , A. Bhardwaj2 , A.S. Bhadoria2 , C. Bihari3 , A. Rastogi3 , U. Kanal4 , S.K. Sarin1 . 1 Hepatology, 2 Research, 3 Pathology, 4 Nutrition, Institute of Liver and Biliary Sciences, New Delhi, India E-mail: [email protected] Background and Aims: Nonalcoholic steatohepatitis (NASH) is the most prevalent cause of chronic liver disease. PIVENS trial showed improved NAS score but not fibrosis with Vitamin E (VE). We have earlier shown (Satapathy et al, JGH 2007) fibrosis regression with pentoxiphylline (PTX). Aim: To study the efficacy of combination of PTX and VE versus VE alone in histology, clinical and biochemical parameters of NASH. Methods: Consecutive histologically proven patients with NASH were randomized to either PTX 400 mg thrice daily and VE 400 IU twice daily (Group A, N = 58) or VE alone (Group B, N = 58). Both were given same diet and lifestyle modifications. Repeat biopsy was done at 1 year in a proportion of patients. Paired biopsies were independently assessed by two blinded pathologists. 3 monthly follow-up done. Results: Both groups were well matched in baseline characteristics. Primary end point was histological improvement at 1 year. I2 patients had repeat biopsy (7 in group A, 5 in group B). No difference in reduction of NAS score noted between both groups, but there was significant fibrosis regression in Group A compared to Group B (p 0.003). Secondary end points namely reduction in body mass index, waist circumference, ALT, APRI, FLP, uric acid, HOMA-IR and fibroscan from baseline to each follow up, were statistically significant in both groups. The reduction in TNF-a levels was statistically significant in Group A compared to Group B. Conclusions: This is the first randomized open label trial (NCT01384578) showing improved efficacy of PTX and VE in regression of fibrosis compared to VE alone with comparable clinical and biochemical improvement. P821 NOVEL MRI TECHNIQUES FOR THE ASSESSMENT OF NON-ALCOHOLIC FATTY LIVER DISEASE M. Pavlides1,2 , R. Banerjee1,3 , E.M. Tunnicliffe1 , J. Collier2 , L.M. Wang4 , K. Fleming4,5 , S. Neubauer1 , E. Barnes2,6 . 1 Oxford Centre for Clinical Magnetic Resonanace Research, 2 Translational Gastroenterology Unit, University of Oxford, 3 Perspectum Diagnostics, 4 Histopathology, John Radcliffe Hospital, 5 Division of Medical Sciences, 6 Peter Medawar Building, University of Oxford, Oxford, United Kingdom E-mail: [email protected] Background and Aims: Magnetic Resonance techniques are increasingly being used in the assessment of liver disease. In this study we aimed to establish whether a novel MRI protocol could S344

be used to diagnose steatohepatitis and stage fibrosis in patients with NAFLD. Methods: 45 patients who had clinically indicated liver biopsies for NAFLD assessment were invited to have a multi-parametric MRI scan. MRI data were collected and analysed prospectively prior to liver biopsy. Liver histology was assessed according to clinical protocols (Ishak stage for fibrosis; NAS score for steatosis and steatohepatitis). The multi-parametric MR protocol included T1 mapping and T2* mapping (for iron quantification) which were used to calculate the iron-corrected T1 (cT1, ms), which measures liver extracellular fluid (increases in fibrosis and inflammation) and proton MR spectroscopy (MRS) for liver lipid quantification. Results: There was a significant correlation between the MRI measures (cT1) of fibrosis and the Ishak stage (rs = 0.57, p < 0.0001). In patients with mild fibrosis (n = 26; Ishak stage F0–2) MRI measures of inflammation (cT1) could distinguish patients with a NAS score ≥5 (median cT1 = 947 ms) from patients with NAS < 5 (median cT1 = 799 ms; p = 0.0006; Figure 1a). The area under the ROC curve for the identification of steatohepatitis was 0.91 (95% CI: 0.78–1.0). There was also a significant correlation between the degree of steatosis measured by proton MR spectroscopy and cT1 (r = 0.63, p = 0.0006; Figure 1b).

Figure 1.

Conclusions: Our multiparametric MRI protocol is a promising method for the assessment of NAFLD as it can assess multiple aspects of the disease quantitatively and non-invasively. P822 MULTIPARAMETRIC MAGNETIC RESONANCE SHOWS THAT OBESITY IS STRONGLY ASSOCIATED WITH HEPATIC STEATOSIS IN ADULTS AND CHILDREN AND CAN DIFFERENTIATE PATIENTS WITH NASH FROM NAFLD R. Banerjee1,2 , B. Rial1 , M. Pavlides1,3 , C. Osuchukwu4 , A.J. Lewandowski1 , J.D. Collier3 , J.E. Schneider1 , S. Neubauer1 , E. Barnes3,5 . 1 Oxford Centre for Clinical Magnetic Resonanace Research, University of Oxford, 2 Perspectum Diagnostics, 3 Translational Gastroenterology Unit, University of Oxford, Oxford, 4 Department of Child Health, Central North West London NHS Trust, Milton Keynes, 5 Peter Medawar Building, University of Oxford, Oxford, United Kingdom E-mail: [email protected] Background and Aims: NAFLD affects 20–35% of adults and children in the UK, and is increasing with the rise in obesity. The aim was to develop a clinical test for NASH in children, using adult reference data. Methods: Multiparametric MR was performed on 201 subjects – 148 adults of varying BMI but no other cardiovascular risk factors, eleven lean boys (BMI within interquartile range, age 10–15), eleven obese boys (BMI >95th centile, age 10–15), and prospectively in 31 patients with subsequent biopsy-proven NAFLD or NASH. Proton spectroscopy measured hepatic lipid content. Iron-corrected T1 relaxation time (cT1) was measured in the biopsy patients and children to estimate hepatic extracellular fluid (ECF), which rises


POSTERS with either inflammation or fibrosis. In the 31 biopsy patients, the histological NAFLD Activity Score (NAS) determined patients with NAFLD (NAS < 5) from those with NASH (NAS ≥ 5). Results: Increasing BMI in adults and children was associated with increasing hepatic steatosis (Figure 1). Obese boys had markedly higher liver fat compared to lean boys (median 2.79 v 0.46; p = 0.04).

support coffee’s protective role in NAFLD is lacking. We tested the association between coffee consumption and NAFLD in a prospective general population cohort. Methods: The analysis was performed both in a cross sectional manner (n = 347, 31% NAFLD) and in a prospective manner in a subcohort without NAFLD at baseline followed for 7 years (n = 147, 19% incident NAFLD). NAFLD was diagnosed with abdominal ultrasound and SteatoTest. FibroTest was used to assess fibrosis. A detailed structured questionnaire on coffee consumption was administrated during a face-to-face interview. Results: NAFLD patients did not differ from controls in their coffee consumption. Subjects with high coffee consumption (of ≥3 cups/day) did not differ from those with lower consumption in steatosis assessed by Hepato-Renal index and SteatoTest, or fibrosis assessed by FibroTest or liver enzymes. In a multivariate analysis, adjusting for age, gender, smoking, sugar consumption and physical activity no association was demonstrated between high coffee consumption and NAFLD (OR = 0.95, 0.60–1.52, 95% CI). In the prospective analysis, no association was demonstrated between high coffee consumption and new onset of NAFLD (OR = 1.02, 0.45–2.35). High coffee consumption was significantly associated with a lower rate of advanced fibrosis (10.4% vs. 26.3%, P = 0.047). Adjusting for the variables mentioned above the association become borderline significant (OR = 0.54, 0.27–1.08, P = 0.083). Conclusions: No association was demonstrated between coffee consumption and NAFLD, but a protective affect from fibrosis is suggested. P824 SOLUBLE CD163, A MACROPHAGE ACTIVATION MARKER, IS INDEPENDENTLY ASSOCIATED WITH STEATOHEPATITIS IN PATIENTS WITH NON-ALCOHOLIC FATTY LIVER DISEASE K. Kazankov1 , F. Barrera2 , H.J. Møller3 , H. Vilstrup1 , J. George2 , H. Grønbæk1 . 1 Department of Hepatology and Gastroenterology, Aarhus University Hospital, Aarhus, Denmark; 2 The Storr Liver Unit, Westmead Millenium Institute, University of Sydney and Westmead Hospital, Westmead, NSW, Australia; 3 Department of Clinical Biochemistry, Aarhus University Hospital, Aarhus, Denmark E-mail: [email protected]

Figure 1. Hepatic lipid content in children, adults, and patients with biopsyproven NAFLD.

Patients with NASH had higher cT1 than those with NAFLD (971±89 ms versus 879±139 ms; p = 0.03). The three obese boys with greatest steatosis also had cT1 ≥970 ms, compared to 783±25 ms in lean boys, suggesting they had steatohepatitis similar to adult NASH. Conclusions: Multiparametric MR can quantify hepatic steatosis, which increases with obesity, in adults and children, and may be used to differentiate NASH from NAFLD. P823 COFFEE DOES NOT PREVENT DEVELOPMENT OF NAFLD BUT MAY DELAY FIBROSIS PROGRESSION: A PROSPECTIVE COHORT STUDY IN THE GENERAL POPULATION S. Zelber-Sagi1,2 , F. Salomone3 , M. Webb1 , H. Yeshua1 , Z. Halpern1,4 , O. Shibolet1,4 . 1 Liver Unit Department of Gastroenterology, Tel Aviv Medical Center, Tel Aviv, 2 School of Public Health, Faculty of Social Welfare and Health Sciences, University of Haifa, Haifa, Israel; 3 U.O.C. di Gastroenterologia, Ospedale di Acireale, Azienda Sanitaria Provinciale di Catania, Catania, Italy; 4 Sackler Faculty of Medicine, Tel Aviv University, Tel Aviv, Israel E-mail: [email protected] Background and Aims: Studies suggest that coffee consumption reduces the risk for type 2 diabetes, cirrhosis, hepatocellular carcinoma and possibly of NAFLD. However, conclusive data to

Background and Aims: Macrophages play an important role in the pathogenesis of non-alcoholic fatty liver disease (NAFLD) and the development of steatohepatitis (NASH). Soluble (s)CD163 is a specific marker of activated macrophages. Methods: In a cross-sectional study we investigated associations between sCD163 and biochemical and histological parameters of liver inflammation and fibrosis in 171 NAFLD patients. Demographic, clinical and biochemical parameters were recorded at the time of the liver biopsy. sCD163 was measured by ELISA. Results: We observed increased sCD163 levels in close association with the severity of liver injury, histologically assessed by the NAFLD Activity Score (NAS) and the fibrosis score. Patients with a histological diagnosis of NASH (NAS ≥ 5) had higher sCD163 compared with those with NAS < 5 [3.8 mg/L (IQR 2.8–5.3) vs. 2.5 mg/L (IQR 1.9–3.4), p < 0.001). sCD163 was associated with biochemical parameters of liver injury and insulin resistance, and showed distinct independent associations with NAS (p = 0.03) and fibrosis stage (p = 0.007) in multivariate regression analyses. Moreover, sCD163 was a predictor of NASH (NAS ≥ 5) independent of other biochemical parameters, and we computed a novel sCD163based predictive score (CD163-NASH) with an area under the Receiver Operating Characteristics curve (AUROC) of 0.83 (95% CI: 0.75–0.91) for the diagnosis of NASH. Conclusions: Levels of sCD163 are increased in association with the severity of liver disease, reflecting macrophage activation in NAFLD and NASH. Soluble sCD163 is associated with NASH independently


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POSTERS of other parameters, and a novel sCD163-based NASH score shows excellent predictive capability of NASH. P825 NON-ALCOHOLIC FATTY LIVER DISEASE IS ASSOCIATED WITH REDUCED SERUM CHOLESTEROL-ADJUSTED VITAMIN E LEVELS D. Pastori1 , M. Del Ben1 , L. Polimeni2 , R. Carnevale1 , C. Nocella1 , C. Calabrese1 , F. Baratta1 , G. Labbadia2 , P. Pignatelli1 , F. Violi1 , F. Angelico2 . 1 Internal Medicine and Medical Specialties, 2 Sapienza University of Rome, Rome, Italy E-mail: [email protected] Background and Aims: Non-alcoholic fatty liver disease (NAFLD) is the commonest liver disease worldwide. No data regarding serum cholesterol-adjusted vitamin E levels in patients with NAFLD are available. Aim of this study was to investigate serum levels of cholesterol-adjusted vitamin E in a large cohort of patients with NAFLD. Methods: We enrolled 310 consecutive patients in which liver steatosis was evaluated with a liver ultrasonographic scanning (US). Severity of NAFLD was defined according to Hamaguchi’s criteria. In 18 patients a biopsy-proven diagnosis of NASH was made. Serum levels of vitamin E were measured and adjusted for serum cholesterol (vit E/chol). Results: Mean age was 53.9 years. NAFLD was found in 81.9% of patients at ultrasonography (US). Subjects with NAFLD (3.4±2.1 mmol/mmol chol) and those with NASH (3.3.±1.8 mmol/mmol chol) showed significantly lower mean values of serum vit E/chol than controls (4.8±2.0 mmol/mmol chol, p < 0.001). After dividing our cohort according to median value of vit E/chol, we found a significant inverse association between US-NAFLD and serum vit E/chol (OR = 0.819 [95% CI 0.727–0.922] p = 0.001 for each point of Hamaguchi score). No significant differences in vitamin E intake was found. In a multivariable logistic regression analysis metabolic syndrome predicted NAFLD (OR = 5,812 [95% CI 2,638–12,805] p < 0.001) whilst vit E/chol was negatively associated (OR = 0.769 [95% CI 0.655–0.904] p = 0.001). Conclusions: Subjects with NAFLD, and not only those with NASH, have significantly reduced serum levels of vit E/chol. Low serum levels of vit E/chol are associated to NAFLD independently from the presence of metabolic syndrome and dietary vitamin E intake. P826 HEPCIDIN LEVELS IN PATIENTS WITH NAFLD WITH OR WITHOUT HYPERFERRITINEMIA AND/OR DYSMETABOLIC IRON OVERLOAD J. Marmur1 , S. Beshara2 , G. Eggertsen2 , N. Albiin3 , O. Danielsson4 , ˚ 1 . 1 Unit of Gastroenterology and Hepatology, R. Hultcrantz1 , P. Stal Dept of Medicine, 2 Unit of Clinical Chemistry, Dept of Laboratory Medicine, 3 Dept of Radiology, 4 Unit of Pathology, Dept of Laboratory Medicine, Karolinska Institute, Karolinska University Hospital Huddinge, Stockholm, Sweden E-mail: [email protected] Background and Aims: Patients with NAFLD often display dysmetabolic iron overload (DIO) or increased serum ferritin without iron overload. In the present study we evaluated serum hepcidin and iron parameters in NAFLD patients having normal or elevated ferritin, with or without DIO, and compared values to those of patients with other chronic liver diseases and healthy controls. Methods: Serum hepcidin was analyzed in 83 patients with chronic liver disease (38 of which had NAFLD) and 39 healthy controls. NAFLD patients were divided into those with normal serum ferritin (NAFLD-N, n = 16), elevated ferritin but normal liver iron (NAFLDFERR, n = 7), or DIO (NAFLD-DIO, n = 15). Liver biopsy was performed in 75/84 patients and hepcidin mRNA in liver was determined with rt-qPCR. S346

Results: Hepcidin levels were increased in HFE-negative hemochromatosis and NAFLD-DIO, and correlated to liver iron stores. In NAFLD, hepcidin correlated to serum ferritin (r2 = 0.20, p < 0.01) and liver iron (r2 = 0.21, p < 0.01) but not to BMI, CRP, NAS-score or steatosis. Patients with NAFLD-DIO had significantly higher transferrin saturation (40±8%) than NAFLD-FERR (25±10%), p < 0.05. There was a good correlation between hepcidin mRNA and serum hepcidin (r2 = 0.46, p < 0.01). Conclusions: In NAFLD, serum hepcidin correlates to iron parameters such as serum ferritin and liver iron in patients either with or without DIO, but not to NAS-score, steatosis or BMI. Transferrin saturation may discriminate between hyperferritinemic NAFLD patients with or without DIO. Apart from genetic hemochromatosis, hepcidin regulation was similar in NAFLD compared to other chronic liver diseases with various degrees of hepatic iron overload. P827 INSULIN SECRETORY FUNCTION AND HBa1c ARE RELATED TO LIVER FIBROSIS IN NONALCOHOLIC FATTY LIVER DISEASE Y. Eguchi1 , Y. Kitajima1,2 , H. Takahashi1 , M. Ono3 , M. Okada1 , H. Hyogo4 , Y. Sumida5 , N. Araki2 , N. Ono2 , T. Eguchi2 , R. Iwakiri1 , K. Anzai1 , JSG-NAFLD. 1 Internal Medicine, Div of Hepatology, Saga, 2 Eguchi Hospital, Ogi, 3 Kochi Medical School, Nangoku, 4 Hiroshima University, Hiroshima, 5 Kyoto Prefectual University of Medicine, Kyoto, Japan E-mail: [email protected] Background and Aims: The relationship between the pathogenesis of diabetes and liver fibrosis (LF) in nonalcoholic fatty liver disease (NAFLD) should be assessed. We conducted a multicenter study to analyze the relationship between LF and HBA1c/insulin secretory function (ISF). Methods: We performed general health check-ups on 10,574 HBsAg and HCV-Ab negative subjects (cohort 1). The relationship between HbA1c and LF evaluated by the Fib4-index was analyzed. Liver biopsies showed 155 NAFLD subjects (cohort 2). HOMA-b was indicatively used for the evaluation of ISF and categorized into 3 groups (Low HOMA-b: C (p.I148M) was genotyped by Taqman assays in 234 patients with CLD (133 men; 180 with non-viral liver diseases; 14–77 years). Liver steatosis was measured non-invasively using controlled attenuation parameter (CAP™) in a subgroup of 97 patients. The control cohort consisted of 279 healthy individuals (114 men; 32–98 years). Results: The PNPLA3 variant increased the odds of presenting with severe liver phenotypes that eventually lead to referral and informed consent for genotyping (common OR = 1.72, P 0.001). Median CAP [dB/m] levels differed significantly (ANOVA P = 0.0025) between carriers of the genotypes [II] (n = 46), [IM] (n = 40) and [MM] (n = 11) and were 246.0, 277.0 and 320.0, respectively. Carriers of the prosteatotic [M] allele had higher (P = 0.0043) median CAP levels as compared to the [II] individuals. Conclusions: This is the first study showing an association between hepatic steatosis quantified using CAP and the PNPLA3 variant across various liver diseases. Our results provide evidence that patients with the PNPLA3 risk variant develop more severe liver diseases, supporting the concept of PNPLA3-associated steatohepatitis (PASH; Semin Liver Dis 2013). P829 LIVER MITOCHONDRIAL FATTY ACID OXIDATION IS ALTERED IN MORBID OBESE PATIENTS WITH NON-ALCOHOLIC STEATOHEPATITIS N. Helmy1,2 , A. Mansouri1,3 , V. Lenoir1 , M. Ziol4 , C. Esnous1 , B. Dousset5 , F. Bouillaud1 , C. Vons1,2 , C. Prip-Buus1 . 1 INSERM U 1016, CNRS UMR 8104, University Paris Descartes, Paris, 2 Service de Chirurgie Digestive et Métabolique, AP-HP, Hˆ opitaux Universitaires Paris-Seine-Saint-Denis, Hˆ opital Jean Verdier, Bondy, France; 3 Institute of Food, Nutrition and Health, ETH Zurich, Zurich, Switzerland; 4 Service d’Anatomo-Pathologie, AP-HP, Hˆ opitaux Universitaires Paris-Seine-Saint-Denis, Hˆ opital Jean Verdier, Bondy, 5 Service de Chirurgie Digestive, Hépato-Biliaire et Endocrinienne, AP-HP, Hˆ opital Cochin, Paris, France E-mail: [email protected] Background and Aims: Non-alcoholic fatty liver disease (NAFLD), ranging from simple steatosis to steatohepatitis (NASH), is often associated with obesity or metabolic syndrome. Whether alterations in liver mitochondrial fatty acid oxidation (FAO) capacity contribute to NASH development remains to be explored in human. Therefore, we presently investigated in fresh liver biopsies from obese and lean patients the interrelation between mitochondrial FAO flux and NAFLD severity. Methods: Per operative liver biopsies were obtained from morbidly obese patients undergoing bariatric surgery and lean control patients (liver resection). NAFLD severity was evaluated by histological analysis, allowing classification of obese patients into 3 groups: G1 (normal liver), G2 (simple steatosis), G3 (NASH). FAO and esterification fluxes were measured in fresh liver biopsies using [1-14 C]oleate. Results: In this study, 45 obese (age=38.2±2.5 years; BMI=44.06±1.06 Kg/m2 ) and 5 control (age=37.6±6.9 years; BMI= 21.9±1.7 Kg/m2 ) patients were included. The distribution of obese patients was: 11 G1 (24%), 23 G2 (51%) and 11 G3 (24%). Hepatic triacylglycerol content was higher in G2 and G3 while diacylglycerol content was significantly increased only in G3. [114 C]Oleate metabolic orientation (oxidation versus esterification) and its oxidation rate were similar in obese and control patients. Taking into account the rate of complete (14 CO2 ) versus incomplete (acido-soluble products, ASP) FAO, we observed a significant decrease in 14 CO2 production and CO2 /ASP ratio in G3 obese patients when compared to other groups (p < 0.05).

Conclusions: These results suggest that NASH patients have a lower liver capacity to completely oxidize fatty acids in comparison to patients with histologically normal or simple steatosic liver. P830 THE −11391G/A POLYMORPHISM IN THE PROMOTER REGION OF THE ADIPOQ GENE PROTECTS FROM SEVERE STEATOSIS IN NONALCOHOLIC FATTY LIVER DISEASE M.J. Brochado1 , F.A. Domenici1 , A.L. Martinelli1 , A.C. Teixeira1 , L.N. Ramalho2 , H.D. Vannucchi1 . 1 Medicine, 2 Pathology, School of Medicine of Ribeirao Preto-USP, Ribeirao Preto, Brazil E-mail: [email protected] Background and Aims: Nonalcoholic fatty liver disease (NAFLD) includes a wide spectrum of liver disease ranging from pure steatosis to nonalcoholic steatohepatitis (NASH) and cirrhosis. Genetic association studies have demonstrated that single nucleotide polymorphisms (SNPs) in AdipoQ gene confer susceptibility for type 2 diabetes and obesity. Moreover, hypoadiponectinemia might represent an independent risk factor for NAFLD. We evaluated serum adiponectin levels in NAFLD patients and investigated the association between the ADIPOQ gene SNPs [−11391G/A, −11377C/G, +45T/G, +276G/T] and susceptibility to NAFLD and severity of hepatic steatosis. Methods: A total of 138 biopsy-proven NAFLD patients (118 NASH and 20 pure steatosis) and 138 healthy controls were recruited. Hepatic steatosis was graded as moderate (≤33%) or severe (>33%). Serum adiponectin was measured by EnzymeLinked Immunosorbent Assay. Four SNPs at the AdipoQ gene were genotyped using allelic discrimination TaqMan method. Results: Serum adiponectin levels were lower in NAFLD than in controls (P < 0.0001, 1.69±1.67 vs. 10.8±5.0 mg/mL). No difference was observed in adiponectin levels comparing NASH and pure steatosis (P = 0.37). No associations were found between ADIPOQ gene SNPs and NAFLD. However, allele A of the −11391G/A polymorphism in NAFLD patients was associated with protection from severe steatosis (P = 0.01, OR = 0.25, 95% CI: 0.088–0.725). Lower adiponectin levels were observed in severe steatosis compared to moderate steatosis (P = 0.008; 1.29±1.0 vs. 2.5±2.3 mg/mL). Conclusions: The −11391G/A polymorphism in promoter region of AdipoQ gene was associated with protection from severe steatosis whilst lower serum adiponectin was associated with severe steatosis. Adiponectin SNPs might play a role in the severity of NAFLD. P831 MAGNETIC RESONANCE IMAGING ALLOWS QUANTITATION OF HEPATIC TRIGLYCERIDE CONCENTRATION IN HUMANS 1 R. Jimenez-Ag ´ uero ¨ , J.I. Emparanza2 , A. Beguiristain1 , L. Bujanda1 , 5 ´ , J.M. Alustiza3 , E. Garc´ıa4 , E. Hijona1 , L. Gallego1 , J. Sanchez M. Perugorria1 , J. Asensio1 , S. Larburu1 , M. Garmendia6 , M. Larzabal6 , M. Portillo7 , L. Aguirre7 , J. Banales1 . 1 Department of Liver and Gastrointestinal Diseases, Biodonostia Research Institute, Donostia University Hospital, University of the Basque Country (UPV), 2 Clinical Epidemiology Unit, CASPe, CIBER-ESP, Biodonostia an, 4 Osatek SA, Research Institute – HUD, 3 Osatek SA, San Sebasti´ San Sebastian, 5 MR Clinical Scientist, Philips Healthcare Iberia, an, Madrid, 6 Department of Pathology, Hospital Donostia, San Sebasti´ 7 Department of Nutrition and Food Science, Faculty of Pharmacy, UPV/EHU, CIBER-obn, Vitoria, Spain E-mail: [email protected] Background and Aims: Non-alcoholic fatty liver disease (NAFLD) is caused by abnormal accumulation of lipids within liver cells. Its prevalence is increasing in developed countries in association with obesity, and it represents a risk factor for non-alcoholic steatohepatitis (NASH), cirrhosis and hepatocellular carcinoma.


