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tent. Results: Collagen deposition in response to BDL increased over time as assessed by Hyp analysis and Sirius red staining. Compared to sham controls, liver ΔR1 increased significantly in BDL rats at 10 and 18 days (p=0.04 and p<0.01, respec-tively). Liver ΔR1 also correlated with ex vivo Gd concentration (R=0.65). Rapamycin inhibited biliary fibrosis progression as measured by Sirius red staining (p<0.01). Consistently, a sig-nificant decrease in ΔR1 was observed in BDL rats that received rapamycin (p=0.01). Interestingly, marked heterogeneity was observed by Sirius red staining between the left and right lobes of a “non-responder” rapamycin-treated rat, and this difference was detected on the ΔR1 maps. Conclusion. Quantitative 3D ΔR1 maps can be used to monitor progression of biliary fibro-sis throughout the entire liver, and ΔR1 is sensitive to changes in liver fibrosis in response to rapamycin treatment. EP-3533 should be examined for its ability to image liver fibrosis in humans.Disclosures:Kenneth K. Tanabe - Patent Held/Filed: EGF SNP and risk for HCC, EGFR inhibi-tion and HCC, gene signature for prognosis in cirrhosisPeter Caravan - Grant/Research Support: Sanofi; Stock Shareholder: Collagen MedicalThe following people have nothing to disclose: Bryan C. Fuchs, Christian T. Farrar, Danielle K. DePeralta, Helen Day, Boris Keil, Gregory Y. Lauwers, Lan Wei, Arun Subramaniam

19The use of simeprevir and sofosbuvir to treat HCV G1 in the Liver Transplant Setting: The experience in 3 US CentersBashar Aqel1, Surakit Pungpapong2, K Tuesday Werner1, Amy E. Chervenak1, Jorge Rakela1, Kymberly D. Watt3, Michael D. Leise3, Jennifer L. Murphy2, Tanisha M. Henry2, Kristen Ryland2, Andrew Keaveny2, Hugo E. Vargas1; 1Hepatology, Mayo Clinic Arizona, Phoenix, AZ; 2Transplantation, Mayo Clinic Florida, Jacksonville, FL; 3Gastroenterology/Hepatology, Mayo Clinic Minnesota, Roch-ester, MNResults from a phase II study (COSMOS) suggested that HCV G1 infection can be treated effectively with a combination of sofosbuvir (SOF) and simeprevir (SMV) with or without rib-avirin (RBV) for 12 weeks. The regimen is suitable for IFN ineligible patients and those who have failed prior treatment with advanced fibrosis. Objective: To report the experience of treating patients who were IFN ineligible/prior treatment failures with SOF/ SMV combination in 3 U.S. liver transplant (LT) centers. Results: We identified 127 patients with G1 dis-ease who required this IFN-free treatment. To date 91 (71.7%) have initiated treatment. Of the 91 patients, 60.4% were male, 89% were Caucasian. 55% had failed prior treatment, 15% relapsed and 30% were treatment naïve. 82.4% had cirrhosis and of those 36% are listed for LT. (median MELD was 9, range 6-22). To date, 19 have completed 12 weeks of therapy treat-ment; 70% were HCV RNA negative at week 4; all were HCV RNA negative at week 12. So far, no treatment relapses have occurred in these patients; 1 patient received a LT 6 weeks after she became virus negative and remains virus negative 2 weeks post-LT. No serious adverse events or episodes of hepatic decompensation have been observed. Four patients have reported self-limited vertigo on treatment. Conclusion: SMV/SOF combination has been well tolerated in our difficult to treat population of patients a majority of whom are cirrhotic, and who are ineligible, previously intolerant or non-responsive to IFN-based therapy. No episodes of hepatic decompensation have been documented with this regimen to date. A major bar-rier to initiating SMV/SOF combination treatment is the slow approval process, even in patients with advanced liver dis-

ease. SVR 12 data will be presented as it becomes available that will allow better characterization of the benefit of SMV/SOF therapy in cirrhotic patients.Disclosures:Surakit Pungpapong - Grant/Research Support: BMS, GileadHugo E. Vargas - Advisory Committees or Review Panels: Eisai; Grant/Research Support: Merck, Gilead, Idenix, Novartis, Vertex, Janssen, Bristol Myers, Ikaria, AbbVieThe following people have nothing to disclose: Bashar Aqel, K Tuesday Werner, Amy E. Chervenak, Jorge Rakela, Kymberly D. Watt, Michael D. Leise, Jennifer L. Murphy, Tanisha M. Henry, Kristen Ryland, Andrew Keaveny