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POSTERS Since NAFLD is usually asymptomatic at diagnosis, new noninvasive approaches are needed to determine the hepatic lipid content in terms of diagnosis, treatment and control of disease progression. Here, we investigated the potential of magnetic resonance imaging (MRI) to quantitate the hepatic triglyceride concentration in humans. Methods: Prospective study from 129 adult patients (97 with morbid obesity and 32 controls) that compares multi-echo MRI fat fraction, grade of steatosis estimated on a 0–3 scale by histopathology, and biochemical measurement of hepatic lipid concentration (i.e. Folch value). Results: MRI fat fraction positively correlates with both grade of steatosis (r = 0.77, r2 = 0.60; p < 0.0001) and Folch value (r =0.90, r2 =0.81; p < 0.0001). In this regard, steatosis grades also positively correlate with Folch values (r = 0.71, r2 = 0.50; p < 0.05); however, no differences in the hepatic lipid concentration were found between grades 0 and 1 of steatosis estimated by histopathology. These data resulted in a novel equation to predict the hepatic lipid concentration from MRI fat fractions. Validation of this formula in 31 additional patients resulted in strong correlation between the measured and estimated Folch values (r2 = 0.98; p < 0.0001). Conclusions: Multi-echo MRI is an accurate approach to determine the hepatic lipid concentration, representing a low-cost noninvasive method to diagnose and monitor steatosis in humans. P832 CLINICAL VALIDATION OF THE FLIP ALGORITHM AND SAF SCORE IN NON-ALCOHOLIC FATTY LIVER DISEASE F. Nascimbeni1 , P. Bedossa2 , L. Fedchuk3 , R. Pais3 , P. Lebray3 , T. Poynard3 , V. Ratziu3 . 1 Department of Biomedical, Metabolic and Neural Sciences, University of Modena and Reggio Emilia, Modena, Italy; 2 Pathology Department, Beaujon Hospital, University Paris Diderot, 3 Department of Hepatology and Gastroenterology, Pitié Salpêtrière Hospital, University Pierre et Marie Curie, Paris, France E-mail: [email protected] Background and Aims: Histological classifications of NAFLD are based on morphology, thus empirical, with undetermined clinical correlates and relevance. We investigated whether different histological categories reflect distinct patient profiles and are predicted by relevant clinical/biological features. Methods: 140 patients with suspected NAFLD were reassessed with the new FLIP algorithm and SAF score (S = Steatosis, A = Activity, the sum of ballooning and lobular inflammation, F = Fibrosis; Hepatology, 2012;56:1751) by a single pathologist, blinded to clinical data. Severe disease was defined as A ≥ 3 and/or F ≥ 3. Results: The FLIP algorithm diagnosed 60 (43%) pts with NASH and 63 (45%) with NAFL (non-NASH NAFLD). Patients labeled NASH had more frequently central obesity, diabetes, metabolic syndrome, higher BMI, waist circumference, HOMA-IR, HbA1c than those labeled non-NASH. They also had higher AST, ALT and GGT and lower albumin and platelet count. NASH patients had more often bridging fibrosis (48% vs. 1%, p < 0.001). Central obesity (OR, 5.36; 95% CI, 1.09–26.43), HOMA-IR ≥2 (6.24; 1.66– 23.38) and ALT ≥2ULN (3.46; 1.50–7.94) independently predicted the diagnosis of NASH by the FLIP algorithm. Histologically severe disease was diagnosed in 47 patients (34%), a single one not labeled NASH, but with a SAF score (S0A1F4) suggestive of burnedout steatohepatitis. Independent predictors of histologically severe disease were: female sex (OR, 3.29; 1.02–10.7), HOMA-IR ≥2 (6.23; 1.14–34), ALT ≥2ULN (7.68; 2.60–23), platelet count (0.98; 0.97– 0.99) and serum albumin (0.82; 0.68–0.99). Conclusions: Patients diagnosed with NASH or histologically severe disease by the FLIP algorithm and SAF score have a distinct clinical profile compatible with more advanced insulin resistance and biochemical liver injury. S348

P833 QUERCETIN MODULATES LIPIDS DROPLETS IN Huh7 CELLS BEARING PNPLA3 GENOTYPE GG A. Rojas1 , I. Rancha1 , M. Garc´ıa-Valdecasas1 , J.A. Del Campo1 , S. Clement2 , F. Negro3 , M. Romero-Gomez1 . 1 UCM Digestive Diseases and CIBERehd, Valme University Hospital, Seville, Spain; 2 Division of Clinical Pathology, University Hospital, 3 Division of Clinical Pathology and Gastroenterology and Hepatology, University Hospital, Geneva, Switzerland E-mail: [email protected] Background and Aims: Steatosis refers to an intracellular accumulation of lipids and subsequent formation of lipid droplets (LD) in hepatocytes. PNPLA3 rs738409 genotype (GG) has been strongly associated with the development of steatosis. The aim of our study was to investigate the effect of quercetin on lipid metabolism, LD size and morphology in Huh-7 cells showing pro-steatotic genotype of PNPLA3 (GG). Methods: Huh-7 cells were treated with 50 and 100 mM quercetin for 72h. The morphology of LDs stained with Oil Red O was assessed by immunofluorescence using confocal microscopy. Image analysis was performed using the MetaMorph Software. Lipid metabolismrelated gene expression levels were analysed by RT-PCR and cell protein levels were evaluated by western-blot. Results: LD morphology (area, radium, and volume) was modified by quercetin treatment: [LD, area (mm2 ): Huh7:142.56±34.68; Huh7+Quercetin 50mM:84.87±38.36 (p < 0.03); Huh7+ Quercetin 100mM:77.85±12.17 (p < 0.002); radium (mm): Huh7: 6.71±0.82; Huh7+ Quercetin 50mM: 5.46±0.82 (p < 0.03); Huh7+ Quercetin 100mM: 4.96±0.42 (p < 0.001); volume (mm3 ): Huh7: 1295.1±466.9; Huh7+ Quercetin 50mM: 629.59±374.3 (p < 0.03); Huh7+ Quercetin 100mM:520.87±123.28 (p < 0.003)]; Gene expression levels were modulated by quercetin (DGAT1: Huh7: 1±0.07; Huh7+Q50mM: 0.5±0.2; FASN: Huh7: 1±0.13; Huh7+Q50mM: 0.21±0.09; MTP: Huh7: 1±0.4; Huh7+Q50mM: 0.345±0.23; PPAR-g: Huh7: 1±0.16; Huh7+Q50mM: 1.28±0.02; APOB: Huh7: 1±0.15; Huh7+Q50mM: 0.18±0.3; PNPLA3: Huh7: 1±0.05; Huh7+Q50mM: 0.4±0.06; ACC: Huh7: 1±0.07; Huh7+Q50mM: 0.39±0.4; SREBP: Huh7: 1±0.08; Huh7+Q50mM: 0.52±0.13).

Figure: Huh7 cells treated with quercetin 50 and 100mM.

Conclusions: Quercetin modifies LD morphology. Lipid metabolismrelated gene and protein were modified by quercetin in a dose dependent manner. This flavonoid may interfere in the development of NAFLD in presence of unfavourable genotype GG of PNPLA3.


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Figure (abstract P834).

P834 NON-INVASIVE MANAGEMENT OF NON-ALCOHOLIC FATTY LIVER DISEASE R. Gallego-Duran ´ 1 , P. Cerro-Salido2 , M.J. Pareja3 , I. Ranchal1 , M.C. Rico1 , L. Rojas1 , R. Aznar4 , E. Bugianesi5 , J. Crespo6 , M.T. Arias-Loste6 , J. Abad7 , S. Soto8 , R. Aparcero1 , I. Moreno9 , ´ alez ´ 2, J.L. Calleja7 , O. Lo Iacono8 , R. Andrade9 , E. Gomez-Gonz 1 1 . UCM Digestive Diseases & CIBERehd, Valme M. Romero-Gomez ´ University Hospital, 2 Physics Interdisciplinary Group, University of Sevilla, 3 Pathology Unit, 4 Radiology Unit, Valme University Hospital, Sevilla, Spain; 5 Turin University, Turin, Italy; 6 Marques de Valdecilla University Hospital, Santander, 7 Puerta de Hierro Hospital, 8 Tajo Hospital, Madrid, 9 UCM of Digestive Diseases & CIBERehd, Virgen de la Victoria University Hospital, M´ alaga, Spain E-mail: [email protected] Background and Aims: To evaluate non-invasive management of NAFLD using biochemical markers and an image-based method DEMILI® tool for the staging of fibrosis and diagnosis of steatohepatitis. Methods: Sixty-eight patients who underwent percutaneous liver biopsy by NAFLD clinical suspicious were included. All these

patients underwent magnetic resonance study, mean age 51±12 y, 45 (66.6%) males. SAF Score, NAS Score, and fibrosis stage were reported by a single pathologist. Significant fibrosis (F2F3F4) was found in 25/68 (37%) of patients, and 33/68 (49%) showed steatohepatitis. FGF-21 and CK-18 were measured in 12 hoursfasting sera of patients using ELISA kits, and APRI, FORNS, FIB-4 and PNPLA3 rs738409 genotype were also determined. Diagnostic accuracy was assessed by AUROC using SPSS v22.0. Results: DEMILI® software showed an AUROC of 0.85 (95% CI: 0.74–0.97) for significant fibrosis and 0.89 (95% CI: 0.78–1.0) for steatohepatitis. FGF-21 levels showed an AUROC of 0.794 (95% CI: 0.633–0.954) for significant fibrosis detection and 0.688 (95% CI: 0.498–0.879 for steatohepatitis prediction. Remaining methods yielded an AUROC lower than 0.70. CC-PNPLA3 genotype patients showed less rates of steatohepatitis and lower score in NASHMRI (0.47±0.27 CC-genotype vs 0.67±0.29 CG/GG-genotype; p = 0.04). No differences were seen in FGF21, CK18, Forns, APRI or FIB-4. Conclusions: DEMILI® , but not biochemical markers, seems to accurately predict steatohepatitis. Fibrosis stage was predicted by DEMILI® and FGF21. Further analyses and algorithms for stage fibrosis prediction including both methods are warranted.


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POSTERS Acknowledgements: The research leading to these results has received funding from the European Community’s Seventh Framework Programme (FP7/2007–2013) under grant agreement no. HEALTH-F2–2009–241762 for the project FLIP. P835 LIVER STEATOSIS ASSESSMENT WITH CAP AT 3.5 MHz USING FIBROSCAN M AND XL PROBE V. De Ledinghen1 , C. Corpechot2 , J. Vergniol1 , P. Bedossa3 , O. Chazouilleres2 , Y. Menu4 , V. Paradis3 . 1 Hepato-Gastroenterology, Haut Leveque Hospital, Pessac, 2 Hepato-Gastroenterology, Saint Antoine Hospital, Paris, 3 Pathology, Beaujon Hospital, Clichy, 4 Radiology, Saint Antoine Hospital, Paris, France E-mail: [email protected] Background and Aims: Controlled Attenuation Parameter (CAP) measures liver ultrasound attenuation at 3.5 MHz using FibroScan (FS). Currently available on the M probe to quantify steatosis, the aim of this work was to implement the measurement of CAP on the XL probe (dedicated to overweighed patients) and validate its diagnosis performance. Methods: 180 consecutive patients (21% NAFLD, 25% VHC, 14% VHB, and 40% other – BMI=25±11 kg/m2 , age=51±15years, 53% male) with histological assessment of steatosis within 7 days of FS (liver biopsy≥15 mm, double-blind reading) were enrolled. Steatosis was graded and distributed as follows: S0: steatosis≤10% (63%), S1: 11–33% (11%), S2: 34–66% (10%), S3: ≥67% (16%). Spearman correlation coefficient (SCC), intra-class correlation coefficient (ICC), Area under Receiver Operating Characteristic curve (AUROC) and Delong-test for comparison of AUROC were used to analyse the data. Results: SCC of CAP with histological steatosis was 0.65 and 0.64 for XL and M probes respectively. The AUROCs (area under ROC curve) of CAP at 3.5 MHz versus histological steatosis (Table 1) were not significantly different when measured by either the M or the XL probe and cut-off maximizing the Youden index for the M probe had similar performance for CAP measured with XL probe.

P836 ELEVATED SERUM LEVELS OF ALT ARE ASSOCIATED WITH METABOLIC SYNDROME INDEPENDENT OF FATTY LIVER DIAGNOSED BY ULTRASOUND Y. Imamura1 , H. Uto2 , Y. Hiramine1 , K. Hosoyamada1 , S. Yoshifuku3 , H. Miyahara3 , S. Maenohara3,4 , M. Oketani2 , A. Ido2 , H. Tsubouchi5 . 1 Department of Internal Medicine, Kagoshima Kouseiren Hospital, Kagoshima, 2 Digestive and Lifestyle Diseases, Department of Human and Environmental Sciences, Kagoshima University Graduate School of Medical and Dental Sciences, 3 Kagoshima Kouseiren Medical Health Care Center, 4 Department of Surgery, Kagoshima Kouseiren Hospital, 5 Department of HGF Tissue Repair and Regenerative Medicine, Kagoshima University Graduate School of Medical and Dental Sciences, Kagoshima, Japan E-mail: [email protected] Background and Aims: Elevated levels of alanine aminotransferase (ALT), a surrogate marker of liver injury, are mainly due to fatty liver (FL) and are considered an indicator of metabolic syndrome (MS). However, limited information is available on the influence of FL on this association between elevated ALT and MS. Methods: Individuals receiving medical examinations who are negative for both HBs-antigen and HCV antibody were enrolled (6459 men and 4487 women). ALT >30 IU/L was considered elevated. FL was diagnosed using ultrasonography (US-FL). Results: There were 1778 (27.5%) men and 387 (8.6%) women with elevated ALT. Logistic regression analysis revealed that obesity (odds ratios [95% confidence interval] 2.09 [1.77–2.45] among men and 2.31 [1.72–3.13] among women), dyslipidaemia (1.90 [1.67–2.17] and 1.88 [1.42–2.40]), hypertension (1.33 [1.16–1.52] among men), and US-FL (3.34 [2.92–3.84] and 3.91 [3.00–5.12]) were independent predictors of elevated ALT. It is noteworthy that among the subjects with elevated ALT, approximately one-third (578 men and 148 women) did not have US-FL. The risk of elevated ALT increased with the number of MS components in both of US-FL(−) and US-FL(+) subjects (Figure 1).

Table 1.

Steatosis ≥11% (n = 66, 37%) AUROC ± SD Cut-off Sensitivity Specificity Steatosis ≥34% (n = 47, 26%) AUC ± SD Cut-off Sensitivity Specificity Steatosis ≥67% (n = 29, 16%) AUC ± SD Cut-off Sensitivity Specificity

M Probe

XL Probe

0.832±0.034 267 dB/m 68.2 86.0

0.836±0.033

0.895±0.032 272 dB/m 82.9 87.9 0.945±0.017 290 dB/m 86.2 84.7

66.7 89.5 0.900±0.031 82.9 87.2 0.949±0.019 89.7 86.8

Conclusions: Measurement of CAP at 3.5 MHz is feasible in with the FS XL probe and therefore in overweighed patients for whom steatosis is more prevalent. This work confirms that the diagnosis accuracy of CAP for steatosis assessment is equivalent when using either the M or the XL probe.

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Figure 1. Risk of ALT abnormality.

Conclusions: There is an independent association between the risk of elevated ALT and the number of MS components regardless of the presence of US-FL, suggesting the importance of MS in the pathogenesis of liver injury. P837 GENOTYPE 3 AND CIRCULATING SCCA-IgM ARE INDEPENDENTLY ASSOCIATED WITH HISTOLOGICAL FEATURES OF NASH IN HCV INFECTED PATIENTS A. Martini1 , G. Fattovich2 , M. Guido1 , E. Bugianesi3 , A. Biasiolo1 , D. Ieluzzi4 , A. Gallotta5 , G. Fassina5 , C. Merkel1 , A. Gatta1 , F. Negro6 , P. Pontisso1 . 1 Dept of Medicine, University of Padua, Padua, 2 Dept of Medicine, University of Verona, Verona, 3 Division of Gastroenterology and Hepatology, Dept of Medical Sciences, University of Turin, Turin, 4 Division of Gastroenterology, Azienda Ospedaliera Universitaria Integrata, Verona, 5 Xeptagen, VEGA Park, Venice, Italy; 6 Divisions of Gastroenterology and Hepatology and of Clinical Pathology, University Hospitals, Geneva, Switzerland E-mail: [email protected] Background and Aims: In chronic HCV infection liver steatosis is a frequent feature, especially in genotype 3, but its clinical


POSTERS significance is still debated. Since SCCA-IgM immunocomplexes have been associated with more advanced liver disease and increased risk of HCC development, the purpose of this study was to evaluate the occurrence of this biomarker and its possible relation with the presence of NASH at liver biopsy. Methods: In 91 patients with biopsy proven chronic hepatitis C serum samples were tested for SCCA-IgM by ELISA. Sera of 92 consecutive HCV negative patients with histological NAFLD were included as controls. Results: SCCA-IgM was detected in 33% of HCV patients at the time of liver biopsy, but only in 4% of patients with NAFLD. In chronic hepatitis C this biomarker was found more elevated in patients with concomitant histological features of NASH and at multivariate analysis SCCA-IgM and genotype 3 were independently associated with this histologic feature. Referred to NASH, specificity and sensitivity were 97% and 44% for HCV genotype 3 vs 95% and 26% for SCCA-IgM, while PPV and PNV were 80% and 86% for the former vs 70% and 73% for the latter variable. After antiviral treatment, in HCV patients with SVR, SCCA-IgM values decreased significantly during treatment and remained persistently low after the end of therapy, while remained unchanged in the other patients. Conclusions: SCCA-IgM was detectable in about one third of patients with chronic hepatitis C and its presence was significantly associated with histological NASH, independently of the infecting genotype. P838 DISTURBED ENERGY METABOLISM AND LIPID PROFILE AFTER A 4 WEEK HIGH ORAL FRUCTOSE CHALLENGE: POSSIBLE ROLE OF BILE ACIDS C. Kienbacher1,2 , S. Traussnigg1 , E. Halilbasic1 , C.D. Fuchs2 , W. Dolak1 , P. Steindl-Munda1 , T. Stojakovic3 , G. Fauler3 , T. Claudel2 , M. Trauner1,2 . 1 Division of Gastroenterology and Hepatology, Department of Internal Medicine III, 2 Hans Popper Laboratory of Molecular Hepatology, Division of Gastroenterology and Hepatology, Department of Internal Medicine III, Medical University of Vienna, Vienna, 3 Clinical Institute of Medical and Chemical Laboratory Diagnostics, Medical University of Graz, Graz, Austria E-mail: [email protected] Background and Aims: Fructose overconsumption in Western-diet is linked to non-alcoholic fatty liver disease (NAFLD) and impaired glucose and lipid metabolism. Since bile acids (BA) are critical regulators of lipid and energy metabolism, we aimed to explore whether dietary fructose challenge (FC; 150 g/day for 4 weeks) may impact on lipid and glucose dysbalance by changing BA metabolism. Methods: Ten healthy volunteers were enrolled (m:f = 5:5; age = 24.5 [21–37] years; p = n.s.). Blood was collected prior and after FC for routine laboratory analysis, BA and lipid profiling. Hepatic fat content was measured using magnetic-resonance-spectroscopy (MRS). Results: FC increased BMI (mean±SD; 21.11±2.68 vs 21.51±2.77 kg/m2 ; p < 0.001) and fasted serum glucose (83.2±8.37 vs 88.4±5.48 mg/dL; p = 0.035), indicating changes in energy metabolism. Although no changes were observed in total serum cholesterol and triglyceride levels; VLDL-cholesterol (17.2±6.05 vs 11.5±7.29 mg/dL; p = 0.034), -cholesterol-ester (8 [2–14] vs 3 [1–11]mg/dL; p = 0.025), and -ApoB (8.5 [2–16] vs 6 [1–12] mg/dL; p = 0.039) were reduced after FC, reflecting lower hepatic VLDL excretion and/or faster clearance. FC did not change total serum BA concentration that could explain lower VLDL excretion via FXR-activation. However, increased ApoAII levels (43.8±7.48; 49.0±10.76 mg/dL; p = 0.042) may favor faster VLDL clearance. Of interest, FC did not increase total intrahepatic lipids in MRS and serum liver enzymes. The secondary BAs (converted by intestinal bacteria) lithocholic-acid and ursodeoxycholic-acid

tended to decrease under FC (two-sided-p = 0.097 and p = 0.075, respectively). Conclusions: FC induced changes in energy metabolism as reflected by weight gain, increased fasted glucose and changes in VLDL metabolism that may be linked to increased ApoAII levels. Moreover, shifts in BA-composition could be due to alterations in fecal excretion and/or enterohepatic circulation, perhaps by shifts in the intestinal microbiome thus influencing BA-mediated FXR and TGR-5 signalling. Acknowledgements: The research leading to these results has received funding from the Medical Scientific Fund of the Mayor of the City of Vienna (grant number 12040) to MT. P839 HANDLING AN ORAL GLUCOSE LOAD AFTER A 4 WEEK ORAL HIGH FRUCTOSE CHALLENGE AND ITS IMPACT ON FETUIN-A C. Kienbacher1 , D. Lorant2 , S. Traussnigg1 , H. Haslacher3 , P. Steindl-Munda1 , M. Krebs4 , G.H. Schernthaner5 , M. Trauner1 . 1 Division of Gastroenterology and Hepatology, Department of Internal Medicine III, 2 Department of Anaesthesia, Intensiv Care Medicine and Pain Management, 3 Department of Laboratory Medicine and Biobank of the Medical University of Vienna, 4 Division of Endocrinology and Metabolism, Department of Internal Medicine III, 5 Department of Internal Medicine II, Medical University of Vienna, Vienna, Austria E-mail: [email protected] Background and Aims: Fetuin-A is a liver-derived hepatokine which induces insulin resistance (IR) and is associated with non-alcoholic fatty liver disease (NAFLD) and cardiovascular disease (CVD). High fructose consumption is also linked to CVD and development and progression of NAFLD. To test our hypothesis that hepatic Fetuin-A production may be influenced by fructose intake and therefore could represent an important factor in the pathogenesis of NAFLD and CVD, we examined the impact of a high oral fructose challenge (FC; 150 g/day for 4 weeks) on serum Fetuin-A levels. Methods: Ten healthy volunteers were enrolled (m:f = 5:5; age = 24.5 [21–37]; p = n.s.). Fasted serum Fetuin-A, Glucose, Insulin and C-Peptide were assessed prior and after FC and in a frequently sampled (30/60/90/120 min) single oral glucose load (oGTT; 75 g). Results: After FC changes in energy metabolism reflected by increased BMI (21.11±2.68 vs 21.51±2.77 kg/m2 ; p < 0.001) as well as fasting glucose levels (83.2±8.37 vs 88.4±5.48 mg/dL; p = 0.035) were observed. Fasted serum Fetuin-A levels remained unchanged, but increased in the oGTT after FC at 90min (260.9±30.1 vs 277.2±43.3 mg/mL; p = 0.046) and 120 min (234.8 [224.1–332.2] vs 288.1 [238.4–372.5] mg/mL; p = 0.007). Concomitantly, as an early response at 30 min, Insulin (37.95 [2–128.9] vs 58.95 [20.4–163.7] mU/mL; p = 0.059) and C-Peptide (5.7 [4.5–11.6] vs 8 [4.9–12] ng/mL]; p = 0.139) tended to increase. Of note, Insulin (32.4 [7.15–91.79] vs 14.54 [3.61–27.65] mU/mL; p = 0.005), C-Peptide (7.22±3.84 vs 4.41±1.96 ng/mL; p = 0.012) and glucose (105.2±33.38 vs 79.1±14.5 mg/dL; p = 0.035) decreased at 120 min, suggesting healthy young volunteers are still capable of disposing elevated glucose levels after FC, also reflected by unchanged HOMAIR, OGIS-Index and Adipo-IR. Conclusions: Despite unchanged Fetuin-A-levels in fasted conditions, a significant increase in Fetuin-A in response to a single oral glucose load was observed after a long-term high oral fructose challenge. This postprandial perturbance in Fetuin-A-levels could mechanistically link fructose overconsumption to NAFLD and CVD risk with possible therapeutic consequences. Acknowledgements: The research leading to these results has received funding from the Medical Scientific Fund of the Mayor of the City of Vienna (grant number 12040) to MT.


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POSTERS P840 ALTERATION IN LIPID METABOLISM AFTER AN ORAL FAT LOAD IN PATIENTS WITH NAFLD L. Mezzabotta1 , E. Vanni1 , C. Rosso1 , M. Gaggini2 , R. Gambino1 , E. Buzzigoli2 , C. Saponaro2 , D. Ciociaro2 , M.L. Abate1 , F. Saba1 , F. Salomone3 , G.P. Caviglia1 , S. Carenzi1 , A. Smedile1 , M. Rizzetto1 , M. Cassader1 , A. Gastaldelli2 , E. Bugianesi1 . 1 Medical Sciences, University of Turin, Turin, 2 Cardiometabolic Risk Unit, Institute of Clinical Physiology – CNR, Pisa, 3 U.O.C. of Gastroenterology, Azienda Sanitaria Provinciale di Catania, Catania, Italy E-mail: [email protected] Background and Aims: Non-alcoholic fatty liver disease (NAFLD) is associated with insulin resistance (IR) and impairment in whole body lipid metabolism leading to hepatic fat accumulation and fibrosis. We investigated the effect of a lipid load on lipolysis and hepatic glucose metabolism in non-diabetic/non-dyslipidaemic NAFLD patients. Methods: [2 H5 ]glycerol and[2 H2 ]glucose kinetics, plasma levels of glucose/insulin, lipid profile, endogenous glucose production (EGP) and lipolysis (Glycerol_Ra) were determined in 20 patients with biopsy proven NAFLD and 9 controls during an oral fat load (200 ml dairy cream and an egg yolk). Peripheral/hepatic/adipose tissue-IR indices were derived from plasma glucose/insulin, EGP and lipolysis. Subcutaneous, visceral and hepatic fat were assessed by NMR. Results: During fasting, EGP and lipolysis were similar in NAFLD patients and in CT (8.9±1.2 vs 8.7±0.9umol/kg min and 2.4±0.9 vs 2.1±0.6umol/kg min) although indices of IR were higher in NAFLD patients (HOMA2.3±0.9 vs 1.3±0.4, Hep-IR 92±34 vs 52±18, AdipoIR 21±10 vs 11±5 all p < 0.03). After lipid load, the triglycerides (TG) increase in NAFLD was 2-folds than CT (iAUC-TG p < 0.03) while FFA concentration increased similarly. Lipolysis was suppressed in CT but was unchanged in NAFLD, suggesting a remarkable Adipo-IR at low insulin levels, that was directly related to NAS score and degree of fibrosis (p < 0.01 for both). Adipo-IR was increased proportionally to fat accumulation in subcutaneous, visceral and hepatic fat (r2 = 0.163, p < 0.01; r2 = 0.321, p < 0.001; r2 = 0.24, p < 0.001,respectively). Conclusions: The metabolic handling of an oral fat load is impaired in subjects with NAFLD independent of pre-existing diabetes or dyslipidemia and can contribute to the progression of liver damage. Funded by FP7/2007–2013 under grant agreement no. HEALTH-F2– 2009–241762 for the project FLIP and byPRIN-2009ARYX4T. P841 AWARENESS AND OPINION OF NONALCOHOLIC FATTY LIVER DISEASE (NAFLD) PATIENTS ABOUT OBESITY AND ITS CONSEQUENCES S.P. Singh1 , B. Misra1 , D. Misra1 , G.K. Pati1 , A. Singh1 , S.K. Kar1 , M.K. Panigrahi1 , C. Meher2 , O. Agrawal2 . 1 Department of Gastroenterology, S.C.B. Medical College, 2 Department of Radiology, Beam Diagnostics, Cuttack, India E-mail: [email protected] Background and Aims: The prevalence of nonalcoholic fatty liver disease [NAFLD] is rising globally. The aim of this study is to know the opinion of NAFLD patients regarding NAFLD and obesity and its consequences. Methods: This prospective study was conducted in 326 incidentally detected NAFLD patients using a face-to-face questionnaire. Results: Of 326 patients, 68 (20.86%) had normal weight (BMI < 23); 258 (79.14%) were either overweight or obese (BMI > 23). 23.93% didn’t have knowledge about harmful effects of obesity. About half didn’t know that consumption of fast food and prepacked meals promoted obesity. Two thirds (64.42%) had not been advised by their doctors to reduce weight. Besides, 38.76% (100/258) of the obese presumed that they had normal body S352

weight in comparison to lean NAFLD (46/68) [38.76% versus 67.65%; p < 0.0002]. 61.24% of patients who considered themselves obese attributed it to inactivity, overeating, familial and combined causes in 55.06%, 28.48%, 3.8% and 12.03% cases respectively. One third of obese NAFLD [50/158 (31.65%)] didn’t have guilt of being overweight and 9.49% (15/158) didn’t want to lose weight. Conclusions: Despite high prevalence of NAFLD, awareness among NAFLD patients regarding obesity is grossly inadequate. A third of NAFLD patients are ignorant about harmful effects of obesity. Two fifths of obese NAFLD felt that they were non-obese. Two thirds of NAFLD patients were neither educated by their physicians about effects of obesity nor advised to shed weight. There is a gross lack of awareness regarding NAFLD and obesity in patients and physicians. P842 A STUDY OF RISK FACTORS ASSOCIATED WITH NONALCOHOLIC FATTY LIVER DISEASE [NAFLD] IN INDIANS S.P. Singh1 , D. Misra1 , A. Singh1 , B. Misra1 , G.K. Pati1 , M.K. Panigrahi1 , S.K. Kar1 , C. Meher2 , O. Agrawal2 . 1 Department of Gastroenterology, S.C.B. Medical College, 2 Department of Radiology, Beam Diagnostics, Cuttack, India E-mail: [email protected] Background and Aims: To study the risk factors associated with NAFLD in Indians. Methods: 464 consecutive NAFLD patients and 181 controls were subjected to detailed questionnaire regarding risk factors, anthropometric measurements and biochemical assays including blood sugars, LFT, lipid profile and HOMA. Comparison of different variables was made between the NAFLD patients and controls by using SPSS 17. Results: NAFLD patients had higher BMI [26.25±3.80 vs 21.46±3.08 kg/m2 ], waist-hip ratio [0.96± .12 vs 0.90± .08] and waist-height ratio [0.57± .09 vs 0.50± .06] compared to controls. Fasting blood sugar [101.88±31.57 vs 90.87±10.74 mg/dl] and triglyceride level [196.16±102.66 vs 133.20±58.37 mg/dl] were significantly higher in NAFLD group. HOMA-IR was also higher in NAFLD group [2.53±2.57 vs 1.16± .58]. Majority (90.2%) of NAFLD patients were sedentary. Family history of metabolic syndrome was present in 31.2% NAFLD patients compared to 7% controls [p < 0.0001]. Dietary risk factors associated with NAFLD were nonvegetarian diet [35% vs 23%; p = 0.002], fried food [35% vs 9%; p < 0.0001], spicy foods [51% vs 15%; p < 0.001] and tea [55% vs 39%; p < 0.001]. Diabetes and hypertension were commoner in NAFLD. Snoring (57% vs 47%) and sleep apnea syndrome [17% vs 4.4%] were commoner in NAFLD. Conclusions: The risk factors associated with NAFLD in Indians are sedentary lifestyle, family history of metabolic syndrome, consumption of meat/fish, spicy foods, fried foods and tea. Other risk factors associated with NAFLD included snoring and metabolic syndrome. P843 ALTERATION IN GLUCOSE METABOLISM AFTER AN ORAL GLUCOSE LOAD AND RELATIONSHIP WITH LIVER DAMAGE IN PATIENTS WITH NAFLD E. Vanni1 , L. Mezzabotta1 , C. Rosso1 , M. Gaggini2 , R. Gambino1 , F. Salomone3 , E. Buzzigoli2 , C. Saponaro2 , F. Saba1 , D. Ciociaro2 , G.P. Caviglia1 , M.L. Abate1 , S. Carenzi1 , A. Smedile1 , M. Rizzetto1 , M. Cassader1 , A. Gastaldelli2 , E. Bugianesi1 . 1 Medical Sciences, University of Turin, Turin, 2 Cardiometabolic Risk Unit, Institute of Clinical Physiology – CNR, Pisa, 3 U.O.C. of Gastroenterology, Azienda Sanitaria Provinciale di Catania, Catania, Italy E-mail: [email protected] Background and Aims: Non-alcoholic fatty liver disease (NAFLD) is associated with insulin resistance (IR) for different substrates and


POSTERS in different sites. The purpose of this study was to focus on IR for glucose metabolism and its relationship with liver damage. Methods: We used [2 H2 ]glucose to calculate endogenous glucose production (EGP) in the fasting state and during an oral glucose load (OGTT) enriched with U-13C-glucose to further assess glucose absorption and clearance. Twenty-five non-diabetic/nondyslipidaemic patients with biopsy proven NAFLD and 10 controls (CT) were studied. Results: During OGTT, the glucose curves of NAFLD patients were comparable to controls; however, the insulin curves were 1.5-fold higher (AUC-insulin p < 0.05) with a stepwise increase according to the degree of NAS score (p < 0.05) and liver fibrosis (p < 0.01). Intestinal glucose absorption was similar in the two groups. Glucose clearance (GC) was significantly impaired in NAFLD subjects (AUCglucose clearance decreased 2-folds, p < 0.001). However, when we split GC into muscle and adipose tissue components, only muscle GC was defective and directly related to NAS score and fibrosis (p < 0.05 for both) independent of insulin levels. On the other hand, insulin efficiently suppressed EGP in the liver in both groups. Conclusions: In non-diabetic/non-dyslipidaemic NAFLD patients post-load glucose clearance is significantly impaired in the muscle and is associated with liver fibrosis, while insulin action in the liver is still preserved. Muscle IR is a primary defect in NAFLD and contributes to liver damage. Funded by FP7/2007–2013 under grant agreement no. HEALTH-F2– 2009–241762 for the project FLIP and by PRIN 2009ARYX4T. P844 SHORT-TERM EFFECT OF LOW-CALORIE DIET (LCD) VERSUS ISOCALORIE DIET (ICD) ON BLOOD AMINOTRANSFERASES LEVEL AND LIPIDS PROFILE IN PATIENTS WITH NON-ALCOHOLIC STEATOHEPATITIS (NASH) K. Selezneva1 , O. Kirillova2 , I. Vorozhko2 , V.A. Isakov1 , T. Sentsova2 . Gastroenterology & Hepatology, 2 Lab. Biochemistry, Immunology and Allergology, Institute of Nutrition, Moscow, Russian Federation E-mail: [email protected]

1

Background and Aims: Diet modification is widely used for the treatment of NASH. Caloric restriction was shown to be effective in normalizing blood lipid profile, increasing insulin sensitivity and subsequent normalization of aminotrasferases blood level. However, it was not clearly established, what degree of caloric restriction is sufficient to achieve the normalization of aminotrasferases. Methods: 174 NASH patients (f:m = 86:88) were randomly assigned (as 1:2) to LCD (1700–1900 kcal/day) or ICD (2200– 2500 kcal/day) calculated according to patients’ sex, age, resting energy expenditures and daily physical activity. Caloric restriction was achieved by decreasing consumption of carbohydrates and fat in LCD, whereas for ICD the caloric consumption was established according to the recommended daily values for proteins, fat and carbohydrates for ideal BMI for every patient. Blood chemistry was assessed at baseline and after 1 mo of prescribed diets. Compliance for the diet was also evaluated using previously validated questionnaire. LCD group n = 58

ALT, IU/l AST, IU/l CHOL, mmol/l HDL, mmol/l LDL, mmol/l TG, mmol/l

ICD group n = 116

Baseline

1 mo

P

Baseline

1 mo

P

77.2±31.8 52.7±23.6 5.39±1.02 1.24±0.35 3.44±0.84 1.73±0.67

81.2±50.6 52.8±34.7 4.2±1.1 0.94±0.15 2.76±0.91 1.33±0.4

0.75 0.98 0.001 0.001 0.001 0.004

98.8±45.7 49.6±41.5 5.64±1.02 1.21±0.27 3.40±1.01 2.27±1.29

77.5±41.7 38.6±27.3 5.23±0.88 1.14±0.26 3.31±0.86 1.69±0.81

0.001 0.001 0.001 0.02 0.31 0.001

Results: After 1 mo of dietetic interventions ALT and AST levels were decreased in ICD group, but not in LCD group, whereas blood cholesterol, triglycerides, high-density lipoprotein were

significantly decreased in both groups (table). Compliance index was much higher in ICD group than in LCD group (85% vs 54%). Conclusions: LCD was associated with poor compliance and little effect on to blood aminotransferases during short-term dietetic intervention in compare to ICD in NASH patients. However, both types of diets produced similar effect on to blood lipids profile. Diet modification should be reconsidered in NASH patients in favor of fewer calories restricted and easy to follow diet. P845 IMPACT OF FAT VERSUS GLUCOSE LOAD ON OXIDATIVE STRESS IN NON-ALCOHOLIC FATTY LIVER DISEASE C. Rosso1 , E. Vanni1 , L. Mezzabotta1 , R. Gambino1 , F. Saba1 , S. Carenzi1 , A. Gastaldelli2 , M. Gaggini2 , E. Buzzigoli2 , C. Saponaro2 , F. Carli2 , G.P. Caviglia1 , M.L. Abate1 , F. Salomone3 , A. Smedile1 , M. Rizzetto1 , M. Cassader1 , E. Bugianesi1 . 1 Medical Sciences, University of Turin, Turin, 2 Cardiometabolic Risk Unit, Institute of Clinical Physiology – CNR, Pisa, 3 U.O.C. of Gastroenterology, Azienda Sanitaria Provinciale di Catania, Catania, Italy E-mail: [email protected] Background and Aims: In Nonalcoholic Fatty Liver Disease (NAFLD) oxidative stress (ox-stress) and high circulating levels of oxidizedlow density lipoproteins (ox-LDLs) have been involved in the progression to steatohepatitis (NASH). We investigated the pattern and change of ox-stress parameters in NAFLD patients after a glucose vs a lipid load. Methods: Lipid profile, ox-LDLs and total antioxidative status (TAS) were measured during fasting in 54 subjects (45 patients with biopsy proven NAFLD and 9 healthy controls) and after a 240 oral glucose or fat load (glucose75g, 200 ml dairy cream+an egg yolk) in two subgroups of 25 and 20 patients, respectively. Results: During fasting ox-LDLs were significantly higher in NAFLD patients compared to controls (49±12 vs 38±15; p = 0.03), while TAS was similar. In NAFLD patients, ox-LDLs were positively correlated with triglycerides (r2 = 0.0104, p = 0.034), FFA (r2 = 0.095, p = 0.047), and cholesterol profile (all p < 0.01). Among histological features, only steatosis was significantly associated with basal oxLDLs (r2 = 0.094, p = 0.048). After the lipid load, ox-LDLs increased significantly only in patients with fibrosis (F >0) (36±80 vs 53±17, p = 0.024 at 240’) while TAS decreased in patients with steatosis≥33% (296±26vs 38±37, p = 0.01). Notably, ox-LDLs showed a positive correlation with fatty liver index (FLI) both during fasting (r2 = 0.155, p = 0.009) and after lipid load (r2 = 0.471, p = 0.002 at 240’). A similar pattern was also observed after the glucose load. Conclusions: Hepatic ox-stress is part of a more generalized condition of chronic, whole body oxidative stress that can be enhanced by metabolic changes occurring in the postprandial phase. Funded by FP7/2007–2013 under grant agreement no. HEALTH-F2– 2009–241762 for the project FLIP and by PRIN 2009ARYX4T. P846 SERUM FREE FATTY ACID COMPOSITION IS ASSOCIATED WITH DIFFERENT DEGREES OF FIBROSIS R. Gambino, N. Alemanno, S. Pinach, F. Saba, L. Mezzabotta, C. Rosso, E. Vanni, E. Bugianesi, M. Cassader. Medical Sciences, University of Turin, Turin, Italy E-mail: [email protected] Background and Aims: In NAFLD subjects the flow of free fatty acids (FFA) towards liver is often increased due to insulin resistance, as well as de novo hepatic lipogenesis. There are few data on the relative importance of fasting vs postprandial lipid metabolism in NAFLD patients. We have studied the effect of a lipid load on FFA composition in 20 NAFLD subjects. The subjects were divided into 2 groups: mild fibrosis (n = 17) and severe fibrosis (n = 3). Methods: The patients were given a lipid load. Throughout the test we measured glucose, insulin, FFA composition, triglycerides


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POSTERS and cholesterol profile. FFA composition was performed with HPLC reversed-phase chromatography. Results: In NAFLD subjects with higher fibrosis the followings acids were significantly increased after a lipid load: Eicosapentaenoic acid was significantly increased at all times; Eicosatrienoic acid was significantly increased at times 0, 30, 60, 90, 120 minutes. Palmitoleic acids was significantly increased at times 30, 60, 90, 120, 150, 180 minutes. The area under the curve (AUC) of parameters measured during the oral fat test was computed by the trapezoid method. AUCs of eicosapentaenoic, eicosatrienoic and palmitoleic acids were significantly increased in NAFLD subjects with higher fibrosis score (all p < 0.002). Conclusions: Composition of FFA is different between subjects with mild vs severe fibrosis after a fat load. The different behavior of single FA observed in the postprandial phase could contribute to the development and progression of fatty liver disease. This is a pilot study; further investigations are needed to confirm these results. Funded by FP7/2007–2013 under grant agreement no. HEALTH-F2– 2009–241762 for the project FLIP and by PRIN 2009ARYX4T. P847 DAYTIME SLEEPINESS IN PATIENTS WITH NON-ALCOHOLIC FATTY LIVER DISEASE IS LINKED TO INSULIN RESISTANCE, ELEVATED LIVER FUNCTION TESTS AND FIBROSIS C. Bernsmeier1,2 , D.M. Weisskopf3 , J. Mosimann3 , B. Campana1,2 , A.C. Meyer-Gerspach2 , R.E. Steinert2 , L. Blaser2 , L. Jeker2 , L. Terracciano4 , C. Beglinger1,2 , M.H. Heim1,2 , C. Cajochen3 . 1 Division of Gastroenterology and Hepatology, 2 Department of Biomedicine, University Hospital Basel, 3 Centre for Chronobiology, Psychiatric Hospital of the University of Basel, 4 Institute of Pathology, University Hospital Basel, Basel, Switzerland E-mail: [email protected] Background and Aims: Sleep disruption is associated with the development of obesity and diabetes in murine models and humans. In Non-alcoholic fatty liver disease (NAFLD) fatigue has previously been associated with daytime sleepiness but not disease severity or insulin resistance (IR). Sleep apnoea is a risk factor for NAFLD. The role of sleep disturbance independent of its aetiology in the pathogenesis of NAFLD is unknown. We sought to detail sleep quality, daytime sleepiness, affect and clinical parameters in NAFLD patients. Methods: Patients with biopsy-proven NAFLD (n = 49) were compared to healthy controls (n = 25). Pittsburgh sleep quality index (PSQI), daytime sleepiness (Epworth sleepiness scale (ESS)), positive and negative affect scale (PANAS) were assessed and correlated to biochemical and histologic parameters. IR was quantified using HOMA-IR. Histology was evaluated using NAFLD activity score. Results: Sleep quality was poor in NAFLD compared to controls (PSQI 8.150 vs. 4.667, p = 0.0068) and associated with increased negative- and reduced positive affect. The ESS score overall was within normal range. However in NAFLD sleepiness was positively correlated to IR (HOMA-IR r = 0.33, p = 0.0344), liver function tests (LFTs) (AST r = 0.40, p = 0.0082, ALT r = 0.34, p = 0.0251) and fibrosis (r = 0.38, p = 0.0318). Positive affect itself was negatively correlated to LFTs (AST, r = −0.49, p = 0.0013, ALT r = −0.48, p = 0.0015) here. Conclusions: Daytime sleepiness in patients with NAFLD is linked to biochemical and histologic markers of disease activity. Poor sleep quality paralleled changes in affect. The data imply a pathogenic interaction of sleep disruption and NAFLD and might endorse studies exploring sleep regulation as a possible therapeutic tool.

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P848 SERUM CYTOKERATIN-18 IS ASSOCIATED WITH OXIDATIVE STRESS IN PATIENTS WITH NON-ALCOHOLIC FATTY LIVER DISEASE L. Polimeni, M. Del Ben, F. Baratta, M. Brancorsini, R. Carnevale, C. Nocella, C. Calabrese, D. Pastori, F. Angelico. Sapienza University of Rome, Rome, Italy E-mail: [email protected] Background and Aims: Non-alcoholic fatty liver disease (NAFLD) comprises a wide spectrum of liver damage ranging from simple steatosis to steatohepatitis, advanced fibrosis and cirrhosis. Hepatocyte apoptosis may play a major role in the progression of NAFLD. Cytokeratin-18 M30 fragment (CK-18), a marker of hepatocyte apoptosis, may predict severity of liver damage. Oxidative stress is considered one of the key mechanisms responsible for liver damage in NAFLD. The aim of the study was to determine the association between oxidative stress and CK-18 in a population of NAFLD patients. Methods: 209 patients with NAFLD were enrolled. Severity of steatosis was defined according to Hamaguchi’s ultrashonographic criteria. Subjects underwent routine clinical and biochemical evaluation. Serum levels of CK-18, urinary 8-iso-prostaglandin F2a (8-iso-PGF2a), soluble NOX2-derived peptide (sNOX2-dp) and adiponectin (APN) were measured. Results: Stratifying population by CK-18 tertiles, a significant progression of BMI, prevalence of severe NAFLD and diabetes, HOMA-IR, triglycerides and urinary 8-iso-PGF2a was found. Instead, HDL-cholesterol and APN progressively decreased. A positive correlation between CK-18 and BMI, HOMA-IR, Hamaguchi’s score, urinary 8-iso-PGF2a (r = 0.61; p < 0.001), sNOX2dp (r = 0.45; p < 0.001) and a negative correlation between CK-18 and HDLcholesterol and APN (r = −0.45; p < 0.001) were found. By stepwise multiple regression analysis, APN and sNOX2-dp were independent predictors of CK-18 levels. Conclusions: Patients with NAFLD and higher levels of serum CK-18 have a worse metabolic profile and increased oxidative stress. Levels of CK-18 are predicted by serum APN and sNOX2-dp. Our results suggest that oxidative stress may play a role in the promotion of apoptosis in patients with NAFLD. P849 QUANTITATIVE AND QUALITATIVE ANALYSIS OF PLASMA FREE FATTY ACIDS AND THEIR IMPACT ON LIVER DAMAGE IN NAFLD PATIENTS C. Saponaro1 , E. Vanni2 , M. Gaggini1 , C. Rosso2 , L. Mezzabotta2 , R. Gambino2 , E. Buzzigoli1 , F. Carli1 , M.L. Abate2 , F. Saba2 , F. Salomone3 , G.P. Caviglia2 , A. Smedile2 , M. Rizzetto2 , M. Cassader2 , E. Bugianesi2 , A. Gastaldelli1 . 1 Institute of Clinical Physiology, CNR, Pisa, 2 Dept. of Medical Sciences, Division of Gastroenterology and Hepatology and Lab. of Diabetology, University of Turin, Turin, 3 U.O.C. of Gastroenterology, Azienda Sanitaria Provinciale di Catania, Catania, Italy E-mail: [email protected] Background and Aims: The relative role of quality vs. quantity of fat delivered to the liver in the pathogenesis of NAFLD/NASH is currently unclear. The aim of this study was to investigate basal free fatty acids (FFA) flux and composition and their impact on the severity of liver damage in NAFLD patients. Methods: FFA composition was determined with Gas Chromatography Mass Spectrometry during fasting state in 54 subjects (45 patients with biopsy proven NAFLD, 9 healthy controls (CT)). De novo lipogenesis (DNL) index, stearoyl-CoA desaturase (SCD1) activity index, a desaturation index, and PUFA:SFA (P:S) ratio were derived from FFA composition. Adipose tissue insulin resistance (Adipo-IR) was calculated from Lipolysis*Insulin.


POSTERS Results: Total FFA were not different in NAFLD vs. CT (0.64±0.25 vs. 0.57±0.20). However, NAFLD patients with severe fibrosis (F3/F4) had higher levels of palmitoleic, palmitic and stearic acid (21.4±12.4 vs. 9.8±5.2; 171.3±68.7 vs. 115.4±45.0; 64.4±22.9 vs. 40.4±13.9, respectively; all p < 0.05). The medium P:S ratio, but not the short and long P:S ratios, was significantly decreased in F3/F4 patients (0.58±0.14 vs 0.87±0.11, p < 0.001) and was negatively correlated to the increase in the Adipo-IR (r = 0.40). DNL and SCD1 indexes were significantly increased in NAFLD compared to CT, particularly in those with severe fibrosis (ANOVA p < 0.008 and p < 0.002, respectively), and correlated positively with lipolysis (r = 0.43 and r = 0.40) and Adipo-IR (r = 0.49 and r = 0.30 respectively).

Figure: DNL index and SCD-1 index in CT and NAFLD patients with different degrees of fibrosis. *p < 0.05 vs. CT; § p < 0.002 vs. CT.

Conclusions: Both quantitative and qualitative alterations in lipid metabolism contribute to the progression of liver damage in NAFLD patients. Funded by FP7/2007–2013 under grant agreement no. HEALTH-F2– 2009–241762 for the project FLIP and by PRIN 2009ARYX4T. P850 PERIPHERAL AND HEPATIC VEIN CYTOKINE LEVELS IN CORRELATION WITH NON ALCOHOLIC FATTY LIVER DISEASE (NAFLD) METABOLIC, HISTOLOGICAL, AND HEMODYNAMIC FEATURES L. Vonghia1,2 , T. Magrone2 , A. Verrijken3 , P. Pelckmans1,4 , P. Michielsen1,4 , L. Van Gaal3 , E. Jirillo2 , S. Francque1,4 . 1 Gastroenterology and Hepatology, Antwerp University Hospital, Antwerp, Belgium; 2 Basic Medical Science, University of Bari, Bari, Italy; 3 Endocrinology, Diabetology and Metabolism, 4 University of Antwerp, Antwerp, Belgium E-mail: [email protected] Background and Aims: Haemodynamic impairment, inflammatory mediators and glucose metabolism impairment have been implicated in the pathogenesis of Non Alcoholic Fatty Liver Disease (NAFLD). The aim is to study the cytokine profile in patients with NAFLD/Non Alcoholic Steatohepatitis (NASH) in peripheral (P) and hepatic vein (HV) blood and to compare with histology, haemodynamic and metabolic parameters. Methods: 40 NAFLD/NASH obese patients who underwent a transjugular liver biopsy were enrolled. Besides an extended liver and metabolic work-up, IL1B, IL4, IL6, IL10, IL23, TNFa and INFg were measured in plasma obtained from P and HV blood by means of multiplex immunoassay. The Th1/Th2 (INFg/IL4), the M1/M2 [(TNFa+IL6+IL23)/IL10] and the IL10/IL17 ratios were calculated. Results: A decrease of the P-IL10/IL17-ratio and an increase of the P-M1/M2-ratio (p < 0.05) was observed in NASH versus NAFLD patients. A P-M1/M2-ratio increase was detected also in patients with portal hypertension in comparison with patients without it (p < 0.05). Moreover diabetic patients showed an increase of the P-Th1/Th2-ratio in comparison with non diabetic ones (p < 0.05). The P-M1/M2 ratio positively correlated with steatosis grade (r:0.39,p:0.02) and insulin (r:0.47,p:0.003). The HV-M1/M2 ratio positively correlated with fasting insulin and Hepatic Vein Pressure Gradient (r:0.47,p0.003). IL6 correlated with the visceral fat amount (r:0.36,p:0.02). The P- and HV-IL10/IL17 ratios negatively correlated with fasting insulin (respectively r:-0.4,p:0.005;r:0.4,p:0.01).

Conclusions: A proinflammatory cytokine state is associated with more disturbed metabolic, histological, and hemodynamic features in NAFLD obese patients. An increase of the M1/M2 ratio and a decrease of the IL10/IL17 ratio play a key role in this process. P851 PREDIABETES CONTRIBUTES TO THE DEVELOPMENT OF FATTY LIVER: A LONGITUDINAL STUDY A. Moriya1 , Y. Iwasaki2 , S. Ohguchi3 , E. Kayashima3 , T. Mitsumune3 , H. Taniguchi4 , M. Ando1 . 1 Department of Medicine, Mitoyo General Hospital, Kanonji, 2 Health Service Center, Okayama University, 3 Junpukai Health Maintenance Center, Okayama, 4 Department of Medicine, Tottori Municipal Hospital, Tottori, Japan E-mail: [email protected] Background and Aims: Type 2 diabetes is closely associated with fatty liver. Impaired fasting glycaemia (IFG) is also supposed to be associated with the incidence of fatty liver. Methods: We obtained clinical and laboratory data from Japanese subjects who voluntarily underwent a baseline health checkup and follow-up studies at least once from 2006 to 2011. We excluded cases with concurrent liver diseases or taking medication for diabetes and included 5,954 cases without fatty liver (3,058 men and 2,896 women; median age, 45.1 years; median BMI, 21.6 kg/m2 ). We stratified participants into following 4 groups according to fasting plasma glucose (FPG) level: 125 mg/dL. The presence of fatty liver was assessed by ultrasonography. Using Cox proportional hazard model, we estimated the influence of IFG on the incidence of fatty liver. Results: During the total follow-up period of 12,164 personyears, 703 incidences of fatty liver were observed. Each risk ratio (95% confidence intervals) for FPG 100–109 mg/dL, 110–125 mg/dL, and >125 mg/dL was 1.98 (1.68–2.33), 2.03 (1.55–2.67), and 3.65 (2.36–5.66), respectively. After adjustment for age, sex, and BMI, only FPG 100–109 mg/dL remained significant (P < 0.001). Conclusions: FPG level from 100 to 109 mg/dL was associated with the incidence of fatty liver. On the other hand, BMI is supposed to be a dominant risk factor in cases with severer IFG. P852 WHY GGT LEVELS SHOULDN’T BE DISREGARDED WHEN CONSIDERING A PATIENT WITH POSSIBLE NAFLD N. Barsic1 , D. Radic2 , Z. Marusic3 , M. Gomercic Palcic1 , S. Stojsavljevic1 , M. Duvnjak1 . 1 Dept. of Gastroenterology and Hepatology, Zagreb University School of Medicine, Sestre Milosrdnice University Hospital Center, 2 Dept. of Gastroenterology and Hepatology, Zagreb University School of Medicine, University Hospital Center Zagreb, 3 Dept. of Pathology, Zagreb University School of Medicine, Sestre Milosrdnice University Hospital Center, Zagreb, Croatia E-mail: [email protected] Background and Aims: In clinical practice and research related to NAFLD, GGT levels are often disregarded, although no evidence justifies that. As there are no data on NAFLD characteristics in patients with isolated GGT elevation, we aimed to analyse this subgroup and compare it with patients with elevated ALT/AST. Additionally, we investigated the clinical observation that GGT elevation is much more frequent than ALT/AST in patients with NAFLD features, as data on relative prevalence of liver enzyme elevation in NAFLD are very scarce. Methods: Detailed analysis was peformed in 84 prospectively included patients with biopsy-confirmed NAFLD, including 24 patients with repeatedly isolated GGT elevation. Diabetes clinic’s electronic chart database was utilised to analyse liver enzyme levels and other data in 913 type 2 diabetes patients, including 156 patients with BMI >35 (who should virtually all have NAFLD). Results: Comparing patients with isolated GGT elevation and those with ALT elevation, there were no significant differences


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POSTERS in severity of the pathohistological features, including Brunt necroinflammatory degree (severe degree 26.3% vs. 18.3%) and NAFLD Activity Score (median 5.0 vs. 4.5), except slightly in fibrosis stage (advanced fibrosis 12.5 vs. 21.7%). In analysis of overweight type 2 diabetic population, prevalence of elevated GGT vs. ALT was 49.5% vs. 26.0% in BMI >35 group, and 30.0% vs. 15.2% in lower BMI groups. Conclusions: NAFLD patients with isolated GGT elevation demonstrate similar pathohistological disease severity as those with elevated ALT, and should be approached for liver biopsy and pharmacological therapy in the same way. GGT elevation is 2x more frequent than ALT elevation. P853 BERBERINE WITH ALFA LIPOIC ACID (ALA) IN NON ALCOHOLIC STEATO-HEPATITIS (NASH). A RANDOMIZED DOUBLE BLINDED PLACEBO CONTROL TRIAL. A CLINICAL PILOT – THE BANISH TRIAL P.P. Basu1,2 , N.J. Shah3 , M. Aloysius2 , R.S. Brown Jr1 . 1 College of Physicians and Surgeons, Columbia University, 2 King’s County Hospital Medical Center, New York, 3 James J. Peters VA Medical Center, Icahn School of Medicine at Mount Sinai, Bronx, NY, United States E-mail: [email protected] Background and Aims: Therapeutic modalities for NASH have not yet been fully established. Anti-oxidants and insulin sensitizers altering insulin resistance, inhibiting intra hepatic oxidative stress with inhibition of formation of free radicals and blocking inflammatory cytokines to prevent fibrosis. Berberine is a natural substance extracted from plants like Berberis which is found to up-regulate intra hepatic pathways as insulin sensitizer, via GLP-1 up regulation and Acyl palmotyl mechanism on fatty acid oxidation, induction of PPAR gama; all of which blocks the terminal inflammatory Cytokine release TNF Alfa to prevent fibrosis that prevents End stage liver disease (ESLD) and liver cancer. Methods: Hundred and twenty patients (n = 120) with NASH were recruited. Mean BMI 29.9% (29% to 32%) with 69 males and 51 females. Hispanic 46, Caucasians 34, Asian Pacific15, Black 11, Asian 14. Mean HbA1C 6.2 (5.9–6.8), Mean HOMA 2.7 (2.1–3.6), ALT 54 (38–79), Triglyceride 287 (233–344), LDL c 163 (129–176), Leptin 63 (43–98), Adiponectin 0.9 (0.1–1.1), RBP 4 of 5.8 (4.0–6.8), TNF Alfa 3.8 (2.1–4.8), IL 12 of 5.3 (3.9–7.8), Serum Fibrotic and Steatotic scores were measured at 0 and then at 6 months. Results: See the table.