20Prevention of Hepatitis C Virus (HCV) Recurrence with Peri-Transplant Hepatitis C Immune Globulin Combined with Pre-Transplant (Pre-LT) Antiviral Therapy (AVT)Norah Terrault1, Sanjaya K. Satapathy2, George Therapondos3, Roshan Shrestha4, Elizabeth C. Verna5, Jeffrey Campsen6, James Spivey7, Jens Rosenau8, Thomas D. Schiano9, Kalyan R. Bhamidi-marri10, Sher Linda11, John M. Vierling12, Lewis W. Teperman13, Gerond Lake-Bakaar14, Jacqueline G. O’Leary15, Laura M. Kulik16, Fredric D. Gordon17, Daniel Maluf18, Shailesh Chavan19, Chris-topher J. Dougherty19; 1University of California, San Francisco, CA; 2University of Tennessee Health Sciences Center, Memphis, TN; 3Ochsner Clinic Foundation, New Orleans, LA; 4Piedmont Transplant Institute, atlanta, GA; 5Columbia University, New York, NY; 6University of Utah, Salt Lake City, UT; 7Emory University, Atlanta, GA; 8University of Kentucky, Lexington, KY; 9Mount Sinai Medical Center, New York, NY; 10University of Miami, Miami, FL; 11University of Southern California, Los Angeles, CA; 12Baylor College of Medicine, Houston, TX; 13New York University, New York, NY; 14Beth Israel Deaconess Medical Center, Boston, CT; 15Baylor University Medical Center, Dallas, TX; 16Northwestern University Feinberg School of Medicine, Chicago, IL; 17Lahey Hos-pital and Medical Center, Burlington, MA; 18University of Virginia Health System, Charlottesville, VA; 19BIotest Pharmaceuticals, Boca Raton, FLBackground and Aims: Safer HCV AVTs are available to treat wait-listed patients to prevent post-LT HCV recurrence, but such therapies are not uniformly effective and the optimal duration of pre-LT AVT unknown. We evaluated the safety and efficacy of Biotest-HCIG, a human hepatitis C immune globulin to pre-vent HCV recurrence by neutralizing remaining HCV reservoirs in patients on pre-LT HCV AVT at the time of LT. Methods: In this phase 3, open-label randomized study, wait-listed patients with chronic HCV infection (all genotypes) treated with any AVT and who achieved HCV RNA <100 IU/ml prior to LT were eligible. In total, 84 patients will be randomized 1:1:1 to Biotest-HCIG (200 mg/kg or 300 mg/kg given on the day of LT and for 10 weeks post-LT) or observation. The primary endpoint is post-LT sustained virologic response (pTVR), defined as HCV RNA <43 IU/ml at 12 wks post-LT treatment. Post-transplant immunosup-pression is site-specific. Results: To date, 17 subjects (all male, median age 59 yrs, 100% genotype 1, 94% with hepatocel-lular carcinoma, 12% with living donors) have undergone LT. Pre-LT AVT was telaprevir/peginterferon/ribavirin (RBV) (12%), sofosbuvir/RBV (76%) or sofosbuvir/simeprevir (12%) given for a median of 51 days (range 14-164 days) pre-LT with all patients achieving HCV RNA <43 IU/mL pre-LT (71% also undetectable). With median post-LT follow-up of 8 wks, post-LT HCV recurrence has been documented in 2 patients - at wk 2 (control) and wk 3 (200 mg Biotest-HCIG) post-LT. Overall, 11/12 (92%) of Biotest-HCIG-treated patients have maintained undetectable HCV RNA compared to 4/5 (80%) of controls (Table). Among 4 patients who were viremic at the time of LT and randomized to Biotest-HCIG, all have undetectable HCV

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RNA at median 9 wks follow-up. Biotest-HCIG-related side effects were infrequent and there were no discontinuations due to adverse events. Conclusion: Biotest-HCIG is safe and well-tol-erated. To date, HCV recurrence rates in patients on pre-LT AVT are lower in Biotest-HCIG-treated patients compared with con-trols (8% vs 20%) and all patients viremic at LT who received Biotest-HCIG have undetectable HCV RNA. These preliminary results suggest Biotest-HCIG may be beneficial as an adjuvant therapy for HCV patients on AVT undergoing LT.