Group Group Group Group

A B C D

ALT

HOMA TG

Adiponectin TNF-a

Leptin Steatotic score

0.001 60%: 100%, 57% and 24%, respectively; Fibrotest AF (score ≥0.48) 12 pts (29%); Steatotest presumed steatosis >5% and >30% in 96% and 81%, respectively. The median (range) FU was 2.3 yrs (1.0–7.3). Evolution of steatosis: N = 9 pts LB progression [mean(se) Steatotest evolution: 0.70(0.04) vs 0.76(0.03), p = NS]; N = 8 pts stable LB steatosis [Steatotest 0.66(0.08) vs 0.68(0.08), p = NS]; N = 25 pts had LB regression [Steatotest 0.68(0.03) vs 0.61(0.03), p = 0.03]. During follow-up 7/42 pts progressed fibrosis stage classification on LB. According to Steatotest [follow-up = 2.5(0.3) yrs] the HR (95% CI) to progress at least one fibrosis histological stage (SAF-fibrosis score) was 24.97 (0.03–20,899) in subjects with Nashtest scores predicting NASH (= 0.75) versus 0.04 (0.00–33.52) in subjects with Nashtest scores predicting no NASH/borderline NASH (= 0.25/0.50). The same figures were obtained using the fibrosis progression estimated by Fibrotest (progression of 0.10): HR = 2.8 (0.0–7377.9) vs 0.4 (0.0–969.9). Conclusions: This study confirmed the accuracy of Steatotest to reflect dynamic histological changes in NAFLD-patients. In subjects with Steatotest progression, hazard ratio to develop fibrosis was higher in subjects with Nashtest predicting NAS (Kleiner-score, Hepatology 2005).

P862 LONGITUDINAL VALIDATION OF BIOMARKERS OF FIBROMAX PANEL, AS SURROGATES FOR DYNAMIC HISTOLOGICAL CHANGES ON LIVER BIOPSY (LB) IN NAFLD PATIENTS M. Munteanu1 , R. Pais2 , Y. Ngo1 , F. Charlotte3 , L. Fedchuk4 , P. Lebray2 , P. Bedossa5 , V. Ratziu2 , T. Poynard2 , LIDO Study Group. 1 Hepatology Research Unit, BioPredictive, 2 Department of Hepatology GH Pitié-Salpêtrière, 3 Pathology Department, APHP UPMC Paris Liver Center, 4 ANRS, Paris, 5 Pathology Department, CHU Beaujon, Clichy, France E-mail: [email protected] Background and Aims: No longitudinal studies used repeated LB to evaluate surrogate markers surrogate-markers (FibroMax™) during longitudinal follow-up (FU) in NAFLD-patients (PlosOne 2010). Aim: Evaluate in NAFLD-patients: 1. Dynamics of liver estimate Steatotest according to LB; 2. The impact on fibrosis of presumed steatosis (Steatotest) and NAS (Nashtest) using two repeated FibroMax and histological fibrosis. Methods: Patients from previously published study (J Hepatol 2013) with persistent-NAFLD, selected with 2-repeated LB and FibroMax (panel including Fibrotest/Steatotest/Nashtest) were Journal of Hepatology 2014 vol. 60 | S215–S359

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Poster Session – Saturday, 12 April 2014

1B. LIVER TRANSPLANTATION/SURGERY: CLINICAL

P863 A SIMPLE MODEL TO PREDICT 5-YEAR MORTALITY AFTER LIVER TRANSPLANTATION IN PATIENTS WITH END-STAGE LIVER DISEASE F.S. Cardoso1 , S.M. Bagshaw1 , J.G. Abraldes2 , N. Kneteman3 , G. Meeberg4 , P. Fidalgo1 , C.J. Karvellas1,2 . 1 Division of Critical Care, 2 Division of Gastroenterology, 3 Department of Surgery, 4 Transplantation, University of Alberta, Edmonton, AB, Canada E-mail: [email protected] Background and Aims: A simple tool for estimating the long-term survival of patients with end-stage liver disease after orthotopic liver transplantation (OLT) would optimize decisions on eligibility for OLT made by clinicians. Firstly, we aimed to externally validate the previously derived Charlson Comorbity Index for OLT (CCIOLT). Secondly, we aimed to develop a new model to predict 5-year mortality after OLT. Methods: This single center retrospective cohort study included 524 consecutive adult cirrhotic patients who underwent OLT between 2002 and 2012. External validation of CCI-OLT used the Kaplan–Meier method. Derivation of the new predictive model used Cox proportional hazards regression.

Figure: Nomogram for prognosis of adult patients with end-stage liver disease after OLT.

OLT based on six easy-to-know pre-transplant characteristics of patients: age, body mass index, hepatitis C, hepatic encephalopathy, ICU stay at transplant, and live donor transplant. Based on this predictive model, we further developed a nomogram to allow clinicians to easily estimate individual probability of 1-, 3-, and 5-year survival after OLT. Conclusions: A new model to assess prognosis of adult patients with end-stage liver disease after OLT was derived. This is a simple rule to help clinicians quantify the individual expected survival benefit with OLT. P864 INFERIOR OUTCOME OF LIVER TRANSPLANTATION FOR PRIMARY SCLEROSING CHOLANGITIS IN THE MODEL FOR END-STAGE LIVER DISEASE ERA J. Klose1,2 , M. Klose2 , M. Manns2 , J. Klempnauer2 , H. Schrem2 . 1 University Hospital Heidelberg, Heidelberg, 2 Hannover Medical School, Hannover, Germany E-mail: [email protected] Background and Aims: Survival after liver transplantation (LTX) has decreased in Germany since the implementation of MELD-based liver allocation. Primary sclerosing cholangitis (PSC) is regarded to be one of the best indications for LTX. However, even for PSC the outcome after LTX is supposed to be deteriorated. The purpose was to investigate the results after LTX for PSC at our center. Methods: The influence of MELD-based liver allocation on the outcome of LTX for PSC is analyzed in this retrospective study, including 126 consecutive patients treated with LTX for PSC (01.01.1999–31.08.2012; pre-MELD-era 01.01.1999–31.12.2006: n = 85, MELD-era 01.01.2007–31.08.2012: n = 41). Results: Survival following LTX was significantly higher in the preMELD-era (p = 0.008; Log Rank). Mean waiting time was longer for patients in the MELD-era (2.3 years versus 1.6 years; p = 0.365, Chi2 ). Although patients in the MELD-era had a higher incidence of dominant bile duct stenosis (58.5% versus 35.3%; p = 0.048; Chi2 ) there was no higher frequency of cholangiocarcinoma prior to LTX (4.9% versus 8.2%, p = 0.493, Chi2 ) or in explanted livers (7.3% versus 7.1%, p = 0.958). Patients within the MELD-era who gained exception points during their waiting time had a longer overall waiting time (mean 2.67 (n= 26) versus 1.64 years (n = 15), p = 0.045, Mann–Whitney-U-Test) while waiting time from the first award of exception points was clearly reduced compared to cases without any exceptions points until LTX (mean 191 days versus 567 days, p = 0.383, Chi2 ). Conclusions: Current liver allocation for PSC-patients should be reconsidered. Earlier transplantation could improve the survival rate following LTX for PSC-patients.

Results: The 1-, 3-, and 5-year cumulative survival after OLT was 89%, 80%, and 73%, respectively. CCI-OLT was not associated with 5-year mortality after OLT (P = 0.34). We derived and internally validated a new predictive model of 5-year mortality after

Journal of Hepatology 2014 vol. 60 | S361–S522 © 2014 All rights reserved.

POSTERS P865 IS HEPATIC RESECTION INDICATED ONLY FOR SINGLE SMALL HEPATOCELLULAR CARCINOMA? VALIDATION OF THE EASL/AASLD GUIDELINES BASED ON OUR EXPERIENCE S. Tanaka1 , Y. Iimuro1 , T. Hirano1 , S. Hai1 , I. Nakamura1 , Y. Kondo1 , K. Suzumura1 , T. Okada1 , S. Nishiguchi2 , J. Fujimoto1 . 1 Department of Surgery, 2 Division of Hepatobiliary and Pancreatic Diseases, Department of Internal Medicine, Hyogo College of Medicine, Nishinomiya, Japan E-mail: [email protected] Background and Aims: The EASL/AASLD guidelines, based on the BCLC classification, recommend hepatic resection only for patients with single small (≤ 5 cm) hepatocellular carcinoma (HCC) without portal hypertension, hyperbilirubinemia or macrovascular invasion. This study investigated which patients benefit from hepatic resection. Methods: The clinical records of 566 patients who underwent initial resection for HCC were retrospectively reviewed. According to the BCLC classification, 79 patients (14%) had BCLC 0, 284 (50%) BCLC A, 148 (26%) BCLC B and 55 (10%) had BCLC C. An analysis of the overall survival (OS) and a multivariate analysis of prognostic factors were performed. Results: The in-hospital mortality rate was 2.1% (12/566). The three- and five-year OS rates were 92% and 84% for BCLC 0; 76% and 60% for BCLC A, 72% and 54% for BCLC B and 25% and 13% for BCLC C (p < 0.0001). Among the 363 patients with BCLC 0-A, 262 patients fit the guidelines for hepatic resection (five-year OS rate; 66%), 14 were recommended to undergo transplantation (five-year OS rate; 100%) and the remaining 87 were recommended to undergo ablation (fiveyear OS rate; 59%, p = 0.28). The multivariate analysis identified low albuminemia, cirrhosis, a-fetoprotein ≥ 20 ng/mL, multiple, macrovascular invasion, and intraoperative blood loss ≥ 1 L as independent prognostic factors for the OS, but hyperbilirubinemia, portal hypertension and large tumor (>5 cm) were not. Conclusions: The outcomes after hepatic resection for HCC are acceptable except for BCLC C patients. Hepatic resection may be useful for some patients with BCLC 0, A and B. P866 EVEROLIMUS-BASED IMMUNOSUPPRESSION PROVIDES SUPERIOR RENAL FUNCTION AND COMPARABLE EFFICACY VERSUS STANDARD TACROLIMUS IN DE NOVO LIVER TRANSPLANTATION: 3-YEAR RESULTS FROM H2304 STUDY EXTENSION F. Saliba1 , G.M. Kaiser2 , L. De Carlis3 , H.J. Metselaar4 , P. De Simone5 , C. Duvoux6 , F. Nevens7 , L. Fischer8 , J. Fung9 , G. Dong10 , B. Rauer11 , opital Paul Brousse, Villejuif, France; 2 University Hospital G. Junge11 . 1 Hˆ a Granda’ Hospital, Milan, Italy; Essen, Essen, Germany; 3 Niguarda ‘C` 4 Erasmus MC, University Hospital Rotterdam, Rotterdam, Netherlands; 5 Azienda Ospedaliero-Universitaria Pisana, Pisa, Italy; 6 Hˆ opital Henri Mondor, Creteil, France; 7 University Hospital Gasthuisberg, KU Leuven, Belgium; 8 University Medical Center Hamburg-Eppendorf, Hamburg, Germany; 9 Cleveland Clinic Foundation, Cleveland, OH, 10 Novartis Pharmaceuticals Corporation, East Hanover, NJ, United States; 11 Novartis Pharma AG, Basel, Switzerland E-mail: [email protected] Background and Aims: Exposure to calcineurin inhibitors (CNI) is a well-recognized risk factor for progressive kidney disease after liver transplantation (LTx). The H2304 Extension (E1) study provides prospectively collected 3-year data on renal function, efficacy and safety with everolimus (EVR) and reduced-exposure to tacrolimus (rTAC) versus standard TAC (TAC-C). Methods: In H2304E1, 282 patients (of 719 patients in core study) continued on EVR+rTAC (EVR C0 3–8 ng/mL+ rTAC C0 3–5 ng/mL; N = 106); TAC-WD (EVR C0 6–10 ng/mL with TAC withdrawal at M4; N = 51); or TAC-C (C0 6–10 ng/mL; N = 125). Composite efficacy S362

failure rate (CEF; treated biopsy proven acute rejection [tBPAR], graft loss or death), renal function (eGFR by MDRD4), and safety were evaluated at Month (M) 36. Results: From randomization to M36, CEF rate was comparable between EVR+rTAC and TAC-C arms (11.5% and 14.6%, p = 0.334). Incidence of BPAR was lower in EVR+rTAC arm (7.3%) versus TAC-C (17.7%; p = 0.006). Superior renal function was maintained up to M36 in EVR+rTAC versus TAC-C arm (mean eGFR: 78.7 versus 63.5 mL/min/1.73m2 ; p < 0.001) (Figure). During the extension phase, EVR+rTAC and TAC-C arms exhibited comparable incidence of adverse events (AEs; 82.1% versus 76.8%), serious AEs (30.2% versus 22.4%) and discontinuation of study medication due to AEs (8.5% versus 7.2%). At M36, in TAC-WD arm mean eGFR was 85.5 mL/min/1.73m2 , CEF was 27.4% (1 tBPAR during extension) and incidence of AE was 94.1%. Conclusions: EVR+rTAC regimen started 1 month post-LTx, provides superior renal function sustained for 3 years with lower BPAR rates and overall comparable efficacy and safety versus TAC-based regimen.

Figure: Mean eGFR (MDRD4) (all extension patients).

P867 FACTORS PREDICTING OUTCOME OF PATIENTS WITH ACUTE LIVER FAILURE MEETING THE CRITERIA OF LIVER TRANSPLANTATION: IMPACT OF ALBUMIN DIALYSIS WITH MARS F. Saliba1,2,3 , G. Dahlqvist1 , A. Letierce4 , P. Ichai1,2,3 , I. Ruiz1 , opital M. Boudon1 , D. Samuel1,2,3 . 1 Centre Hepato-Biliaire, AP-HP, Hˆ Paul Brousse, 2 Univ Paris-Sud, 3 Unit 785, Inserm, Villejuif, 4 Centre de Recherche Clinique, AP-HP, Hˆ opital Bicêtre, Le Kremlin-Bicêtre, France E-mail: [email protected] Background and Aims: Despite recent improvement in the outcome of patients with acute liver failure (ALF) undergoing liver transplantation (LT), the overall mortality taking into account all patients meeting the criteria of LT admitted to a liver unit remains high. Methods: A single centre retrospective analysis of all patients who were admitted to the liver ICU for ALF and met the criteria of LT. Results: From 2008 till 2012, 72 patients (61.1% female) with a mean age of 45.1+16.1 years were admitted for acute liver failure and met either the Clichy-Paul Brousse criteria (70.8%) or the King’s College criteria (77.8%) or both criteria (62.5%) for LT. Paracetamol etiology was present in 29 patients (40.3%). Hepatic encephalopathy was present at admission in 73.6% of the patients or developed early within 24–48 hours. 42 patients (58.3%) required mechanical ventilation. Mean MELD and SOFA scores were respectively 35.5+5.9


POSTERS and 8.4+4.3. Forty-six patients (63.9%) received albumin dialysis treatment with MARS® with a median duration of 14.5 hours (range: 3–56). Only 32 patients (44.4%) underwent a liver transplantation. Survival was observed in 77.8% of the patients: 87.5% in transplanted and 70% in non-transplanted. The main marker differentiating those patients transplanted from those non-transplanted was the lactate level which was significantly higher in transplanted patients (8.2+7.9 vs. 4.5+3.8 mmol/L; p = 0.03). Conclusions: High lactate level at admission was the main predictive factor of severity of ALF and the need for LT. Optimal medical management combining MARS® treatment reduced the need for LT with an optimal survival. P868 CHANGE IN MELD AT 2 WEEKS AFTER ACUTE-ON-CHRONIC LIVER FAILURE AS INDICATOR OF MORTALITY AND LIVER TRANSPLANT AT 60 DAYS R. Kumar, T.L. Krishnamoorthy, H.K. Tan, H.F. Lui, W.C. Chow. Department of Gastroenterology and Hepatology, Singapore General Hospital, Singapore, Singapore E-mail: [email protected] Background and Aims: Acute on chronic liver failure (ACLF) is characterised by sudden deterioration of underlying chronic liver disease resulting in increased mortality and need for liver transplant. Identifying the patients at risk of increased mortality is essential for early intervention and prognostication. Methods: ACLF was defined as per Asiapacific association of liver disease (APASL) criteria. Inpatient discharge summaries of patients admitted between January 2001 to April 2013 were scrutinized to identify patients fulfilling the above criteria. The primary outcome measured was mortality or liver transplant within 60 days from the onset of ACLF. Change in MELD score at 1, 2 and 4 weeks was assessed to predict mortality or liver transplant. Results: 55 subjects fulfilled the above criteria, data was available for 53 subjects at time of reporting. The mean age of patients was 55.9±10.6 years with 73.6% male. The most common cause for ACLF was Hepatitis B flare (52.8%), Alcohol (18.9%) and drugs (9.4%). Mortality at 60 days was 36% while 4 (7.5%) had liver transplant done. Mean survival duration for patients who died was 27±17 days. Improvement of MELD score ≥0 at 2 weeks had negative predictive value of 93.75%, while worsening of MELD score ≥3 at 2 weeks had positive predictive of 77.27% for mortality or liver transplant. Conclusions: Change of MELD score at 2 weeks provides opportunity for early prognostication. Improvement in MELD score has negative predictive value while worsening of more than 3 has positive predictive value for mortality or liver transplant. This data needs further validation. P869 A COMPARATIVE STUDY BETWEEN LARGE PAPILLARY BALLOON DILATATION AND MECHANICAL LITHOTRIPSY FOR THE MANAGEMENT OF DIFFICULT CBD STONES M. Naga1 , A. ElBadri1 , S. Elkholy1 , S. Nabil2 . 1 Internal Medicine, Faculty of Medicine – Cairo University, 2 Helwan, University Hospital, Cairo, Egypt E-mail: [email protected] Background and Aims: Extraction of difficult bile duct stones may require mechanical lithotripsy (ML) as an adjunctive procedure, which lengthens the procedure time. Recent studies have shown that LBD is safe and effective in these patients. Aim: To assess effectivness of new techniques (LBD) in managing difficult CBD stoness. Methods: This study was conducted on 100 patients with calcular obstructive jaundice with difficult stone extraction done by single endoscopist, they were classified into 3 groups the 1st group

included patients in which ML was used 2nd group LBD was used 3rd group both techniques were used. Then success rate was determined with a final cholangiogram, whereas type and rate of complications were assessed. Results: The highest success rate was in the 2nd group 93.8% followed by the 3rd group 90% then 1st group 87%. 22% of the patients in the 1st group needed repeated sessions while in the 2nd and 3rd groups its 6.2% and 10% respectively. Bleeding was higher in the 2nd and 3rd group than the 1st one. No perforation was done, 2 cases showed minor tear during dilatation for which the procedure was stopped. Conclusions: LBD could be superior to ML in patients with difficult stone extraction regarding the success rate, number and time of sessions. In cases as Mirrizzi syndrome (type 3) and liver cirrhosis ML is preferred. In cases of associated tight strictures combined techniques are preferred. Bleeding was higher in LBD. Position of cystic duct above the upper tip of the balloon is essential to avoid its tear. P870 REVERSIBLE NON-ISCHEMIC CARDIOMYOPATHY WITH SEVERE LEFT VENTRICULAR DYSFUNCTION AFTER LIVER TRANSPLANTATION M. Yataco1 , T. Difato1 , J. Trejo-Gutierrez2 , T. Patel1 , B. Rosser1 , J. Bargehr1 , S. Pungpapong1 , B. Taner1 , J. Aranda-Michel2,3 . 1 Transplantation, 2 Mayo Clinic, Jacksonville, FL, 3 Liver Transplantation, Swedish Health Systems, Seattle, WA, United States E-mail: [email protected] Background and Aims: New onset non-ischemic cardiomyopathy (NIC) has been reported after liver transplantation (LT). Our aims were to define the prevalence, associated clinical factors and progression of NIC. Methods: Medical records of all LTs done at our institution between 2005–2012 were reviewed and patients who developed NIC were identified. Results: Seventeen recipients out of 1460 LTs (1.2%) developed NIC. Median pre-LT QTc interval was 459 (range, 405–530). Median pre-LT ejection fraction was 65% (range, 50–81) and median E/A ratio was 1 (range, 0.71–1.67). Prior to LT, nine patients (53%) were admitted to ICU, fourteen patients (82%) were severely malnourished, thirteen patients (76%) had renal insufficiency and eleven patients (65%) required CRRT. Median raw MELD score was 29 in patients with NIC versus 18 in no-NIC patients (p = 0.01). There was no significant difference between NIC and no-NIC patients regarding donor age, donor risk index, cold ischemia and warm ischemia time. Median time of onset was 2 days post LT (0–20 days) and diagnostic echocardiograms showed a median ejection fraction 21% (15–32%). One patient died two months post-LT. Recovery of cardiac function occurred in 16 patients, with a median ejection fraction of 44% (25–65%) at the time of discharge. Median length of hospitalization was 26 days (9–63 days) in patients with NIC versus 8 days (0–515 days) in no-NIC patients (p = 0.01). One year survival of NIC patients was 94.1%. Conclusions: Patients who developed NIC are frequently critically ill with a high MELD score, renal insufficiency and severe malnutrition. P871 SIGNIFICANTLY INCREASED POST-LIVER TRANSPLANTATION MORTALITY AMONG PATIENTS AGE 70 YEARS AND OLDER R.J. Wong1 , A. Ahmed1 , C. Esquivel2 . 1 Gastroenterology and Hepatology, 2 Surgery, Stanford University School of Medicine, Stanford, CA, United States E-mail: [email protected] Background and Aims: Older age at time of liver transplantation (LT) is associated with higher risk of peri-operative complications. However, the impact of age on long term post-LT survival is not


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POSTERS clear. We evaluated the impact of age at time of LT on long term post-LT survival among U.S. adults. Methods: Using data from the United Network for Organ Sharing registry, we retrospectively evaluated adult LT recipients from 2002–2012. Age at transplantation was categorized into four groups (35 (vs. BMI 40 (vs. BMI 400 (p < 0.001). Normalization of the IBI post TACE was associated with radiologic response (p < 0.001) (Figure 1B) and improved OS and remained as significant multivariate predictor of OS in both the derivation and validation set (p < 0.001). Conclusions: Normalization of IBI after TACE is shown to be an independent predictor of survival and may integrate the retreatment criteria for repeat TACE in intermediate stage HCC. IBI and its dynamic changes after treatment are validated stratifying biomarker allowing for the identification of patients with a significant survival advantage following initial TACE. P976 RETREATMENT WITH TRANSARTERIAL CHEMOEMBOLIZATION (TACE): THE ABCR SCORE, AN AID TO THE DECISION-MAKING PROCESS X. Adhoute1 , G. Penaranda2 , J.-L. Raoul3 , J.-P. Bronowicki4 , P. Castellani1 , H. Perrier1 , O. Monnet1 , O. Bayle1 , P. Beaurain1 , B. Pol1 , C. Bazin4 , S. Naude4 , V. Oules1 , G. Lefolgoc1 , M. Bourliere1 . 1 Hˆ opital Saint Joseph, 2 AlphaBio Laboratory, 3 Instiut Paoli Calmettes, Marseille, 4 CHU Brabois, Nancy, France E-mail: [email protected]

(A)

(B)

Background and Aims: TACE is the standard of care for intermediate stage hepatocellular carcinoma (HCC). However, side effects are frequent and can impair survival. The ART score (based on the increase in AST, Child–Pugh score and the absence of radiologic tumour response) identifies patients who should not continue with TACE after one session. Sieghart W Hepatology 2013. Methods: From 01/2007 to 04/2012, 139 consecutive patients, mostly with viral-induced disease, were treated for HCC by TACE. We used the ART score to determine its prognostic value. Using the same methodology that built the ART score we calculated a new score in our population and we validated it in two cohorts of patients (internal and external). Results: The ART score with a cut-off value at 2.5 differentiated two groups with different survival, but the prognosis was not linearly correlated with the score. In a multivariate analysis we found four prognostic factors associated with OS: baseline BCLC and AFP, increase in Child–Pugh score, absence of radiological response. We calculated a new score (ABCR score) that was correlated with survival. This score was validated in an internal cohort of 53 patients and in an external cohort of 100 patients treated in University Hospital of Nancy. Increase in ABCR score was correlated with the prognosis, unlike ART score. Conclusions: The ART score was not of major value in our HCC population with mainly viral disease. The ABCR score had an excellent value and was well correlated with the prognosis. These results should be confirmed in a prospective study.

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POSTERS P977 UNRESECTABLE HEPATOCELLULAR CARCINOMA (HCC) TREATED BY TRANSARTERIAL CHEMOEMBOLIZATION (TACE): AFTER TWO SESSIONS, ART OR ABCR SCORES TO GUIDE THE DECISION MAKING PROCESS? X. Adhoute1 , G. Penaranda2 , J.-L. Raoul3 , P. Castellani1 , H. Perrier1 , O. Monnet1 , C. Muller1 , O. Bayle1 , M. Campanile1 , G. Lefolgoc1 , opital Saint Joseph, 2 AlphaBio S. Benaly1 , V. Oules1 , M. Bourliere1 . 1 Hˆ Laboratory, 3 Institut Paoli Calmettes, Marseille, France E-mail: [email protected] Background and Aims: TACE is the standard of care for intermediate stage HCC, for more advanced HCC in Asian recommendations. There is no consensus about the modalities of TACE. The ABCR score computed before the second TACE, is based on the baseline rate of AFP, BCLC stage, worsening Child– Pugh score, radiological response. It differentiated 3 groups with different overall survival and can guide treatment decisions. It is better correlated with the prognosis than the ART score. Aim: To evaluate and compare ART and ABCR scores after the second sessions of TACE. Methods: This single-center retrospective study includes 126 patients with unresectable BCLC B (45%), BCLC A (45%) and BCLC C HCC (10%) with segmental portal vein thrombosis, treated from 01/2007 to 06/2012 by at least 3 TACE, without further treatment, with Child–Pugh A or B cirrhosis. Results: Patients were treated on average by 4 sessions of TACE. The mean time to radiographic progression was 6 months [5.5–8]. The median follow-up was 21.8 months [17.3–26.4]. The median survival of all the patients was 28.4 months [23.5–33.4]. ART score differenciated two groups with significantly different survival. But increase in the score was not correlated with prognosis. The ABCR score differenciated three groups with different survival and increase in the score was better correlated with prognosis than the score ART (linear correlation coefficient R2 = 0.94 vs 0.21 (ART), p < 0.0001). Conclusions: The ABCR score is still an aid to the decision making processes after the second TACE. It is better correlated with survival than ART score. P978 ESTABLISH A RISK SCORE SYSTEM COMPOSED OF SEROLOGICAL, PATHOLOGICAL AND TUMOR BIOLOGICAL PARAMETERS TO PREDICT POSTOPERATIVE HEPATOCELLULAR CARCINOMA RECURRENCE J.-C. Wu1,2 , C.-W. Su1,3 , G.-Y. Chau4,5 . 1 Institute of Clinical Medicine, National Yang-Ming University, 2 Department of Medical Research, 3 Division of Gastroenterology, Department of Medicine, 4 Division of General Surgery, Department of Surgery, Taipei Veterans General Hospital, 5 Department of Surgery, National Yang-Ming University, Taipei, Taiwan E-mail: [email protected] Background and Aims: Postoperative recurrence is the major mortality cause for hepatocellular carcinoma (HCC) patients after surgery. In this study, we integrated serological, pathological and tumor biological parameters to establish a risk score system to predict HCC recurrence. Methods: One hundred and twenty-three HCC patients underwent surgery in Taipei Veterans General Hospital between 1990 and 2003 were included as a training set. A risk score system to predict post-operative recurrence weighted by b-coefficients in multivariate analysis of the training set was validated in another cohort of 162 HCC patients. The expression of E-cadherin was evaluated by immuno-histochemical staining. Results: By multivariate analysis, serum albumin ≤4.0 g/dL (HR 1.667, 95% CI 1.063–2.618, b-coefficient 0.511, P = 0.026), multi-nodularity (HR 1.750, 95% CI 1.120–2.734, b-coefficient

0.560, P = 0.014), AFP >100 ng/mL (HR 1.770, 95% CI 1.124–2.788, b-coefficient 0.571, P = 0.014) and E-cadherin downregulation (HR 4.690, 95% CI 2.700–8.144, b-coefficient 1.545, P < 0.001) were risk factors predicting HCC recurrence. A 6-point risk score system composed of these factors weighted by b-coefficients in multivariate analysis of the training set was developed and validated in another cohort. The cumulative recurrence-free survivals were clearly differentiated in patients with different risk scores. The discriminative ability predicting recurrence based on Alaike information criteria among BCLC, CLIP and the new-developed score system were 1035.712, 1028.638 and 1023.968, respectively. Conclusions: This novel scoring system had the lowest Alaike information criterion with better discriminative ability to predict postoperative HCC recurrence and may be of value in determining follow-up frequency and potential adjuvant therapy. P979 NEUTROPHIL-TO-LYMPHOCYTE RATIO IS A PREDICTOR OF ONE-YEAR SURVIVAL IN PATIENTS WITH HEPATOCELLULAR CARCINOMA RECEIVING SORAFENIB 3 A. Lue` 1 , F.J. Bustamante2 , M. Inarrairaegui ˜ , J.I. Arenas4 , M.T. Serrano1 , M. Testillano2 , S. Lorente1 , C. Gil2 , M. De la Torre3 , 4 , B. Sangro3 . 1 Servicio de Aparato Digestivo, Hospital Clinico A. Gomez ´ Lozano Blesa, Zaragoza, 2 Servicio de Aparato Digestivo, Hospital Universitario de Cruces, Bilbao, 3 Servicio de Aparato Digestivo, Clinica Universidad de Navarra, Pamplona, 4 Servicio de Aparato Digestivo, Hospital de Donostia, San Sebasti´ an, Spain E-mail: [email protected] Background and Aims: Neutrophil-to-lymphocyte ratio (NLR) is considered a prognostic factor in patients with hepatocellular carcinoma (HCC) treated by surgical resection, liver transplantation and TACE. Our aim is to investigate the prognostic significance of NLR in HCC patients treated with sorafenib. Methods: 186 consecutive patients with HCC treated with sorafenib from 4 different hospitals were enrolled between August 2005 and October 2013. Clinical, analytical and tumoral features were measured and survival was calculated from the start of sorafenib treatment. Univariate analysis and Cox regression model were used to analyze its ability to predict survival. Results: Median follow-up time was 7.2 months. 82.8% were men. Mean age was 63(±10) years. Median overall survival was 8.48 months (95% CI: 6.4–10.5). 64.6% of patients were Child A, 33.1% Child B and 2.3% Child C. 54% of patients had vascular invasion and/or metastatic disease. 46.2% had been previously treated with TACE. In univariate analysis the absence of vascular invasion (p < 0.01), BCLC stage (p < 0.05), Child class (p < 0.05), previous TACE (p < 0.01) and NRL < 2.3 (p < 0.01) were predictors of better 1-year survival. In multivariate analysis NLR < 2.3 (HR 0.55; 95% CI: 0.34–0.89; p < 0.05) and previous TACE (HR 0.56; 95% CI: 0.34–0.94; p < 0.05) were independent prognostic factors of 1-year survival. Conclusions: NLR < 2.3 and previous TACE are independent prognostic indicators for 1-year survival in patients with HCC treated with sorafenib. P980 AN EXTERNAL VALIDATION OF THE HEPATOMA ARTERIAL-EMBOLISATION PROGNOSIS (HAP) SCORE: THE LIVERPOOL EXPERIENCE O. Noorullah1 , V.P. Lekharaju2 , I.U. Din2 , J. Klcova2 , T. Cross3 , J. Evans3 , R. Sturgess2 , D. Palmer4 , P. Kumar2 , E. O’Grady2 , N. Stern2 . 1 Macclesfield Hospital, Macclesfield, 2 Aintree University Hospital, 3 Royal Liverpool Hospital NHS Trust, 4 University of Liverpool, Liverpool, United Kingdom E-mail: [email protected] Background and Aims: Most hepatocellular carcinomas (HCCs) have palliative treatment. Trans-arterial embolisation (TAE) or


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POSTERS chemoembolisation (TACE) are used with variable results. The HAP score was recently described to determine patients likely to benefit from TAE or TACE. We report our experience with TAE and TACE to assess whether the HAP score was valid for our cohort of patients. Methods: Retrospective review of cases given TAE or TACE in Liverpool, UK (2006–2013). HAP score [1 point each for albumin 400 ng/ml, bilirubin >17 mmol/l, tumour diameter >7 cm. HAP A = 0 points, B = 1, C = 2, D > 2]. Outcome recorded according to HAP score. Results: 137 patients identified having received TAE/TACE with full data to complete HAP score. Mean age 69 years, 116 (84.7%) male. 78.8% AUH, 21.2% RLUH. HAP score A: 44 (32.1%); B: 40 (29.2%); C: 32 (23.4%); D: 21 (15.3%). Overall median survival 492 days (16 months). Median survival by HAP score, A: 492 days; B: 839 days; C 478 days; D 309 days. Log rank p < 0.001. Survival at 1 year: A 62.8%; B 75%; C 59.4%, D 28.6%. Survival at 2 years: A 29.5%, B 52.5%, C 37.5%, D 14.3%. HAP D patients had lower median survival (309 vs. 563 days; p < 0.001) and 1 and 2 year survival (28.6% vs. 65.5%; p = 0.001 and 14.3% vs 39.7%; p = 0.021). Conclusions: Patients with HAP score D due TACE have a relatively poor outcome in this external validation group. This should be considered when planning treatment or further trials. P981 MODERN SURGICAL LIVER RESECTIONS OFFER BETTER RESULTS THAN TRANSARTERIAL CHEMOEMBOLIZATION IN CIRRHOTIC PATIENTS WITH B-BCLC STAGE HEPATOCELLULAR CARCINOMA R. Ciria1 , A.B. Gallardo1 , J. Cabrera1 , A. Luque1 , M.D. Ayllon ´ 1, 2 1 , J.L. Montero2 , P. Lopez-Cillero ´ , M. Rodr´ıguez-Peralvarez ´ M. de la Mata2 , J. Briceno1 . 1 Unit of Hepatobiliary Surgery and Liver Transplantation. IMIBIC, 2 Unit of Hepatology and Liver Transplantation. IMIBIC, Reina Sof´ıa University Hospital, Cordoba, Spain E-mail: [email protected] Background and Aims: BCLC-based management of B-stage HCC lies on TACE, offering a reduced survival. Our aim was to analyze the impact of liver resection (LR) versus TACE. Methods: Retrospective analysis of patients with B-BCLC HCC treated with both LR or TACE between 2006–2012 in our tertiary referral hospital. Data exposure: Mean(SD) and %. Overall survival, free-of-disease and recurrence (log-rank-test and multivariateCox). Results: Eighty patients were treated (45-TACE and 35-LR). Age was 66(10) years. Number of nodules was [1.8(1.1)]; multinodularHCC was observed in 50% of cases. Child-B (23.5%) was higher in TACE vs LR. Overall-survival was 25.81 months (median followup=20.5 months), with an overall-mortality of 55.5% vs 31.4% in TACE vs. LR, respectively. TACE vs LR 1-, 2-, and 3-years survival was 71.1%, 55.6% and 44.4% vs. 80%, 71.4% and 68.6%, respectively (P = 0.011). Overall-HCC recurrence in TACE vs. LR groups was 44.4% vs. 34%, respectively. Time-to-recurrence was 16.14(14.28) months, with a free-of-disease-time of 12.02(12.61) vs. 21.4(14.7) months in the TACE vs LR groups, respectively. Multivariate analysis (36-months survival endpoint) showed that tumour size (HR = 1.14[1.03–1.27]) and TACE (HR = 4.05[1.7–9.7]) were independent risk factors. Univariate LR analysis showed high 36-months survival (P = 0.04) in the groups with good/moderate differentiation without vascular invasion (VI) (92.3%) against the other groups (54.5%). Subgroup-multivariate analysis showed that this group was a protective factor of survival (HR = 8.774[1.12– 68.98]) and low-recurrence (HR = 10.733[1.35–85.03]). Conclusions: Management of B-BCLC HCC patients should be more complex. Modern surgical resection offers excellent survival benefit with optimal security, especially in patients with good/moderate differentiation with no vascular invasion. S402

P982 MULTIDISCIPLINARY TREATMENTS IMPROVE SURVIVAL IN PATIENTS WITH ADVANCED HEPATOCELLULAR CARCINOMA: ANALYZING THE SURVIVAL RATE OF PATIENTS AFTER SORAFENIB ADMINISTRATION N. Hattori, K. Tsuchiya, N. Nakakuki, H. Takada, S. Matsuda, S. Kaneko, M. Muraoka, N. Tamaki, Y. Yasui, R. Osaki, S. Suzuki, T. Hosokawa, H. Nakanishi, J. Itakura, Y. Takahashi, M. Kurosaki, N. Izumi. Gastroenterology and Hepatology, Musashino Red Cross Hospital, Tokyo, Japan E-mail: [email protected] Background and Aims: Median survival times (MSTs) of the patients who received sorafenib therapy in 2 phase 3 trials were 10 and 6.5 months. We analyzed the clinical features of patients who survived more than 1.5 years after sorafenib administration. Methods: Between December 2009 and August 2013, 140 patients were administered sorafenib in our center. Serial measurements of liver function, plasma VEGF, and serum AFP concentrations were evaluated at baseline and repeated at 4-week intervals. MDCT was performed one month after initial sorafenib administration and repeated at 3-month intervals. Results: Of the 140 patients, 39 survived for >1.5 years after sorafenib administration and 30 for ≤1.5 years, however the remaining 73 died. MST of these patients was 12.6 months. Pretreatment factors significantly associated with >1.5-year survival were serum albumin, AFP, AFP-L3, and major portal invasion. Although the pattern of radiological tumor progression (target lesion growth or new lesion occurrence) was a significant factor, existence of only radiological progression was not significant. Multivariate analysis revealed that VEGF of 1.5-year survival after sorafenib administration. Conclusions: The PD pattern and plasma VEGF concentration were very important factors associated with >1.5-year survival after sorafenib administration. In addition, multidisciplinary treatments should be performed to improve the survival rate in these patients after sorafenib administration. P983 VALIDATION OF THE HEPATOME ARTERIAL-EMBOLIZATION PROGNOSTIC (HAP)-SCORE IN AN AUSTRIAN TACE COHORT M. Pinter1 , F. Hucke1 , I. Graziadei2 , W. Vogel2 , M. Trauner1 , W. Sieghart1 , M. Peck-Radosavljevic1 , Vienna Liver Cancer Study Group. 1 Gastroenterology & Hepatology, Medical University of Vienna, Vienna, 2 Division of Gastroenterology and Hepatology, Medical University of Innsbruck, Innsbruck, Austria E-mail: [email protected] Background and Aims: The recently published Hepatome ArterialEmbolization Prognostic (HAP)-score was developed to facilitate the selection of HCC patients for TACE and differentiates four prognostic subgroups by means of albumin, AFP, bilirubin, and tumor diameter. The aim of this study was to externally validate the HAP-Score in an Austrian cohort of HCC patients treated with TACE. Methods: Patients diagnosed with HCC at BCLC-stage A/B treated with TACE between 01/1999 and 12/2011 at the Medical Universities of Vienna and Innsbruck were included. The following variables were entered into univariate analysis: age 65 years, sex, etiology, tumor extent, tumor number, Child–Pugh stage, creatinine, BCLCstage, and HAP-score. Variables with a p-value 2.3, bilirubin >2, sodium 200 ng/mL HR 1.95 (95% CI 1.5–2.5, p < 0.001) and ascites HR 2.16 (95% CI 1.4–3.4, p = 0.001). Conclusions: First-line treatment with DEB-TACE achieved 71% 5yr survival in BCLC A, comparable to radical therapies and it can be safely applied to advanced HCCs with peripheral neoplastic portal thrombosis without extrahepatic spread. P993 EASL AND MRECIST RESPONSE TO COMBINATION THERAPY OF SORAFENIB AND TRANSARTERIAL CHEMOEMBOLIZATION PREDICTS SURVIVAL IN HEPATOCELLULAR CARCINOMA W. Wang, W. Bai, Y. Zhao, Z. Yin, C. He, J. Niu, L. Liu, H. Chen, D. Fan, G. Han. Dept. of Liver Disease and Digestive Interventional Radiology, Xijing Hospital of Digestive Diseases, Fourth Military Medical University, Xi’an, China E-mail: [email protected] Background and Aims: There was no study investigating the use of RECIST, EASL and mRECIST for assessing treatment response to combination therapy of sorafenib and transarterial chemoembolization in patients with hepatocellular carcinoma (HCC). The aim of this study was to explore the earliest time point for accurate assessment of response to combination therapy in HCC patients, as well as to compare the three criteria and to validate the prognostic value of them when applied at this early time point post-therapy.


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POSTERS Methods: 114 consecutive patients, receiving combination therapy with sufficient imaging follow-up entered this study. RECIST, EASL and mRECIST were applied for evaluation of response and results were compared at different evaluation time points. Results: At 1–2- and 2–3-months post-therapy, no difference in OS between responders and nonresponders were observed regardless of which criterion was used. At 3–4-months, response rate obtained using EASL (50.6%) or mRECIST (51.6%) was much higher than that assessed with RECIST (16.5%). The intermethod agreement between mRECIST and EASL was perfect (k = 0.9), while it was slight (k = 0.3) between mRECIST and RECIST. EASL and mRECIST responses were significantly associated with survival. A 52% and a 50% risk reduction were observed for EASL and mRECIST responders respectively, compared to nonresponders. However, there was no significant association between treatment response and survival based on RECIST. Conclusions: 3–4-months post-treatment is considered to be the earliest time point for evaluation of response to combination therapy. EASL and mRECIST responses are independent predictors for OS when assessed at this early time point. P994 ANTIVIRAL THERAPY IMPROVES OUTCOMES IN PATIENTS WITH SMALL HEPATOCELLULAR CARCINOMA AND CHRONIC HEPATITIS C VIRUS INFECTION TREATED WITH RFA N. Zhang1 , F. Li1 , J. Liu1 , N.M. Kemmer2 , G. Neff2 , W. Lu1 . 1 Tianjin Second People’s Hospital and Tianjin Institute of Hepatology, Tianjin, China; 2 Tampa General Medical Group, Tampa, FL, United States E-mail: [email protected] Background and Aims: Chronic hepatitis C virus (HCV) associated hepatocellular carcinoma (HCC) shows a high rate of recurrence even after curative treatment. The aim of this study is to investigate the efficacy of antiviral therapy after curative radiofrequency ablation (RFA) for HCC patients infected with HCV (genotype 1). Methods: From December 2007 through December 2010, we identified 84 patients with chronic Hepatitis C (HCV) genotype 1 and HCC (tumor diameter 3 cm or less and number of tumors three or fewer). We divided the patients into two groups: control group (n = 42) received RFA only and treatment group (n = 42) received RFA and peg-IFN and weight based ribavirin. The following data was extracted; sustained virological rate (SVR), safety, tumor recurrent rate, 1-year and 3-year survival rate. Results: The 1-year survival rate between control group and treatment group was 95.2% and 90.5%, P > 0.05. The 3-year survival rate was higher in the treatment group compared with the control group (83.3% vs 66.7%, P = 0.004). The rate of initial recurrence did not differ significantly between control group and treatment group (36%, 57.1%, vs 21.4%, 47.6%, at 1, 3, years, P = 0.181,respectively), treatment group showed a lower rate than the control group for second recurrence (26.2%, 59.5%, vs 47.6%, 71.4%, at 1, 3, years, P = 0.0243, respectively). Among the treatment group, patients with sustained virologic response (SVR) were less likely to have a second HCC recurrence than the treatment group patients without an SVR, P = 0.003. Conclusions: These results indicate that successful antiviral therapy after RFA for HCV-related HCC may lower tumor recurrence rate and prolong survival.

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P995 SORAFENIB VERSUS 90 Y-RADIOEMBOLIZATION FOR ADVANCED MONO-LOBAR HEPATOCELLULAR CARCINOMA IN CIRRHOTIC PATIENTS: A PRELIMINARY STUDY ON TOLERABILITY AND SURVIVAL S. Gaia1 , A. Cantamessa1 , M. Tabone2 , D. Campion1 , P. Carucci1 , M. Grosso3 , P.R. Brunocilla1 , A. Risso1 , A. Calvo4 , E. Rolle1 , M. Vesan1 , F. Brunello1 , M. Rizzetto1 . 1 Gastro-Hepatology, A.O. Citt` a della Salute e della Scienza di Torino, University of Turin, 2 Gastroenterology, Mauriziano Hospital of Turin, Turin, 3 Radiology, S.Croce e Carle Hospital of Cuneo, Cuneo, 4 Interventional Radiology, Mauriziano Hospital of Turin, Turin, Italy E-mail: [email protected] Background and Aims: We evaluated survival and tolerability of Sorafenib and Y90 -Radioembolization (TARE) treatments in cirrhotic patients with advanced mono-lobar hepatocellular carcinoma (HCC). Methods: Data from our prospective database were analyzed. Inclusion criteria: Child–Pugh A liver cirrhosis with mono-lobar HCC; BCLC B/C; M0; N0/N1; treatment with Sorafenib >60 days or with TARE (1–2 sessions); follow-up >180 days. Liver-related complications, side effects and survival (Kaplan-Mayer curves) were analyzed. Results: Thirty-three patients treated with Sorafenib between January 2008 and November 2013 were included: 29 male; median age: 69 years (32–82); ECOG 0; median tolerated drug dose: 800 mg (200–800); median treatment time: 4.9 months (range: 2–23.4). Thirty patients undergoing TARE between May 2011 and November 2013 were included: 29 male; median age: 66.5 years (50–79); ECOG 0; treated with 1 (29/30) or 2 sessions. Side effects occurred in 94% (31/33) of Sorafenib group (mostly asthenia, hand-foot skin syndrome, diarrhea, nausea) and in 27% (8/30) of TARE group (nausea, vomiting, allergic reaction) (p < 0.001). Liver function worsened (Child–Pugh ≥B) in 24% (8/33) of Sorafenib and in 10% (3/30) of TARE group after 30–60 days from therapy (p > 0.05). Overall 44 deaths occurred (27 sorafenib, 17 TARE). Median overall survival was 11 months (95% CI: 9.6–12.4) with Sorafenib and 12 months (95% CI: 9.0–15.1) with TARE (p > 0.05). Cumulative probability of survival at 6–12–18 months was 84.3–40.6–19.0% with Sorafenib versus 86.3–48.2–33.0% with TARE (p > 0.05). Conclusions: TARE has a lower rate of side effects and similar overall survival compared to Sorafenib in cirrhotic patients with advanced mono-lobar HCC. P996 PROPOSAL FOR AN UPDATE OF THE BARCELONA STAGING SYSTEM FOR HEPATOCELLULAR CARCINOMA BASED ON THE ROLE OF PERFORMANCE STATUS A. Pecorelli1 , A. Vitale2 , L. Venerandi1 , A. Cucchetti3 , F. Trevisani1 , L. Bolondi1 , F. Piscaglia1 , The ITA.LI.CA. (Italian Liver Cancer) Study Group. 1 Department of Medical and Surgical Science, Bologna University, Bologna, 2 Padua University Hospital, Padua, 3 Department of Surgery and Transplantation, Bologna University, Bologna, Italy E-mail: [email protected] Background and Aims: Barcelona Clinic Liver Cancer (BCLC) staging system adopted the ECOG Performance Status (PS) to define stages. However, in case of PS1 it is often difficult to ascertain the tumor or cirrhosis origin, hence we aimed to assess the prognostic role of PS levels in the BCLC system. Methods: We retrospectively analyzed 2367 patients included in the ITA.LI.CA. (Italian Liver Cancer) database from 2000 (Barcelona conference) to end 2012. At first we assessed the prognostic role of different liver function and tumor related variables, including PS levels, and then we tested the prognostic capacity of the


POSTERS BCLC staging when modified according to the findings of PS or disregarding it. Results: At multivariate analysis only PS≥2 vs PS0–1, beside various liver function and tumor related variables, but not PS≥1 vs PS0 resulted significant predictors of survival. We then compared the prognostic capacity of the original BCLC staging (model 1) vs a complete removal of PS (model 2) vs a model where only PS≥2 qualifies a patient to be advanced, but not PS1, according to our findings. The discrimination ability among the various BCLC stages (AvsBvsCvsD), expressed by the Akaike information criterion (AIC), was significantly better in model 2 (943) and 3 (880) than in model 1 (699, p < 0.05). Model 3 produced the best progressive discriminatory capacity among stages. Conclusions: Patients with HCC on cirrhosis and PS1 should not be considered advanced, due to the difficulty in ascribing mild symptoms, commonly observed in cirrhosis, to the tumor. P997 NON SUPERIORITY OF DRUG-ELUTING BEADS WITH RESPECT TO CONVENTIONAL TRANSARTERIAL CHEMOEMBOLIZATION FOR THE TREATMENT OF UNRESECTABLE HEPATOCELLULAR CARCINOMA: A PROPENSITY SCORE ANALYSIS A. Facciorusso1 , C. Sposito1 , L. Mariani2 , C. Spreafico3 , A. Marchiano` 3 , S. Bhoori1 , M. Barone4 , V. Mazzaferro1 . 1 Surgery and Hepatology, 2 Medical Statistics, 3 Interventional Radiology, National Cancer Institute, IRCCS, Milan, 4 Gastroenterology, University of Foggia, Foggia, Italy E-mail: [email protected] Background and Aims: Aim of this study is to evaluate the efficacy in terms response rate, time to progression (TTP) and overall survival (OS) of drug-eluting beads (DEB) TACE in comparison with conventional (c-TACE) for the treatment of hepatocellular carcinoma (HCC). Methods: The study included 249 early/intermediate HCC patients consecutively treated with TACE between Jan 2007 and Dec 2011. Among them 145 underwent DEB-TACE (group 1) and 104 c-TACE (group 2). Treatment response was evaluated 1 month after the procedure by means of mRECIST and EASL criteria, and repeat treatment was scheduled “on demand” in case of residual viable tumor. A propensity score (PS) was generated integrating liver function and tumor related prognostic factors. Unadjusted and adjusted (according to PS) survival analysis was computed. Results: CPT status was A in 89% of cases and 58% of patients were BCLC B in both groups. The study groups were well balanced and significantly differ only according to MELD score (median MELD 8 vs 9 in group 1 vs group 2, p = 0.046). Objective response rate were 74.81% vs 89.66% (p = 0.002) according to EASL and 74.81% vs 85.34% (p = 0.039) according to mRECIST. Median TTP was 10.51 vs 14.46 months (p < 0.001) after DEB-TACE and c-TACE respectively. Median OS was 31.31 and 38.28 months in group 1 and 2 respectively (HR = 1.33; 95% CI: 0.94–1.87; p = 0.1077) and the PS-adjusted hazard ratio was 1.30 (95% CI:0.87–1.92; p = 0.1973). No significant differences in grade 3/4 adverse events were found. Conclusions: DEB-TACE is not superior to c-TACE in terms of efficacy and safety profile.

P998 BENEFIT OF SORAFENIB ACCORDING TO UNDERLYING LIVER DISEASE ETIOLOGY IN PATIENTS WITH ADVANCED HEPATOCELLULAR CARCINOMA (HCC) M. Bouattour1 , N. Goutte1 , M.-P. Vullierme2 , C. Castelnau1 , O. Farges3 , S. Faivre4 , F. Durand1 , L. Castera1 . 1 Hepatology, 2 Radiology, 3 Surgery, 4 Oncology, Beaujon Hospital, Assistance Publique-Hˆ opitaux de Paris, Clichy, France E-mail: [email protected] Background and Aims: Sorafenib is currently the only approved targeted therapy for advanced HCC. The influence of the underlying liver disease etiology on sorafenib benefit remains, however, debated. Aim: To compare benefit of sorafenib, including time to progression (TTP) and overall survival (OS), according to the underlying liver disease etiology. Methods: All patients treated by sorafenib for advanced HCC between Jan 2006 and Oct 2010 were retrospectively selected from our prospective database. Epidemiological data, liver disease etiology, prior HCC therapy, objective response (RECIST), OS and TTP were evaluated and analyzed according to underlying liver disease etiology (Alcohol, NASH, HBV and HCV). Results: 134 patients (median age 63.4 years; male 80%; BCLC stage B 25% C 75%; HBV 17%, HCV 25%, alcohol 32%, NASH 13% and miscellaneous 13%) were studied. Demographic characteristics, Child–Pugh class, BCLC and prior therapy and median duration of sorafenib (6.4 months) were similar for each subgroup. Partial response was observed in 2 patients only. The median OS was 9.5 months (95% CI: 7.35–11.67) and did not differ according to etiology (alcohol 14.2; NASH 9.9; HCV 8.4 and HBV 6.1 months; p= NS). Conversely, median TTP was significantly shorter in HBV patients compared to HCV patients (4.4 vs. 7.3 months, respectively; p = 0.036), but not between alcoholic patients and those with NASH (6.4 vs. 5.2 months; respectively; p = 0.7). Conclusions: Tumor progression under sorafenib seems to occur earlier in HBV patients compared to other etiologies. P999 Y90-RADIOEMBOLIZATION FOR INTERMEDIATE/ADVANCED HCC PATIENTS OUTSIDE THE CONVENTIONAL CRITERIA MAY BE DETRIMENTAL: A SINGLE CENTER EXPERIENCE C. Sposito1 , A. Facciorusso1 , T. Camerini2 , C. Chiesa3 , M. Maccauro3 , C. Morosi4 , S. Bhoori1 , D. Citterio1 , V. Mazzaferro1 . 1 Gastrointestinal Surgery and Liver Transplant Unit, Fondazione IRCCS Istituto Nazionale Tumori, Milano, 2 Medical Statistics, 3 Nuclear Medicine, Fondazione IRCCS Istituto Nazionale Tumori, Milan, 4 Interventional Radiology, Fondazione IRCCS Istituto Nazionale Tumori, Milano, Italy E-mail: [email protected] Background and Aims: Transarterial radioembolization (TARE) represents a promising treatment for intermediate/advanced HCC. A group of experts has recently identified as contraindications to TARE decompensated cirrhosis (CPT ≥B8), tumor burden >50% and ECOG PS ≥1. Aim of this study is to evaluate the impact of this variables on patients’ survival. Methods: Data of 118 HCC patients consecutively treated with TARE (Therasphere® ) between Dec 2010 and Dec 2012 were reviewed. Univariate and multivariate Cox Regression analysis was performed for overall survival (OS) and progression-free survival (PFS), evaluating the impact of liver function and tumor related parameters. Liver decompensation rate within 6 months was analyzed. Results: Most of patients were CPT A (88.1%), 66.1% were BCLC C with portal vein thrombosis (PVT) and 89.8% had ≤50% of tumor burden. Performance status was 0 in 98% of cases.


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POSTERS Median PFS and OS were 7 and 11 months respectively. Liver decompensation within 6 months occurred in 36.6% of patients. Overall survival in CPT A patients was 11 months vs 6.5 in CPT B (p = 0.0016), while in patients with liver involvement ≥50% was 7 months vs 15 in case of more limited tumor burden (p = 0.026). Patients with PVT reached an OS of 10 months vs 17 months in BCLC B HCCs (p = 0.047). At multivariate analysis, the only factors significantly related to OS were CPT stage, tumor burden and presence of PVT. Conclusions: TARE is an effective safe procedure, but should be contraindicated in presence of deteriorated liver function and massive tumor burden. P1000 CLINICAL OUTCOMES AND SAFETY PROFILE OF TRANSARTERIAL CHEMOEMBOLIZATION FOR HEPATOCELLULAR CARCINOMA WITH SMALL (70–150 mM, M1) DRUG ELUTING BEADS: A PHASE I STUDY C. Spreafico1 , C. Sposito2 , T. Cascella1 , A. Facciorusso2 , M. Bongini2 , S. Bhoori2 , V. Mazzaferro2 . 1 Interventional Radiology, 2 Gastrointestinal Surgery and Liver Transplant Unit, Fondazione IRCCS Istituto Nazionale Tumori, Milano, Italy E-mail: [email protected] Background and Aims: The aim of this prospective phase I study is to assess the efficacy and the safety profile of TACE using 70– 150 mm drug eluting beads (DC BeadM1-TACE, Biocompatibles UK), a new embolizing device that may allow for more concentrated drug delivery and greater tumoral devascularisation. Methods: The study included 45 consecutive HCC patients recruited between July 2012 and October 2013 who underwent DC BeadM1TACE. Clinical data were recorded at 12, 24 and 48 h, 7 and 30 days after the treatment. Response was assessed by CT-scan after one month according to mRECIST and EASL criteria and a second BC Bead M1-TACE within six weeks was scheduled in case of noncomplete response. Results: The median age was 61 years, the main etiology was HCV (44.4%) and CPT status was A in 97.7% of cases. Tumor stage was BCLC B in 44.4% and BCLC A in the remaining cases; the median number of nodules and their sum of diameters were 2 (range 1–6) and 4.3 cm (10–190) respectively. Overall, 63 treatments were performed. A complete response was observed in 31.1% of patients and 28.9% showed a partial response with according both to mRECIST and EASL criteria (Figure 1), with a median time to response of 3 months (IC 95% 2–4). The treatment was well-tolerated and only 2 (4.4%) patients experienced severe (grade 3/4) adverse events according to CTCAE 4.0.

Figure 1. Waterfall diagram of best response after M1 TACE according to mRECIST.

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Conclusions: Chemoembolization using DC BeadM1 is an effective procedure with a favorable safety profile and interesting results in terms of objective response rate. P1001 CLINICAL OUTCOMES AFTER RESECTION IN PATIENTS WITH NASH-RELATED HCC R. Dhanasekaran1 , J.D. Yang1 , N. Giama1 , S.W. Slettedahl2 , W.S. Harmsen2 , T. Therneau2 , L.R. Roberts1 . 1 Gastroenterology and Hepatology, 2 Mayo Clinic Rochester, Rochester, MN, United States E-mail: [email protected] Background and Aims: Nonalcoholic fatty liver disease (NAFLD) is the most common causes of chronic liver disease in Western countries and it has also been partially implicated in the rising incidence of hepatocellular carcinoma (HCC). Our aim was to analyze a series of patients with NASH related early stage HCC who underwent resection. Methods: Clinical, laboratory and survival data were collected for atients with NASH who were diagnosed with HCC at Mayo Clinic, Rochester between 2005 and 2011. Results: Total of 117 patients had NASH-HCC; 13.7% underwent resection. Components of metabolic syndrome were common, hypertension (87.5%), diabetes (56.2%) and hyperlipidemia (44%). Majority (75%) had BMI >25 and 25% had medically-complicated obesity. All patients had Child–Pugh A disease without portal hypertension. Histologically, most patients (81.5%) had fibrosis stage 38.0 35.0–38.0 180 100–180 F2) and 34 (12.6%) having cirrhosis (F4). The AUC for significant fibrosis and cirrhosis for ANN model were 0.89 (95% CI 0.85–0.93) and 0.94 (%95 CI 0.91–0.98), respectively. ANN probability scores (range 0–1.0) 0.90 had negative and positive predictive values of 97% and 92%. Restricting the liver biopsy in patients with intermediate scores may prevent liver biopsy in 39% of patients with >95% accuracy. The areas under the ROC curve for significant fibrosis for AAR, APRI, GUCI, FORNS, HALT-C, HUI and FIB-4 scores were 0.67, 0.87, 0.84, 0.80, 0.80, 0.78 and 0.84 respectively. Correspondingly, AUCs for cirrhosis or AAR, APRI, GUCI, FORNS, HALT-C, HUI and FIB-4 scores were 0.75, 0.84, 0.86, 0.80, 0.85, 0.79 and 0.87. Conclusions: ANN model based on routinely available biochemical and ultrasound parameters is an accurate and cheap option for prediction of significant liver fibrosis and cirrhosis in chronic hepatitis B with favorable diagnostic accuracy in comparison with other free noninvasive scores. P1007 SOLUBLE CD163, A MACROPHAGE ACTIVATION MARKER, IS INDEPENDENTLY ASSOCIATED WITH FIBROSIS IN PATIENTS WITH CHRONIC VIRAL HEPATITIS B AND C K. Kazankov1 , F. Barrera2 , H.J. Møller3 , B.M. Bibby4 , H. Vilstrup1 , J. George2 , H. Grønbæk1 . 1 Department of Hepatology and Gastroenterology, Aarhus University Hospital, Aarhus, Denmark; 2 The Storr Liver Unit, Westmead Millenium Institute, University of Sydney and Westmead Hospital, Westmead, NSW, Australia; 3 Department of Clinical Biochemistry, Aarhus University Hospital, 4 Department of Biostatistics, Aarhus University, Aarhus, Denmark E-mail: [email protected] Background and Aims: Macrophages are involved in inflammation and liver fibrosis. Soluble (s)CD163 is a specific marker of activated macrophages. Methods: We investigated associations between sCD163 and biochemical and histological parameters of inflammatory activity and fibrosis in 556 patients with chronic hepatitis C virus (HCV) and 208 patients with chronic hepatitis B virus (HBV) before anti-viral treatment. Scheuer histological scores of activity and fibrosis were obtained. Clinical, biochemical, and metabolic parameters were recorded. We measured sCD163 by enzyme-linked immunosorbent assay (ELISA). Results: Soluble CD163 was higher in patients with HCV compared to HBV [3.7 (interquartile range (IQR) 2.5–5.5) vs. 2.4 (IQR S410

1.8–3.6) mg/L, p < 0.001). sCD163 was associated with fibrosis stages, with highest levels in patients with advanced fibrosis (F ≥ 3) [5.7 (IQR 3.9–6.9) mg/L] and cirrhosis (F = 4) [6.2 (IQR 5.0–6.9) mg/L]. sCD163 was a marker of fibrosis independent of other biochemical parameters and known risk factors. A sCD163-based Fibrosis Score, CD163-FS, had an area under the Receiver Operating Characteristics curve (AUROC) of 0.87 (95% CI: 0.82–0.92) for cirrhosis, 0.84 (95% CI: 0.80–0.88) for advanced fibrosis, and 0.78 (95% CI: 0.74–0.81) for significant fibrosis. Compared to existing fibrosis scores, CD163-FS was superior to the AST to platelet ratio index (APRI) for all fibrosis stages and to FIB-4 for significant fibrosis. Conclusions: sCD163 levels are increased in patients with chronic viral hepatitis, reflecting macrophage activation. Increased sCD163 is associated with the severity of disease and predicts fibrosis. A sCD163-based fibrosis score, CD163-FS, is superior to APRI and FIB-4 for the diagnosis of significant fibrosis. P1008 A 5.9 kDa SERUM FRAGMENT OF FIBRINOGEN a-CHAIN IS A SPECIFIC SURROGATE MARKER OF ACTIVE FIBROGENESIS IN PATIENTS WITH LIVER DISEASE S. Marfa1 , G. Crespo2 , V. Reichenbach1 , X. Forns2 , G. Casals1 , 1,3 1 ´ . Biochemistry and M. Morales-Ruiz1 , M. Navasa2 , W. Jimenez Molecular Genetics Service, 2 Liver Unit, Hospital Cl´ınic, IDIBAPS, CIBEREHD, 3 Departament de Ciencies Fisiologiques I, University of Barcelona, Barcelona, Spain E-mail: [email protected] Background and Aims: Early detection of fibrosis progression is of major relevance for the diagnosis and management of patients with liver disease. This study was designed to find noninvasive biomarkers of fibrosis in a clinical context in which this process occurs rapidly, HCV-positive patients undergoing liver transplantation (LT). Methods: We analyzed 93 LT patients with HCV recurrence, 17 non-LT patients with liver disease and 9 LT patients without HCV recurrence who received antiviral treatment before LT, as the control group. Blood obtained from 5 healthy subjects was also analyzed. Serum samples were fractionated by ion exchange chromatography and their proteomic profile was assessed by SELDITOF-MS. Results: Marked differences were observed between the serum proteome profile of LT patients with early fibrosis recurrence and non-recurrent LT patients. A robust peak intensity located at 5905 m/z was the distinguishing feature of non-recurrent LT patients. However, the same peak was barely detected in recurrent LT patients. Similar results were found when comparing samples of healthy subjects with those of non-LT fibrotic patients, indicating that our findings were not related to either LT or HCV infection. Using tandem mass-spectrometry, we identified the protein peak as a C-terminal fragment of the fibrinogen a chain. Cell culture experiments demonstrated that TGF-b reduces a-fibrinogen mRNA expression and 5905 m/z peak intensity in HepG2 cells, suggesting that TGF-b activity regulates the circulating levels of this protein fragment. Conclusions: In conclusion, we identified a 5.9 kDa C-terminal fragment of the fibrinogen a chain as an early serum biomarker of fibrogenesis in patients with liver disease.


POSTERS P1009 THE HISTOLOGICAL OR ULTRASONOGRAPHIC DETECTION OF STEATOSIS AFFECTS THE PERFORMANCE OF LIVER STIFFNESS MEASUREMENT FOR FIBROSIS ASSESSMENT IN PATIENTS WITH CHRONIC HCV GENOTYPE 1 INFECTION F.S. Macaluso1 , M. Maida1 , C. Camma` 1 , G. Cabibbo1 , D. Cabibi2 , V. Di Marco1 , A. Craxì1 , S. Petta1 . 1 Sezione di Gastroenterologia, Di.Bi.M.I.S., 2 Cattedra di Anatomia Patologica, University of Palermo, Palermo, Italy E-mail: [email protected] Background and Aims: In Chronic Hepatitis C (CHC), the influence of steatosis on liver stiffness measurement (LSM) is still debated. We assessed the impact of steatosis and its ultrasonographical sign – i.e. Bright Liver Echo Pattern (BLEP) – on LSM values and on Transient Elastography (TE) accuracy for the diagnosis of liver fibrosis, in a cohort of consecutive biopsy-proven patients with Genotype 1 (G1) CHC. Methods: Consecutive G1 CHC patients (n = 618), assessed by demographic, biochemical, metabolic, ultrasonographic and histological features, were included. TE was assessed using the standard M probe. Results: Male gender (p = 0.04), steatosis as continuous variable (p < 0.001), severity of necroinflammation (p = 0.02) and stage of fibrosis (p < 0.001) were associated with LSM by multivariate linear regression analysis. Among patients within the same fibrosis stages (F0–F2 and F3–F4; F0–F3 and F4), mean LSM values, expressed in kPa, were significantly higher in subjects with moderate-severe steatosis (≥ 20%) compared with those without, as well as in patients with BLEP on ultrasonography (US) compared with those without. Furthermore, in subjects without severe fibrosis (F0–F2) and without cirrhosis (F0–F3), a higher rate of false-positive LSM results was observed in patients with steatosis ≥ 20% (F0–F2: 35.3% vs. 17.9%; F0–F3: 38.9% vs. 16.6%) and in patients with BLEP on US (F0–F2: 28.0% vs. 18.3%; F0–F3: 29.7% vs. 17.8%) compared with their counterparts. Conclusions: In patients with G1 CHC, the presence of moderatesevere steatosis, detected by histology or by US, should always be taken into account in order to avoid overestimations of liver fibrosis assessed by LSM. P1010 LIVER STIFFNESS IN HCV AND ALD: FIBROSIS-RELATED CUT-OFF VALUES DEPEND ON DEGREE AND LOCATION OF INFLAMMATION S. Mueller1 , S. Englert2 , B. Boozari3 , H.K. Seitz1 , M. Beaugrand4 , M. Platon Lupsor5 . 1 Department of Internal Medicine, Salem Medical Center and Center for Alcohol Research, University of Heidelberg, 2 Institute of Medical Biometry and Informatics, University of Heidelberg, Heidelberg, 3 Department of Medicine I, University of T¨ ubingen, T¨ ubingen, Germany; 4 Hepatology Department, Hospital Jean Verdier, APHP, University of Paris 13, Paris, France; 5 Department of Ultrasonography, University of Medicine and Pharmacy ‘Cluj-Napoca’, Cluj-Napoca, Romania E-mail: [email protected] Background and Aims: Liver stiffness (LS) can be drastically elevated during liver inflammation making it difficult to separate fibrosis- from inflammation-induced LS. We here performed a multicenter study both on a portal-tract pronounced (hepatitis C virus infection; HCV) and a predominantly lobular disease (alcoholic liver disease; ALD). The major aim was, first, to learn whether portal and lobular inflammation differently translate into LS elevation and, second, to calculate adapted cut-off values for concomitant inflammation. Methods: 452 patients with ALD and 1393 patients with HCV were enrolled from five centers (Heidelberg, Paris, Hannover, Dusseldorf, ¨ Cluj Napoca). All patients had histological-proven fibrosis stage

F0–F4 (METAVIR or Kleiner), LS (Fibroscan) measurement and lab tests. Results: Among the routine laboratory parameters for liver damage, GOT correlated best with LS (r = 0.54, P < 10E-90) both for HCV and ALD. Therefore, we next calculated the area under the receiver operating curves (AUROC) as a function of GOT levels. Interestingly, cut-off values determined at optimal sensitivity and specificity (Youden index) were almost identical for F0, F3 and F4 in the absence of elevated transaminases (HCV: 5.1, 9.0 and 11.9 kPa, ALD: 4.9, 8.1 and 10.5 kPa). These cut-off values increased exponentially as a function of GOT levels. The impact of GOT on LS was higher in lobular-pronounced inflammation (ALD) and at higher fibrosis stages. Consideration of GOT improved AUROCs for both diseases. Conclusions: We propose novel GOT-adapted cut-off values namely for HCV were short interventions such as alcohol detoxication are not applicable. P1011 ACOUSTIC RADIATION FORCE IMPULSE-IMAGING IN COMPARISON TO TRANSIENT ELASTOGRAPHY FOR NON-INVASIVE ASSESSMENT OF LIVER FIBROSIS IN CHRONIC HEPATITIS C: A PROSPECTIVE INTERNATIONAL MULTICENTER STUDY M. Friedrich-Rust1 , M. Lupsor-Platon2 , R. de Knegt3 , V. Dries4 , P. Buggisch5 , M. Gebel6 , B. Maier1 , E. Herrmann7 , A. Sagir8 , R. Zachoval9 , Y. Shi1 , M.D. Schneider1 , R. Badea2 , K. Rifai6 , S. Zeuzem1 , C. Sarrazin1 . 1 Department of Internal Medicine 1, J.W. Goethe University Hospital, Frankfurt, Germany; 2 Department of Medical Imaging, Regional Institute of Gastroenterology and Hepatology ‘Octavian Fodor’, Cluj-Napoca, Romania; 3 Department of Gastroenterology and Hepatology, Erasmus MC University Medical Center, Rotterdam, Netherlands; 4 Institute of Pathology, Mannheim, 5 Institute for Interdisciplinary Medicine, Hamburg, 6 Department of Gastroenterology and Hepatology, Medizinische Hochschule Hannover, Hannover, 7 Institute of Biostatistics and Mathematical Modelling, Faculty of Medicine, J.W.Goethe University, Frankfurt, 8 Department of Gastroenterology and Hepatology, University Hospital D¨ usseldorf, Duesseldorf, 9 Department of Internal Medicine II, Ludwig-Maximilian University Hospital, Munich, Germany E-mail: [email protected] Background and Aims: Acoustic Radiation Force Impulse (ARFI)Imaging (Siemens Acuson S2000) is an ultrasound-based elastography method which is integrated in a conventional ultrasound machine enabling the exact localisation of measurement site. It might present an alternative method to transient elastography (TE, FibroScan® , Echosens) for the non-invasive assessment of liver fibrosis. The aim of the present prospective international controlled multicenter study was to directly compare ARFI and TE for the assessment of liver fibrosis in patients with chronic hepatitis C using liver biopsy as reference method. Methods: ARFI-imaging involves the mechanical excitation of tissue resulting in shear-wave propagation which is tracked using ultrasonic, correlation-based methods and is recorded in m/s. 253 patients with chronic hepatitis C were prospectively included in the present study and received ARFI-imaging, TE and blood tests. Liver histology was staged by a central pathologist and used as reference method. In addition, 24 patients with liver cirrhosis confirmed by imaging methods were included. Results: Finally, 247 patients could be included in the intention to diagnose analysis and 182 patients in the per protocol analysis. ARFI imaging and TE correlated significantly with histological fibrosis stage. The diagnostic accuracy expressed as areas under ROC curves for ARFI-imaging and TE are shown in Table 1. No significant difference was found between TE and ARFI in the per protocol analysis for the diagnosis of F ≥ 2 (p = 0.15), F≥3 (p = 0.11) and F=4 (p = 0.19).


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POSTERS Table (abstract P1011): AUROC for ARFI and TE Methods

F≥1

pa

F≥2

ARFI-imaging (per protocol, n= 182) FibroScan (per protocol, n = 182) ARFI-imaging (Intention to Diagnose, n = 247) FibroScan (Intention to Diagnose, n = 247)

0.75 (0.67; 0.83) 0.80 (0.72; 0.88) 0.69 (0.60; 0.79) 0.76 (0.70; 0.83)

0.28 0.28 0.21 0.21

0.81 0.85 0.77 0.85

a

(0.74; 0.88) (0.80; 0.91) (0.71: 0.83) (0.80; 0.89)

pa

F≥3

0.15 0.15 0.032 0.032

0.88 0.92 0.83 0.88

(0.82; 0.94) (0.89; 0.97) (0.77; 0.89) (0.83; 0.93)

pa

F=4

0.11 0.11 0.15 0.15

0.89 0.94 0.84 0.89

pa (0.83; 0.96) (0.90; 0.98) (0.77; 0.91) (0.84; 0.95)

0.19 0.19 0.18 0.18

p value for comparison ARFI–TE.

Conclusions: ARFI-imaging and TE are comparable methods for non-invasive staging of liver fibrosis in patients with chronic hepatitis C. P1012 ENHANCED LIVER FIBROSIS (ELF) TEST PERFORMS BETTER THAN HISTOLOGICAL PARAMETERS IN PREDICTING CLINICAL OUTCOMES IN PATIENTS WITH ADVANCED FIBROSIS DUE TO CHRONIC HEPATITIS C INFECTION G.E. Dolman1 , A.M. Zaitoun2 , W.L. Irving1 , I.N. Guha1 . 1 Nottingham Digestive Diseases Biomedical Research Unit, 2 Department of Histopathology, Queen’s Medical Centre, Nottingham, United Kingdom E-mail: [email protected] Background and Aims: Prognostic tests in chronic hepatitis C (CHC) are required to stratify patients at greatest risk of progression to a clinical outcome. Our aim was to determine the predictors of progression in a cohort of patients with advanced fibrosis. Methods: The study cohort was derived from the Trent Study of Patients with Hepatitis C. Inclusion criteria were: a) liver biopsy before 2011 demonstrating advanced fibrosis (Ishak stage ≥3); b) no clinical outcome prior to biopsy; and c) patient did not achieve sustained viral response during follow-up. Sera collected within 6 months of the index biopsy were analysed for the enhanced liver fibrosis (ELF) panel which consists of hyaluronic acid, PIIINP and TIMP-1. This was compared to liver biopsies staged by the Ishak score and quantified for collagen using automated image analysis. A clinical outcome was defined as the first event of: ascites, encephalopathy, variceal haemorrhage, hepatocellular carcinoma, transplant or liver-related death.

Results: 136 patients were identified and 87 had sera available for ELF. 33.3% patients progressed to a clinical outcome (median followup 7.2 years). ELF was significantly associated with progression to clinical outcomes in univariate analysis (HR 2.07 [95% CI 1.54–2.76]; p < 0.001). In a multivariate model including liver function tests, Ishak stage and collagen quantification, only ALP and ELF remained statistically significant (ALP: HR 1.004 [95% CI 1.001–1.007; p = 0.016], ELF: HR 1.968 [95% CI 1.454–2.663; p < 0.001]. Conclusions: Our data suggest the ELF panel can stratify risk of subsequent progression to clinical outcomes in advanced fibrosis secondary to CHC. P1013 MEASUREMENT OF LIVER AND SPLEEN STIFFNESS BY ARFI ELASTOGRAPHY: INTRA- AND INTER-OBSERVER VARIABILITY AND PREDICTORS OF VARIABILITY M. Balakrishnan1,2 , F. Souza1,2 , C. Munoz1,2 , S. Augustin1,2 , N. Loo1,2 , Y. Deng3 , M. Ciarleglio3 , G. Garcia-Tsao1,2 . 1 Digestive Diseases Section, Yale University School of Medicine, New Haven, 2 Digestive Diseases Section, VA-CT Healthcare System, West Haven, 3 Yale Center for Analytical Sciences, Yale University School of Public Health, New Haven, CT, United States E-mail: [email protected] Background and Aims: Measuring liver stiffness (LS) and spleen stiffness (SS) is a useful non-invasive way to assess fibrosis stage and portal hypertension in patients with chronic liver disease. One method to assess stiffness is elastography via acoustic radiation force impulse (ARFI) imaging. Its advantage is that sites where stiffness will be measured can be visualized ultrasonographically. However, its reliability has not been well established. We aimed to characterize the intra- and inter-observer variability of ARFImeasured LS and SS. Methods: Two hepatologists, blinded to clinical information and each other’s findings, evaluated unselected patients from outpatient liver clinics with ARFI (ACUSON S2000™; Siemens Medical Solutions, Mountain View, CA, USA). Exclusions were hepatocellular carcinoma, ascites, surgical shunt/TIPS, portal thrombosis, and cholestatic disease. Each operator obtained 20 measurements from the right liver lobe and spleen; and 10 measurements from the left liver lobe. Intercorrelation coefficients (ICC) were calculated. Table: Interobserver variability by different variables Variable

R Liver Lobe ICC (95% CI)

L Liver Lobe ICC (95% CI)

Spleen ICC (95% CI)

0.91 (0.83, 00.95)

0.83 (0.66, 0.91)

0.72 (0.51, 0.84)

0.73 (0.51, 0.86)

0.89 (0.76, 0.96)

0.75 (0.56, 0.86)

0.89 (0.79, 0.94) 0.73 (0.51, 0.86)

0.81 (0.56, 0.92) 0.90 (0.78, 0.95)

0.74 (0.54, 0.87) 0.79 (0.61, 0.89)

0.72 (0.53, 0.84) 0.88 (0.78, 0.94)

0.82 (0.64, 0.91) 0.77 (0.52, 0.90)

0.50 (0.21, 0.70) 0.80 (0.62, 0.89)

0.76 (0.57, 0.88) 0.86 (0.76, 0.92)

0.81 (0.58, 0.92) 0.83 (0.66, 0.92)

0.53 (0.22, 0.75) 0.78 (0.63, 0.88)

BMI 105 cm Spleen (length) ≤12 cm >12 cm Spleen (AP diameter) 0.3) and AILD aetiology were significantly more common in AHV (p = 0.01, 0.007, 1.4 m/s, ARFI elastography had 83.1% accuracy (AUROC = 0.822) to differentiate between subjects with or without chronic liver disease, while the best ElastPQ cut-off value to discriminate between these two categories of subjects was >1.23 m/s, with 83.7% accuracy (AUROC = 0.851). The AUROCs of ARFI elastography and ElastPQ for predicting the presence of liver diseases were similar (p = 0.48). Conclusions: Both pSWE techniques have very good feasibility for LS assessment and a good performance for predicting the presence of liver pathology.


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POSTERS P1017 EVALUATION OF THE PROGNOSTIC VALUE OF NON-INVASIVE LIVER FIBROSIS TESTS TO PREDICT MORTALITY BY TIME-DEPENDENT ROC CURVES S. Bertrais1 , J. Boursier1,2 , F. Oberti2 , I. Fouchard-Hubert2 , P. Cales ` 1,2 . 1 Faculty of Medicine, HIFIH Research Unit, UPRES 3859, SFR 4208, University of Angers, 2 Liver-Gastroenterology Department, University Hospital of Angers, Angers, France E-mail: [email protected] Background and Aims: Recent studies showed the prognostic value of some non-invasive liver fibrosis tests to predict survival and complications. In this study, we compared the ability of the liver fibrosis tests to predict mortality using time-dependent ROC curve analysis for censored survival data and calculating their AUC at several time points. Methods: 1,450 patients with various chronic liver diseases, Fibroscan, APRI, FIB-4, Hepascore, FibroMeter2G and MELD at baseline were included in a prospective cohort from 2005 to 2009. Date and causes of death were obtained from the national registry. Statistical analyses were performed using the R package timeROC. When studying liver-related mortality, other causes of death were considered as competing risks. Results: Baseline values of all liver fibrosis tests (except APRI) had good prognostic performance: AUC(t) >0.70 to predict all-cause death that occurred between 0 and 5 years, AUC(t) were from 0.80 to 0.90 for liver-related mortality. AUC(t = 3) for predicting 3-year overall mortality were: 0.80 (95% CI: [0.76–0.83]) for FibroMeter2G , 0.77 [0.74–0.81] for Hepascore (p < 0.001 vs FibroMeter2G ), 0.76 [0.73–0.80] for Fibroscan (p < 0.028), 0.76 [0.72–0.79] for FIB4 (p < 0.001) and 0.67 [0.62–0.71] for APRI (p < 0.001). The prognostic performance of FibroMeter2G was also significantly higher than those of MELD for a death prediction over 3 years. For liver-related mortality, FibroMeter2G had higher performance than MELD to predict liver-related death over 2 years, whereas Fibroscan and Hepascore were more performant than MELD for prediction ≥ 3 years. Conclusions: The time-dependent AUC comparisons provide more comprehensive information about the differences in prognostic ability between non-invasive liver fibrosis tests. P1018 MARKERS OF COLLAGEN REMODELING DETECT CLINICALLY SIGNIFICANT FIBROSIS IN CHRONIC HEPATITIS C PATIENTS M.J. Nielsen1,2 , K. Kazankov3 , D.J. Leeming1 , M.A. Karsdal1 , A. Krag2 , F. Barrera4 , J. George4 , H. Grønbæk3 . 1 Fibrosis Biology and Biomarkers, Nordic Bioscience, Herlev, 2 Department of Gastroenterology, Odense University Hospital, University of Southern Denmark, Odense, 3 Department of Hepatology and Gastroenterology, Aarhus University Hospital, Aarhus, Denmark; 4 Storr Liver Unit, Westmead Millenium Institute, University of Sydney and Westmead Hospital, Sydney, NSW, Australia E-mail: [email protected] Background and Aims: A Metavir Fibrosis stage F ≥ 2 is considered clinically significant due to progressive disease and also represent a threshold for treatment. Accumulation and change of the protein profile within the extracellular matrix (ECM) is a hallmark of liver fibrosis. ECM remodeling generates protein fragments, reflecting the turnover processes involved in liver fibrosis. We aimed to assess the diagnostic value of two serological ECM markers to detect clinically significant fibrosis. Methods: Pro-C3 (Type III collagen formation) and C4M (type IV collagen degradation) markers were assessed in plasma from 403 chronic hepatitis C (HCV) patients. Patients were stratified according to Metavir Fibrosis stage (F0–F4); F0 (n = 46), F1 (n = 161), F2 (n = 95), F3 (n = 44) and F4 (n = 33). S414

Results: Pro-C3 and C4M correlated to F stages (R = 0.48, p < 0.001 and R = 0.24, p < 0.001, respectively). Pro-C3 and C4M differed between individual fibrosis stages (p < 0.001). The diagnostic value of the two markers when separating F0–F1 from F2–F4 was significant (Pro-C3 AUC=0.75; C4M AUC=0.57). Combination of the markers with age, BMI and gender in a linear regression model increased the correlation to F stages (R = 0.60, p < 0.0001). Model levels were significantly higher in HCV patients with F≥1 compared to F0 (p < 0.0001). The diagnostic value of the algorithm was highly significant for separating F0–F1 from F2–F4 (AUC=0.80, p < 0.0001). Conclusions: Pro-C3 provided strong diagnostic accuracy as a single marker of liver fibrosis. In a model including the two markers along with patient’s age, BMI and gender the diagnostic value for identifying clinically significant fibrosis increased further. P1019 VALIDATION AND COMPARISON OF NON-INVASIVE MARKERS OF LIVER FIBROSIS IN WEST-AFRICAN PATIENTS WITH CHRONIC HEPATITIS B LIVING IN THE GAMBIA M. Lemoine1,2 , Y. Shimakawa2,3 , R. Goldin4 , M. Khalil5 , J. Lloyd4 , P. Suso5 , S. Nayagam1 , H. Freeya Njai2 , M. Taal6 , G. Ndow2 , U. D’Alessandro2 , R. Njie2 , M. Thursz1 . 1 Liver Centre, Imperial College London, St Mary’s Hospital, London, United Kingdom; 2 Disease Control and Elimination Theme, Medical Research Council, The Gambia Unit, Fajara, Gambia; 3 London School of Hygiene & Tropical Medicine, 4 Centre for Pathology, Imperial College London, St Mary’s Hospital, London, United Kingdom; 5 Department of Histopathology, Edward Francis Small Teaching Hospital, Banjul, 6 Ministry of Health, Fajara, Gambia E-mail: [email protected] Background and Aims: Non-invasive tests of liver fibrosis have been poorly studied in chronic hepatitis B in sub-Saharan Africa. In this study, using liver biopsy as a gold standard, we assessed the diagnostic accuracy of transient elastography (TE), APRI and FIB-4 for evaluating the degree of liver fibrosis in HBV-infected patients enrolled in the PROLIFICA programme in The Gambia. Methods: Between August 2012 and September 2013, all consecutive patients undergoing liver biopsy had a fasting TE (Fibroscan® ) on the same day. Patients with HIV/HCV/HDV coinfections, ascites, liver mass, pregnancy, heart failure, tuberculosis, acute infection and/or invalid liver stiffness measurements (LSM) were excluded from the final analysis; only good quality and concordant pathological results according to two independent blinded pathologists were analysed.

Figure: AUROC for predicting fibrosis ≥F2.

Results: 152 liver biopsies were performed: 18 patients did not fill the inclusion criteria and 26 (17%) samples were poor quality or discordant; 108 patients were analysed: male (70%), median age: 35 years (29–39), median LSM: 7.4 kPa (6.2–9.6), Metavir Fibrosis:


POSTERS F0–1:66 (61%), F2–4:42 (39%). For TE, APRI and FIB-4, the AUROC curves were 0.89, 0.80 and 0.73, respectively, for predicting ≥F2; 0.91, 0.79, 0.75 for ≥F3 and 0.98, 0.79 and 0.78 for F4. For TE diagnostic performances were higher compared with those of APRI (P = 0.006) and FIB-4 (P = 0.004) with optimal cut-off values at 7.5, 8.3 and 9 kPa for the diagnosis of F2, F3 and F4 respectively. Conclusions: In West-Africa, TE, APRI and FIB-4 are reliable for detection of fibrosis/cirrhosis in HBV-infected patients, TE having the best performances with slightly different cut-offs compared to Western studies. P1020 VOLATILE ORGANIC COMPOUNDS IN EXHALED AIR AS POTENTIAL NON-INVASIVE BIOMARKER FOR LIVER CIRRHOSIS K.E. Pijls1,2 , A. Smolinska3,4 , D.M.A.E. Jonkers1,2 , E.J.C. Moonen2,3 , J.W. Dallinga2,3 , A.A.M. Masclee1,2 , G.H. Koek1,2 , F.J. van Schooten2,3 . 1 Division Gastroenterology-Hepatology, Department of Internal Medicine, Maastricht University Medical Center, 2 School for Nutrition, Toxicology and Metabolism (NUTRIM), Maastricht University Medical Center, 3 Department of Toxicology, Maastricht University, Maastricht, 4 Top Institute Food and Nutrition, Wageningen, Netherlands E-mail: [email protected] Background and Aims: Early diagnosis of liver cirrhosis may prevent development of severe complications. Liver biopsy is the standard, but invasive and associated with morbidity. As noninvasive analysis of volatile organic compounds (VOCs) in breath was shown to distinguish cirrhotics from healthy controls, we aimed to identify a specific exhaled VOC profile to discriminate chronic liver disease patients with from those without cirrhosis. Methods: Breath samples were collected from 87 chronic liver disease patients without cirrhosis (CLD) and 46 with cirrhosis (CIR). A total array of 3718 exhaled VOCs was measured by gas chromatography time-of-flight mass spectrometry (GC-tof -MS). Partial Least Square Discriminant Analysis (PLS-DA) was performed to identify the discriminatory subset of compounds for classifying patients using a training set of 37 CIR and 37 CLD patients. Two independent validation sets (set 1 comprised 9 CIR and 9 CLD patients and set 2 the remaining 41 CLD patients) and a permutation test were used to validate the model. Results: A set of 22 VOCs was found to discriminate CIR and CLD patients with an overall correct prediction of 83.3% and 95.1% in validation set 1 and 2, respectively. Validation set 1 showed a sensitivity of 88%, specificity of 80% and negative and positive predictive values of 90% and 78%, respectively. Conclusions: Although further validation is required, this feasibility study showed that exhaled VOCs can accurately predict the presence of cirrhosis among CLD patients. Thereby, VOC analysis may be a potential non-invasive biomarker that can help to reduce the number of liver biopsies. P1021 COMPARISON OF DIAGNOSTIC VALUE OF ENHANCED LIVER FIBROSIS TEST WITH LIVER BIOPSY IN PATIENTS WITH AUTOIMMUNE HEPATITIS M.S. Gungoren1 , S.C. Efe2 , T. Kav2 , F. Akbiyik1,3 . 1 Department of Medical Biochemistry, 2 Department of Internal Medicine, Unit of Gastroenterology, Hacettepe University Faculty of Medicine, 3 Clinical Pathology and Clinical Chemistry Laboratories, Hacettepe University Hospitals, Ankara, Turkey E-mail: [email protected] Background and Aims: The enhanced liver fibrosis (ELF) test, a non-invasive diagnostic test panel consisting of hyaluronic acid (HA), amino-terminal propeptide of type III collagen (PIIINP), and tissue inhibitor of metalloproteinase 1 (TIMP-1) for the evaluation of liver fibrosis in chronic liver diseases. The aim of this study is to compare the performance of ELF test with biopsy in detection of liver fibrosis level in patients with autoimmune hepatitis.

Methods: Serum samples were collected from 46 patients with autoimmune hepatitis who have undergone liver biopsy. ELF panel parameters were measured with Siemens ADVIA Centaur XP immunoassay analyzer. ELF score was calculated with an algorithm using HA, PIIINP and TIMP-1 values. The METAVIR system was used to classify fibrosis stages in histopathological evaluation and cut-off for significant fibrosis was determined as ≥ F2. Diagnostic performances of ELF test and liver biopsy were evaluated with receiver operator characteristic (ROC) curve and area under ROC curve (AUROC) was calculated. Results: AUROC and best possible ELF test cut-off value for the prediction of significant fibrosis (≥ F2) were 0.88 and 8.84, respectively. Sensitivity, specificity, predictive values and likelihood ratios were as given in Table 1. Table 1.

Cut-off value AUROC Sensitivity Specificity Likelihood Ratio (+) Likelihood Ratio (−) Positive Predictive Value Negative Predictive Value Accuracy (%)

ELF score

95% Confidence interval

8.84 0.88 91.89 77.78 4.14 0.1 (10) 94.4 70 84.8

0.750–0.957 78.1–98.3 40.0–97.2 2.9–5.9 0.02–0.5 (20–50) 81.3–99.3 32.8–94.1

Conclusions: Our results suggest that ELF score can be used as a non-invasive marker for the assessment of liver fibrosis in autoimmune hepatitis patients and is especially useful in discriminating mild and significant fibrosis. P1022 QUANTIFICATION OF FIBROSIS IN CHRONIC HEPATITIS C: PERFORMANCE OF TRANSIENT ELASTOGRAPHY AND ACOUSTIC RADIATION FORCE IMPULSE USING COLLAGEN PROPORTIONATE AREA AS REFERENCE V. Calvaruso, V. Di Marco, F. Bronte, M.G. Bavetta, N. Alessi, C. Camma, ` A. Craxì. Gastroenterology and Hepatology, Di.Bi.M.I.S., University of Palermo, Palermo, Italy E-mail: [email protected] Background and Aims: Liver stiffness, measured by Transient Elastography (TE) or by Acoustic Radiation Force Impulse (ARFI), is proportional to liver fibrosis but also affected by necroinflammation. Since staging of fibrosis is semiquantitative and reported by classes, Collagen Proportionate Area (CPA), a continuous histological variable measuring collagen unaffected by necroinflammation, could correlate more proportionally with TE and ARFI values. Methods: Ninety-three consecutive patients with chronic hepatitis C (CHC) were evaluated for histological fibrosis (METAVIR score), CPA by Digital Image Analysis (DIA) and liver stiffness by TE and ARFI. Results: TE was unreliable in six patients (6.4%), while ARFI was feasible in all. By linear regression analysis TE correlated better than ARFI with CPA as a continuous variable (CPA–TE: R2 = 0.522, p < 0.001; CPA–ARFI: R2 = 0.454, p < 0.001) and with CPA quartiles (CPAq) (CPAq–TE: R2 = 0.434, p < 0.001; CPAq–ARFI: R2 = 0.334, p = 0.045). By comparison of ROC curves, the performance of TE in predicting CPA ≥10.5% (CPAq 3–4) and CPA >15.4% (CPAq 4) was significantly higher than that of ARFI (AUROC 0.884 vs 0.764, p = 0.006 and AUROC 0.863 vs 0.757, p < 0.001, respectively). At univariate analysis age, AST, ALT, GGT, platelets, TE, ARFI and Metavir-grade were related to CPAq 3–4. At multivariate logistic regression analysis, only TE (OR: 1.67, 95% CI: 1.13–2.48, p = 0.010) was independently related to CPAq 3–4. Similarly, AST, ALT, GGT, platelets, TE, ARFI and Metavir-grade were related to CPAq 4,


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POSTERS but by multivariate analysis only TE (OR: 1.39, 95% CI: 1.01–1.76, p = 0.040) was independently associated with CPAq 4 (>15.4%). The inflammation grade was marginally associated with CPA >15.4% (OR: 4.14, 95% CI: 0.98–17.56, p = 0.064). Conclusions: In patients with CHC, liver stiffness evaluated by TE and ARFI is proportionally related to CPA. TE is slightly more accurate than ARFI for the non-invasive staging of both significant and severe amount of liver fibrosis measured with CPA. P1023 OUR PRELIMINARY EXPERIENCE WITH ElastPQ® SHEAR WAVE ELASTOGRAPHY TECHNIQUE AND DOPPLER INDICES IN THE NON-INVASIVE ASSESSMENT OF LIVER FIBROSIS M. Garcovich, M.A. Zocco, L. Riccardi, E.B. Annicchiarico, M.E. Ainora, D. Roccarina, G. Caracciolo, A. Grieco, G.L. Rapaccini, M. Siciliano, M. Pompili, A. Gasbarrini. Catholic University of Sacred Hearth, Rome, Italy E-mail: [email protected] Background and Aims: Real-time shear wave elastography (RTE) is a novel non-invasive technique that assesses liver fibrosis by measuring liver stiffness (in kPa). The purpose of this study was to determine the efficacy and the feasibility for the assessment of hepatic fibrosis as compared with the histological grade in patients undergoing liver biopsy (LB). Methods: Consecutive patients scheduled for LB were studied by using the iU22 Philips ultrasound system with ElastPQ technique. In addition, Doppler indices at various sites, hepatic vein and portal venous blood velocity and flows were evaluated. The correlations between these quantitative parameters and the Metavir score were analyzed using Spearman correlation and ROC curve analyses were performed to calculate AUC for F > 2, F > 3, and F=4. Results: We enrolled 60 patients (39/21 males/females) who underwent LB for viral or non-viral chronic hepatitis (HCV 58%; NASH 30%). Liver stiffness measurements performed on the right lobe were reliable in almost all cases, while 15% of left lobe measurements were not obtainable/unreliable. Median kPa values were 4.43 (range 2.98–4.82) and 3.92 (2.51–6.73) for F0–F1, 7.65 (4.28–12.9) and 8.21 (5.43–12.3) for F2–F3, 15.12 (9.9–29.16) and 18.54 (9.31–31.34) for F4 in the right and left lobe, respectively. AUCs calculated for the right lobe were 0.90 (95% CI 0.84–0.92) for F > 2, 0.84 (95% CI 0.73–0.88) for F > 3 and 0.92 (95% CI 0.90–0.96) for F = 4. Adding Doppler indices to liver stiffness increased no further the diagnostic accuracy of RTE. Conclusions: RTE with ElastPQ appears to be a useful tool for noninvasive evaluation of fibrosis in patients with viral and non-viral chronic hepatitis, although these findings need to be confirmed in larger studies. P1024 PRESENCE OF HEPATIC STEATOSIS AS PER NON-INVASIVE BIOMARKERS OVERESTIMATES FIBROSIS STAGES BASED ON LIVER STIFFNESS MEASUREMENT (LSM) BY TRANSIENT ELASTOGRAPHY IN TYPE-2 DIABETIC (T2D) PATIENTS H. Perazzo1 , M. Munteanu2 , Y. Ngo2 , P. Lebray1 , E. Lukina1 , N. Seurat1 , F. Rutka3 , M. Couteau3 , S. Jacqueminet4 , D. Monneret3 , F. Imbert-Bismut3 , V. Ratziu1 , A. Hartemann-Huertier4 , C. Housset5 , T. Poynard1 , FLIP Consortium. 1 APHP UPMC Paris Liver Center, 2 BioPredictive, 3 Biochemistry, 4 Diabetology, Groupe Hospitalier Pitie Salpetriere, 5 INSERM UMR_S 938 & Saint Antoine Research Center, Paris, France E-mail: [email protected] Background and Aims: LSM and FibroTest (FT) are validated non-invasive methods to assess liver fibrosis. Necro-inflammatory activity influences LSM overestimating fibrosis. We aimed to evaluate the impact of hepatic steatosis on LSM in T2D patients. Methods: T2D patients without liver disease that had been screened for fibrosis with FT were reinvestigated using FT and S416

LSM after a 7-year median delay. Patients with minor fibrosis (FT 32% hepatocytes) was defined as SteatoTest ≥0.69, Fatty Liver Index (FLI) ≥60, Hepatic Steatosis Index (HSI) ≥36 or Controlled Attenuated Parameter (CAP) ≥283 dB/m. LSM applicability was defined as IQR/LSMratio 60% success rate and ≥10 valid measures. Results: 102 patients were pre-included [55% male, 62 years, BMI 27.6 kg/m2 ; ALT 23 (10–59), 35% insulin treated]. After exclusion of non-applicable LSM by both M and XL probes (7%), 95 patients were analyzed. Patients with supposed LSM overestimation compared to those without had higher; median [IQR]: BMI 32.0 [28.4–43.4] vs. 26.6 [24.4–35.3] kg/m2 ; waist circumference 109 [103–134] vs. 100 [93–119] cm; thoracic fold 24.4 [19.5–31.5] vs. 18.8 [16.5–27] mm; SteatoTest 0.64 [0.52–0.84] vs. 0.48 [0.32–0.83]; FLI 92 [61–99] vs. 60 [27–93]; and HSI 46 [40–53] vs. 38 [34–49], all p < 0.001. In a multivariate analysis [OR (95% CI)], severe steatosis as estimated by SteatoTest [5.2 (1.5–22.6); p = 0.01], FLI [5.8 (1.2–23.6); p = 0.03], HSI [6.4 (1.2–35.0); p = 0.03] or CAP [7.9 (1.4–45.0); p = 0.02] were associated with supposed LSM overestimation. Conclusions: The presence of hepatic steatosis in type 2 diabetic patients could overestimate liver fibrosis by liver stiffness measurement. P1025 UTILITY OF TRANSIENT ELASTOGRAPHY AND NON-INVASIVE FIBROSIS INDICES IN ASSESSING CHANGES IN FIBROSIS IN PATIENTS WITH CHRONIC HEPATITIS C E. Abdel Samea1 , W. Abdel Razek1 , N. Ehsan2 , M. Salama1 , I. Waked3 . 1 Hepatology, 2 Pathology, National Liver Institute, Shebeel El Kom, 3 Hepatology, National Liver Institute, Menoufiya University, Cairo, Egypt E-mail: [email protected] Background and Aims: Transient elastography (TE) and serum markers of fibrosis correlate with liver histology in the assessment of liver fibrosis in patients with HCV. Their utility in prospectively assessing changes in fibrosis is less clear. Aim: To compare the ability of TE and serum indices and scores to assess progress of liver fibrosis in HCV patients. Methods: 450 consecutive patients with HCV were followed for 2 years. TE and non-invasive tests of fibrosis (AST/ALT ratio, Forns’ index, Fibroindex, AST to platelet ratio index (APRI), Fib-4, Goteborg ¨ University cirrhosis index (GUCI), Lok index and Fibrosis index (FI)) were significantly correlated to initial biopsy, and were repeated after 24 months. Results: Patients were 39.2±10.2 years old and 67.3% were males. 269 (59.8%) patients received antiviral therapy, and 151 (56.1%) had sustained virological response (SVR). After 2 years, treated patients had significantly lower TE (−0.94±6.12 kPa vs. 0.95±3.48 kPa, p < 0.0001), Forns’ index (−0.1±1.1 vs. 0.1±0.8, p = 0.028) and Fibroindex (−0.04±0.4 vs. 0.03±0.3, p = 0.042). Other non-invasive tests did not show significant change between treated and untreated patients. Responders had decrease in TE (−2.82.8±4.4, p < 0.0001) and non-invasive tests, (Forns’ index: −0.5±0.8, Fibroindex −0.2±0.3, Lok index −2±13.7, FI −0.1±0.5) while in non-responders non-invasive tests and TE increased (TE +1.45±7.09 kPa). Conclusions: Fibrosis evaluated by TE and serum markers regressed after SVR, and progressed in non-responders. TE and Forn’s index could be useful in evaluating progress and change in fibrosis over time, and might be alternatives to serial biopsy.


POSTERS P1026 A NEW ECL-BASED ASSAY TO MEASURE SERUM CYTOKERATIN-18 IN PATIENTS WITH NONALCOHOLIC FATTY LIVER DISEASE E.P. Jeffries1 , J.A. Miller2 , J.A. White1 , C.K. Argo3 , S.H. Caldwell3 . 1 Wellstat Diagnostics, LLC, 2 Wellstat Biologics, LLC, Gaithersburg, MD, 3 GI/Hepatology, University of Virginia, Charlottesville, VA, United States E-mail: [email protected] Background and Aims: Nonalcoholic fatty liver disease (NAFLD) is a spectrum of disorders characterised by hepatic steatosis, which may be benign (NAFL) or which may progress via inflammation and fibrosis to nonalcoholic steatohepatitis (NASH) and then to cirrhosis and liver failure. Liver biopsy is the standard diagnostic approach for NAFL/NASH. However it has limitations due to sampling site variability, cost and procedure-related morbidity. Appropriate NAFLD-specific circulating biomarkers may enable diagnosis, staging and monitoring of NAFL/NASH with fewer biopsies. Circulating fragments of cytokeratin-18 (K18), a marker of hepatocyte death, have been shown in several studies to indicate the transition from benign fatty liver to NASH, with a risk of fibrosis, in patients with NAFLD. Our goal was to develop a highly specific serum K-18 assay to monitor hepatic disease severity in patients with different stages of NAFLD. Methods: An assay to measure K18 fragments in serum using electrochemiluminescence (ECL) technology was developed using proprietary K18 fragment-specific antibodies: one labelled with an ECL-active ruthenium chelate reporter and a second antibody bound to paramagnetic beads. Results: Thirty known NAFL and NASH patient serums with assigned NAS scores were evaluated. K18 fragment concentrations in biopsy-proven NASH were elevated compared to NAFL (1232 U/L and 345 U/L respectively). The results also demonstrated good correlation with the M30 antibody assay. Conclusions: An ECL-based assay has been developed for the quantitation of serum K18 fragments. This new diagnostic test may enable routine monitoring in both central laboratories and physician’s offices of disease severity and progression in patients with NAFLD. P1027 IS THE ELF-SCORE A VALID SUBSTITUTION FOR HVPG TO DETECT CLINICALLY SIGNIFICANT PORTAL HYPERTENSION (CSPH) NON-INVASIVELY? S. Hametner1 , A. Ferlitsch2 , A. Etschmaier2 , M. Ferlitsch2 , R. Schofl ¨ 1, A. Ziachehabi1 , D. Trubert-Exinger3 , A. Maieron1 . 1 Gastroenterology and Hepatology, KH d. Elisabethinen Linz, Linz, 2 Medical University of Vienna Department of Gastroenterology and Hepatology, Vienna, 3 Institute of Laboratory Medicine, LKH St. P¨ olten, St. P¨ olten, Austria E-mail: [email protected] Background and Aims: CSPH defined by HVPG ≥ 10 mm/Hg causes major complications. To improve survival, early diagnosis is crucial. vWF-Ag shows significant ability to diagnose CSPH and is a predictor for mortality. ELF-Score detects liver cirrhosis in most cases. Methods: 119 cirrhotic patients underwent HVPG measurement. Patients were categorized into 3 groups of different HVPG-ranges (0.2 >5 >3.25 >2.3 Fibro Q >2.6 >5.2 Forns Score >6.9 >5.0 Guci Index >0.26 >0.52 King Score >16.7 >10.1 Lok Index >0.5 PLF >2.98 >2.6

84 46 15 31 85 46 23 38 76 38 15 46 30 31 58

30 75 99 95 25 90 90 55 30 85 99 85 95 95 90

1.2 1.8 15 6.1 1.1 4.6 2.3 0.8 1.1 2.5 15 3.1 6.1 6.5 5.8

LR−

AUROC 95% CI

0.5 0.6 0.8 0.7 0.6 0.60 0.8 1.1 0.7 0.7 0.8 0.6 0.73 0.7 0.4

0.58 0.60 0.59

0.39–0.75 0.41–0.76 0.41–0.76

0.61

0.42–0.78

0.52

0.34–0.70

0.57

0.39–0.74

0.59

0.40–0.75

0.60 0.75

0.42–0.77 0.57–0.89

Values in boldface type are cut-offs indicated by literature.

Conclusions: Most of non-invasive serologic tests are not accurate in assessing fibrosis stage and identifying cirrhotics after an SVR. Histological evaluation through liver biopsy still remains the gold standard in these patients. P1033 CARBONYLATED PROTEINS AND GLUTATHIONE LEVELS AS IMPORTANT OXIDATIVE BIOMARKERS IN ALCOHOLIC LIVER DISEASE ´ M. Galicia1 , D. Rosique Oramas1 , T. Alvarez Torres1 , L. Very 2 ´ Hernandez ´ , Pineda1 , K. Medina Avila1 , L. Raya Soto1 , J.L. Perez ´ Reyes1 . 1 Unidad de Medicina D. Kershenobich1 , G. Gutierrez Experimental, Facultad de Medicina Universidad Nacional Aut´ onoma de Mexico, 2 Clinica de H´ıgado, Hospital General de Mexico, Mexico City, Mexico E-mail: [email protected] Background and Aims: The free radicals and the oxidative stress plays an important role in the pathogenesis of alcoholic liver disease (ALD); this has been demonstrated by several markers of oxidative damage and the imbalance of antioxidant defenses in animal models, but little is known about this process in the human. Aim: To evaluate the role of oxidative stress through the quantification of carbonylated proteins and the levels of glutathione in patient with ALD. Methods: Two groups of patients (n = 44) were formed and included in cross-sectional study. Group 1, control group consisting by subjects with an ethanol consumption ≤10 g/day and AUDIT ≤8.

Table 1. Results of clinical and biochemical parameters determinate in the study Parameter

Control

Patients with ALD Child–Pugh A

Child–Pugh B

Gender (F/M) Age

10/34 38.1±1.4

0/16 45.9±2.7a

1/16 47.2±1.9a

0/11 53.2±3.5a

Body mass index (kg/m2 ) Consumption OH (g/day)

28.2±0.4 1.5±0.3

Carbonylated proteins (nmol/mg protein)

0.02±0.003

29±1.2 333.1±54.9a 0.06±0.01d

29±1.3 362.6±73.6a 0.07±0.02d

26.3±0.8 242.1±30.6a 0.05±0.01abc

GSH (mM) GSSG (mM)

540.7±17.7 133.4±16.5

GSH/GSSG

3.7±0.5

733.4±41.2ad 227.6±73.8 -5.6±3.4acd

616.9±32.2 175.2±28.5 2.7±0.5b

551.6±40.2b 347.3±91a 1.76±1.1b

Child–Pugh C

Values are presented as mean±S.E. a P ≤ 0.05 versus Control; b P ≤ 0.05 versus Child Pugh A; c P ≤ 0.05 versus Child–Pugh B; d P ≤ 0.05 versus Child–Pugh C.

P1034 ESTIMATED PREVALENCE OF HCV CIRRHOSIS IN THE CHRONIC HEPATITIS COHORT STUDY (CHECS) S.C. Gordon1 , L. Lamerato2 , L.B. Rupp3 , S.D. Holmberg4 , A.C. Moorman4 , P.R. Spradling4 , E. Teshale4 , V. Vijayadeva5 , J.A. Boscarino6 , M.A. Schmidt7 , D. Nerenz3 , N. Oja-Tebbe2 , M. Lu2 , for the CHeCS Investigators. 1 Gastroenterology and Hepatology, 2 Public Health Sciences, 3 Center for Health Policy and Health Services Research, Henry Ford Health System, Detroit, MI, 4 Division of Viral Hepatitis, Centers for Disease Control and Prevention, Atlanta, GA, 5 Center for Health Research, Kaiser Permante Hawaii, Honolulu, HI, 6 Center for Health Research, Geisinger Clinic, Danville, PA, 7 Center for Health Research, Kaiser Permanente Northwest, Portland, OR, United States E-mail: [email protected] Background and Aims: The prevalence of cirrhosis among patients with chronic hepatitis C (CHC) may be underestimated. We used four different methods to estimate the prevalence of cirrhosis among CHC patients enrolled in the Chronic Hepatitis Cohort Study (CHeCS). Methods: Cirrhosis was identified via four methods: 1. liver biopsy report, 2. presence of a diagnosis code for cirrhosis, 3. presence of a diagnosis or procedure code indicating ESLD, and 4. FIB-4 score ≥5.88, a cut-off value previously validated as predictive of cirrhosis in this cohort. We excluded patients who were coinfected with hepatitis B, liver transplant recipients, and those who had previously achieved SVR. We compared the extent to which cirrhosis prevalence was identified by each method. Results: Of the 9,783 CHC patients included in the study, 4,221 (43%) had at least one biopsy report during follow-up, and of those, 634 (15%) had a recent biopsy 10 valid measurements, SR > 60% and IQR < 30%. Liver fibrosis was staged by METAVIR in HCV RNA positive patients. Intra-class correlation coefficients and AUROC were used for inter-observer agreement and diagnostic accuracy. Results: LS and SS resulted highly reproducible (0.98, 95% CI 0.95–1.0 and 0.86, 95% CI 0.80–0.91, respectively). At multivariate analysis, major determinants of LS were ALT and PT ratio (p < 0.0001), and for SS splenic volume (p < 0.0001). In TM and TI, LS had a good diagnostic power in ruling out severe fibrosis/cirrhosis (AUROC 0.93, sensitivity 100%, specificity 70%, LR+ 4; LR− 0.01) in the subset of patients who underwent liver biopsy. Conclusions: LS and SS determination by TE are highly reproducible tools in patients with haemoglobinopathies. The two parameters resulted significantly higher in patients than in controls and not influenced by iron overload. LS was highly accurate in excluding severe fibrosis/cirrhosis. S420

P1036 ASSESSMENT OF LIVER FIBROSIS IN PATIENTS WITH AUTOIMMUNE LIVER DISEASES USING ARFI ELASTOGRAPHY D. Attia1,2 , A. Dettmer1 , H. Bantel1 , M.P. Manns1 , M. Gebel1 , A. Potthoff1 . 1 Gastroenterology, Hepatology and Endocrinology, Hannover Medical School, Hannover, Germany; 2 Beni-Suef University, Beni-Suef, Egypt E-mail: [email protected] Background and Aims: Transient elastography (TE) is considered as one of the best non-invasive methods staging liver fibrosis in primary biliary cirrhosis. Only few data are present about the diagnostic accuracy of ARFI elastography in patients with autoimmune liver diseases. Therefore, the aim of our study was to evaluate prospectively the diagnostic accuracy of liver stiffness measured by ARFI elastography in comparison to liver biopsy and TE in patients with autoimmune liver diseases. Methods: A total of 112 patients with autoimmune hepatitis (AIH, n = 62) and primary biliary cirrhosis (PBC, n = 50) (82 females, 30 males; mean age 53±14 y) were enrolled in the study. All patients received ARFI elastography. Results were compared with TE and liver biopsies in order to evaluate the diagnostic accuracy. Results: The diagnostic accuracy of ARFI detecting liver fibrosis ≥ F3 and ≥ F4 in patients with AIH were 0.85 and 0.98, respectively, in comparison to liver biopsy and 0.93 and 0.96 in comparison to TE with cut off values of 1.34 m/s, 1.56 m/s and 1.47 m/s, 1.56 m/s, respectively. In patients with PBC, the diagnostic accuracy detecting liver fibrosis ≥ F3 and ≥ F4 were 0.96 and 0.88, respectively, in comparison to liver biopsy and 0.92 and 0.97, respectively, in comparison to TE with cut off values of 1.64 m/s, 2.50 m/s and 1.48 m/s, 2.46 m/s, respectively. Conclusions: ARFI elastography is able to diagnose advanced liver fibrosis and liver cirrhosis in patients with autoimmune liver diseases with an excellent diagnostic accuracy. However, cut-off values were different from those with viral hepatitis. P1037 NON-INVASIVE MODALITIES FOR THE DIAGNOSIS OF ADVANCED FIBROSIS AND CIRRHOSIS IN CHRONIC HEPATITIS B OR C R.W.-J. Jeng1,2 , P.-K. Tsay3 , W.-T. Chen1,2 , C.-H. Huang1,2 , S.-M. Lin1,2 , D.-I. Tai1,2 . 1 Department of Gastroenterology and Hepatology, Division of Hepatology, Chang Gung Memorial Hospital, 2 Chang Gung University College of Medicine, 3 Graduate Institute of Biomedical Science, Chang Gung University, Taipei, Taiwan E-mail: [email protected] Background and Aims: The diagnosis of advanced fibrosis and cirrhosis is important in patients with chronic hepatitis B or C, especially in treatment decision making and prognosis. We compare non-invasive diagnostic modalities for the diagnosis of advanced fibrosis or cirrhosis. Methods: Consecutive CHB or CHC infected patients with liver biopsy were prospectively recruited. Hemogram, biochemistry, conventional ultrasonography and acoustic radiation force impulse elastography (ARFI) were measured on the same day of biopsy. Advanced fibrosis was defined as ISHAK score ≥4 and cirrhosis as ≥5. ARFI was done in two locations of R lobe liver with 10 measurements separately. Logistic regression and receiver operating characteristic (ROC) curve analyses were performed to compare patients groups with ISHAK score ≥4 or ≥5. Results: A total number of 212 patients (Age: mean: 50.3±11.2; 72.6% male; 60% CHB, 40% CHC) were recruited; 35.8% were advanced fibrosis and 28.8% were cirrhosis. By multivariate logistic regression, spleen index (OR: 1.09; 95% CI: 1.034–1.157; p = 0.002), conventional ultrasonography (OR: 2.43; 95% CI: 1.78– 3.32; P = 0.000) and ARFI (OR: 3.51; 95% CI: 1.59–7.75; p = 0.002) are predictive of advanced fibrosis. For cirrhosis, spleen index (OR: 1.07; 95% CI: 1.01–1.13; p = 0.02), conventional ultrasonography (OR:


POSTERS 2.59; 95% 1.89–3.53; P = 0.000) and ARFI (OR: 4.2; 95% CI: 2.0–8.73; p = 0.000) were independent predictors. The AUROCs are shown in Table 1. The optimal ARFI cutoff value for advanced fibrosis was ≥1.33 m/s and ≥1.4 m/s for cirrhosis.

of Hepatology GH Pitié-Salpêtrière, 4 Hepatology Research Unit, BioPredictive, 5 ANRS, 6 BioPredictive, 7 Biochemistry Department GH Pitié-Salpêtrière, APHP UPMC Paris Liver Center, Paris, France E-mail: [email protected]

Table 1. AUROC for advanced fibrosis and cirrhosis

Background and Aims: Applicability of non-invasive methods, liver stiffness measurements (LSM) by Fibroscan and Fibrotest, are defined after excluding failures and unreliable results (Castera 2010, Poynard 2010). RT-SWE by Aixplorer is a new 3D-elastography coupled with a B-mode imaging. Aims: To better identify applicability criteria for SWE in terms of positioning and acquisition procedure. Methods: We used standard-AUROCs and the strength of concordance with validated methods, LSM and Fibrotest. Each patient had 3-SWE regions of interest (ROI) studied [right lobe (RL)] with 3 measures within each: central-zone (Q-boxC), zone displayed in blue-color (Q-boxB), and in red-color (Q-boxR). Results: 189 prospective-patients were pre-included with App-FT; 166 (87.8%) had concomitant App-LSM: 57% males, age 53 yrs, BMI 25 kg/m2 , ALT 40 IU/L; 30% advanced fibrosis (AF) as per Fibrotest. Q-boxC and the mean of three C-measurements correlated with Fibrotest (p < 0.05). Q-boxB correlated with Fibrotest (p < 0.05) unlike Q-boxR (p = NS). Taking Fibrotest as standard, the best AUROCs were obtained for Q-boxC (0.73) and Q-boxB (0.74), but not different from the mean of 3 Q-boxC (0.71) and Q-boxB (0.69). Using the mean of 3 Q-boxC (-B) does not improve diagnosis provided by Qbox-C (-B) alone (p = NS). SWE with a ratio SD/meanSWE >60% versus 7.1 kPa. We evaluated the diagnostic power and accuracy of TE measurements according to these new quality criteria (accurate = very reliable + reliable) for non-invasive assessment of liver fibrosis (liver biopsy) and portal hypertension (HVPG). Methods: Patients undergoing TE, HVPG measurement and liver biopsy within 3 months at a tertiary care were retrospectively identified. Results: Among 278 patients (48.7±13.1 years, 74.7% male, 75.7% viral etiology, 57% F3/F4), traditional TE quality criteria identified 71.6% reliable measurements, while new criteria yielded in 83.2% accurate LS measurements (23.1% very reliable, 60.1% reliable). Reliable TE values according to traditional or new criteria were all significantly and similarly strong correlated with fibrosis stage (R = 0.648 vs. R = 0.636) and HVPG (R = 0.836 vs. R = 0.846). The accuracy for diagnosing liver cirrhosis (F4, cut-off: 14.5 kPa) was 76.5% and 75.0% for traditional and new TE criteria, respectively. The positive (PPV) and negative (NPV) values for new criteria at the 14.5 kPa cut-off were 83% and 70%. For predicting HVPG ≥10 mmHg (cut-off: 16.1 kPa), the accuracies were 88.9% and 89.8% using traditional or new criteria, respectively. Both criteria resulted in AUCs for diagnosis of HVPG ≥10 mmHg of over 0.95 with a PPV and NPV of 76% and 97%, respectively. Conclusions: Applying new quality criteria for TE measurements significantly increases the number of valid TE measurements without affecting accuracy of TE for diagnosis of liver cirrhosis and portal hypertension. P1039 APPLICABILITY (APP) CRITERIA FOR REAL-TIME SHEAR WAVE ELASTOGRAPHY (RT-SWE) BY AIXPLORER COMPARED TO TRANSIENT ELASTOGRAPHY (TE) BY FIBROSCAN AND TO FIBROTEST N. Seurat1 , H. Perazzo2 , E. Luckina2 , L. Royer2 , M. Munteanu3 , Y. Ngo4 , L. Fedchuk5 , O. Deckmyn6 , D. Monneret7 , F. ImbertBismut7 , V. Ratziu1 , T. Poynard1 . 1 Department of Hepatology GH Pitié-Salpêtrière, 2 APHP UPMC Paris Liver Center, 3 Department

7C. VIRAL HEPATITIS: HEPATITIS B & D – CLINICAL (THERAPY, NEW COMPOUNDS, RESISTANCE) P1040 TENOFOVIR FOR THE PROPHYLAXIS OF HBV REACTIVATION IN ANTI-HBC-POSITIVE PATIENTS WITH HEMATOLOGIC MALIGNANCIES TREATED WITH RITUXIMAB: PRELIMINARY RESULTS OF A RANDOMIZED STUDY (PREBLIN STUDY) M. Buti1 , R. Morillas2 , M.L. Manzano3 , M. Garc´ıa-Retortillo4 , 5 8 , L. Mart´ın6 , M. Prieto7 , E. Suarez ´ , F. Gea9 , M.L. Gutierrez ´ 12 13 ´ Simon ´ , M. Gomez ´ , M.A. ´ 14 , M. Rodr´ıguez10 , J.M. Zozaya11 , J. Lopez 15 16 1 1 L.E. Morano , A. Pardo , R. Esteban . Hepatology, Vall d’Hebron Hospital, Barcelona, 2 Hepatology, Germans Trias i Pujol Hospital, Badalona, 3 Hepatology, October 12 Hospital, Madrid, 4 Hepatology, Del on, Alcorc´ on, Mar Hospital, Barcelona, 5 Hospital Foundation Alcorc´ an, 7 Hepatology, Madrid, 6 Hepatology, Donostia Hospital, San Sebasti´ La Fe Hospital, Valencia, 8 Hepatology, Valme Hospital, Sevilla, 9 Hepatology, La Paz Hospital, Madrid, 10 Hepatology, Central Hospital of Asturias, Oviedo, 11 Gastroenterology, Navarra Hospital, Pamplona, 12 Hematology, Ram´ on y Cajal Hospital, Madrid, 13 Gastroenterology, Getafe Hospital, Getafe, Madrid, 14 Hepatology, Lozano Blesa Hospital, Zaragoza, 15 Infectious Diseases, Do Meixoeiro Hospital, Vigo, 16 Gastroenterology, Joan XXIII Hospital, Tarragona, Spain E-mail: [email protected] Background and Aims: Patients positive for HBsAg or anti-HBc are at risk of severe HBV reactivation due to immunosuppressive treatment with rituximab (RTX). Lamivudine prophylaxis reduces


S421

POSTERS but does not eliminate this risk. It is important to assess the safety and efficacy of more potent antivirals, such as tenofovir (TDF). Methods: Randomized, prospective, open-label, multicenter, parallel-group clinical trial. HBsAg-negative, anti-HBc-positive patients with undetectable HBV-DNA were randomized before starting RTX to receive TDF or to 18 months of observation (tests every 2 months). HBV reactivation defined as HBV-DNA elevation ≥ 1 log10 IU/mL above baseline and/or HBsAg reappearance. Results: To date, 69 patients assessed and 50 enrolled. An interim analysis including first 6 months of follow-up was performed. During this period, 4 subjects withdrawn, 3 for blood diseaserelated death and 1 for adverse event. Interim efficacy analysis included 25 patients, 66% male, median of 70 years (31–86), with non-Hodgkin lymphoma (62%) and chronic lymphocytic leukemia (29%). Three patients had detectable HBV-DNA at baseline and received open-label TDF. Remaining 22 patients were randomized, 11 to each treatment group. Number of HBV reactivations in that period was 0/14 (0%) with TDF and 2/11 (18%; P=.2) with observation. Reactivations were identified at 4-month visit (HBV-DNA elevation without HBsAg seroreversion) and rescued with TDF, showing undetectable HBV-DNA at 6-month visit. Results will be updated at the presentation time, including a larger sample size and a longer evaluation time. Conclusions: This interim analysis shows that 18% of anti-HBcpositive subjects treated with RTX developed HBV reactivation. Pre-emptive TDF prevented HBV reactivation in all cases. P1041 TENOFOVIR MONOTHERAPY VERSUS LAMIVUDINE PLUS ADEFOVIR IN LAMIVUDINE-FAILURE PATIENTS RESCUED WITH LAMIVUDINE PLUS ADEFOVIR COMBINATION: INTERIM ANALYSIS OF THE TENOSIMP-B CLINICAL TRIAL F. Jorquera1 , J.M. Pascasio2 , E. Fraga3 , J. Fuentes4 , M. Prieto5 , G. Sanchez-Antol´ ´ ın6 , J.L. Calleja7 , E. Molina8 , S. Tome´ 9 , L. Bonet10 , ´ Blanco11 , M.L. Garc´ıa-Buey12 , J. Salmeron ´ 13 , J.A. Pons14 , M.A. J.M. Gonzalez ´ 15 , M. Rodr´ıguez16 . 1 Gastroenterology, Hospital Complex of Le´ on, Le´ on, 2 Hepatology, Virgen del Rocio Hospital, Sevilla, 3 Reina Sof´ıa University Hospital, C´ ordoba, 4 Hepatology, Miguel Servet Hospital, Zaragoza, 5 Hepatology, La Fe Hospital, Valencia, 6 Hepatology, Rio Hortega Hospital, Valladolid, 7 Hepatology, Puerta de Hierro Hospital, Majadahonda, Madrid, 8 Hepatology, 9 Infectious Diseases, University Hospital Complex of Santiago, Santiago de Compostela, 10 Gastroenterology, Son Espases Hospital, Palma de Mallorca, 11 Hepatology, Gregorio Mara˜ no ń Hospital, 12 Gastroenterology, La Princesa Hospital, Madrid, 13 Hepatology, San Cecilio Hospital, Granada, 14 Gastroenterology, Virgen Arrixaca Hospital, El Palmar, Murcia, 15 Hepatology, Clinic Hospital, Valladolid, 16 Hepatology, Central Hospital of Asturias, Oviedo, Spain E-mail: [email protected] Background and Aims: To date, there are no published reports on clinical trials comparing the efficacy of the lamivudine plus adefovir (LAM+ADV) combination versus tenofovir (TDF) monotherapy in LAM-failure patients with undetectable viremia. Methods: Prospective, randomized, open-label, multicenter, parallel-group, non-inferiority study at 14 Spanish sites. The primary endpoint was the percentage of patients with undetectable HBV DNA during 1 year of follow-up. Renal function and treatment adherence were also assessed. Clinical, chemistry, and virologic follow-up were performed every 3 months. Results: 53 patients were randomized: 26 to TDF and 27 to LAM+ADV. Most (81%) were men; the median age was 57 years (32–84). Except for 1 subject, all were HBeAg-negative at baseline. Both treatment groups were similar in age and gender. As of September 2013, 45 patients had completed 1 year of follow-up; 4 of these dropped out (3 receiving LAM+ADV and 1 receiving TDF). No detectable HBV DNA episodes were observed. ALT levels S422

remained stable and below 30 U/L in both groups. At baseline, both treatments presented an average phosphatemia of 2.8 mg/dL, blood creatinine of 0.9 mg/dL, and glomerular filtration rate (GFR, by MDRD) of 90 and 96 mL/min/1.73 m2 for TDF and LAM+ADV, respectively (NS). Changes from baseline in phosphatemia and GFR are shown in the table. Treatment

Renal safety parameter, change from baseline Phosphatemia (mg/dL)

LAM+ADV TDF P*

Glomerular filtration rate (mL/min/1.73 m2 )

6 months

12 months

6 months

12 months

+0.10 (n = 19) −0.01 (n = 17) 0.42

+0.06 (n = 13) +0.08 (n = 15) 0.96

−3.55 (n = 21) −3.28 (n = 20) 0.79

−7.01 (n = 14) −0.92 (n = 18) 0.29

*According to Mann-Whitney test for TDF vs LAM+ADV.

Patients identified as highly adherent were 89% and 86% at 12 months for TDF and LAM+ADV (NS), respectively. Conclusions: This interim analysis showed that treatment simplification with TDF was not inferior to LAM+ADV in patients with prior failure to LAM. P1042 EFFICACY OF ENTECAVIR MONOTHERAPY VERSUS ENTECAVIR PLUS ADEFOVIR COMBINATION THERAPY AS RESCUE THERAPY FOR rt181 MUTANTS OF HEPATITIS B VIRUS: 48 WEEKS RESULTS M.J. Oh, H.J. Lee, K.H. Kim, K.O. Kim, S.H. Lee, B.I. Jang, T.N. Kim. Department of Internal Medicine, Yeungnam University College of Medicine, Daegu, Korea, Republic of E-mail: [email protected] Background and Aims: Mutation at locus 181 of reverse transcriptase (rt) of hepatitis B virus (HBV) may predict HBV DNA persistence and incomplete therapeutic response, and is known as cross-resistance against other nucleos(t)ide analogues (NUCs), except entecavir. We aimed to investigate long-term efficacy of NUCs as rescue therapy for rt181 mutants of HBV. Methods: Between April 2008 and October 2011, among 797 patients who performed sequencing analysis of HBV polymerase due to virological breakthrough after NUCs therapy, a total of 95 patients who received entecavir (1.0 mg) monotherapy (n = 28) or entecavir (1.0 mg) plus adefovir therapy (n = 67) for rt181 mutants over 48 weeks were selected. The subjects were retrospectively analyzed for virological and biochemical responses at 48 weeks. Results: Median age of enrolled patients was 46.0 years. The mean duration of NUCs therapy before detection of rt181 mutation was 270.8±153.4 weeks. Virological responses after rescue therapy of 48 weeks showed complete virological response (17.9 vs. 6.0%), partial virological response (46.4% vs. 70.1%), non-response (32.1% vs. 20.9%), and virological breakthrough (3.6% vs. 3.0%), respectively. Biochemical response in the both groups was 42.9% vs. 25.4%. No significant difference was observed in virological, and biochemical response, respectively (p = 0.098, p = 0.141). The rate of HBeAg loss or seroconversion did not significantly differ between the two groups (37.5% vs. 21.3%; p = 0.169). The mean reduction rate of HBV DNA real-time PCR of each group at 48 weeks was not different statistically (4.2 vs. 4.5 log10 IU/mL; p = 0.595). Conclusions: Our study demonstrated that long-term efficacy of entecavir monotherapy as rescue therapy against rt181 mutants was clinically comparable to that of combination therapy of entecavir plus adefovir.


POSTERS P1043 REAL-LIFE PERSISTENCE AND ADHERENCE IN ETV TREATED CHRONIC HEPATITIS B PATIENTS: RESULTS OF A GERMAN PROSPECTIVE MULTICENTER OBSERVATIONAL STUDY J. Petersen1 , T. Wilke2 , S. Mauss3 , R. Heyne4 , C. Herold5 , M. Wiese6 , K. Boeker7 , T. Pichl8 , D. Hueppe9 . 1 ifi – Institut f¨ ur Interdisziplin¨ are Medizin, Hamburg, 2 IPAM Wismar, Wismar, 3 Medizinisches Versorgungszentrum, Duesseldorf, 4 Leberzentrum ur Checkpoint, Berlin, 5 Internisten am Ring, Nuernberg, 6 Gesellschaft f¨ Klinische Studien, Leipzig, 7 Leberpraxis Hannover, Hannover, 8 BristolMyers Squibb GmbH & Co. KGaA, Munich, 9 Gastroenterologische Gemeinschaftspraxis Herne, Herne, Germany E-mail: petersen@ifi-medizin.de Background and Aims: Entecavir (ETV) is a very effective and safe treatment option in chronic hepatitis B (CHB) patients. However, some patients show only a partial virologic response. The most reasonable explanation is non-persistence/non-adherence. Persistence and adherence measurement is underrepresented in clinical trials. Therefore, the extent of non-persistence (NP), nonadherence (NA) and clinical outcomes of any NP/NA were measured in ETV-treated patients in a real-life study. Methods: In a prospective observational multicenter study, persistence and adherence were measured based on documented ETV prescriptions within a 12 month period. A patient was defined as NP if he missed all treatment doses in >30 subsequent days. Adherence was measured based on medication possession ratio (MPR) between first and last documented prescription (NA if MPR < 80%). Proportion of patients reaching undetectable HBV-DNA levels (30 subsequent days (NP group), 30% of patients in the NP group (n = 9) and none of the persistent patients were classified as non-adherent. Mean MPR for all patients was 95.3%. 84.4% of persistent/adherent patients and 63.0% of patients in the NP group reached undetectable HBV-DNA levels (p = 0.022). So far no significantly correlating factors for NP/NA could be detected. Table 1. Selected baseline characteristics and results Baseline characteristics (N = 112) Age, mean (years) 45.8 Gender (% female) 42.0 eAg positive patients (%) 29.5 4.7 HBV-DNA, mean (IU/ml log10 ) Proportion of patients (%) with undetectable HBV-DNA (