Disclosures:Norah Terrault - Advisory Committees or Review Panels: Eisai, Biotest; Con-sulting: BMS, Merck; Grant/Research Support: Eisai, Biotest, Vertex, Gilead, AbbVie, Novartis, MerckSanjaya K. Satapathy - Advisory Committees or Review Panels: GileadElizabeth C. Verna - Advisory Committees or Review Panels: Gilead; Grant/Research Support: Salix, MerckThomas D. Schiano - Advisory Committees or Review Panels: vertex, salix, merck, gilead, pfizer; Grant/Research Support: massbiologics, itherxSher Linda - Grant/Research Support: BiotestJohn M. Vierling - Advisory Committees or Review Panels: Abbvie, Bristol-Mey-ers-Squibb, Gilead, Hyperion, Intercept, Janssen, Novartis, Merck, Sundise, HepQuant, Salix; Grant/Research Support: Abbvie, Bristol-Meyers-Squibb, Eisai, Gilead, Hyperion, Intercept, Janssen, Novartis, Merck, Sundise, Ocera, Mochida; Speaking and Teaching: GALA, Chronic Liver Disease Foundation, ViralEdJacqueline G. O’Leary - Consulting: Gilead, JansenLaura M. Kulik - Advisory Committees or Review Panels: Bayer/ Onyx; Grant/Research Support: Bayer/Onyx; Speaking and Teaching: Bayer/Onyx, Nordion, GileadShailesh Chavan - Employment: Biotest PharmaceuticalsChristopher J. Dougherty - Employment: Biotest Pharmaceuitcals CorporationThe following people have nothing to disclose: George Therapondos, Roshan Shrestha, Jeffrey Campsen, James Spivey, Jens Rosenau, Kalyan R. Bhamidi-marri, Lewis W. Teperman, Gerond Lake-Bakaar, Fredric D. Gordon, Daniel Maluf

21High rates of virological response and major clinical improvement during sofosbuvir and daclatasvir-based regimens for the treatment of fibrosing cholestatic HCV-recurrence after liver transplantation: The ANRS CO23 CUPILT studyVincent Leroy1, Jérôme Dumortier2, Audrey Coilly3, Mylene Sebagh3, Claire Fougerou-Leurent4, Sylvie Radenne14, Danielle Botta5, Francois Durand6, Christine Silvain7, Pascal Lebray8, Pau-line Houssel-Debry4, Nassim Kamar9, Louis d’Alteroche10, Yvon Calmus11, Inga Bertucci12, Georges-Philippe Pageaux13, Jean-Charles Duclos-Vallee3; 1CHU de Grenoble, Grenoble, France; 2Hôpital Edourat Herriot, Lyon, France; 3Hôpital Paul Brousse, Paris, France; 4CHU de Rennes, Rennes, France; 5APHM, Mar-seille, France; 6Hôpital Beaujon, Clichy, France; 7CHU de Poitiers, Poitiers, France; 8Hôpital LPS, Paris, France; 9CHU de Toulouse, Toulouse, France; 10CHU de Tours, Tours, France; 11Hôpital St-An-toine, Paris, France; 12ANRS, Paris, France; 13CHU de Montpel-lier, Montpellier, France; 14Hôpital de la croix rousse, Lyon, FranceFibrosing cholestatic hepatitis (FCH) is a rare but severe form of HCV-recurrence following liver transplantation (LT) leading to poor short term survival. Therapeutic options are limited.

Study aims were to assess efficacy and tolerance of sofosbuvir (SOF) and daclatasvir (DCV)-based regimens in this setting. Methods: The CUPILT study is a prospective nationwide cohort including patients with HCV-recurrence following LT treated by new antivirals. The present work focused on 21 patients diagnosed with FCH and included between Oct 2013 and Feb 2014. FCH diagnosis was based on strict criteria includ-ing histological review by an expert pathologist. Treatment regimens were prescribed at investigator’s discretion. Patients were followed at W0, W1, W2, W3, W4, W6, W8 and W12. Results: FCH was diagnosed after a median duration of 6 months [range 1-18] following LT and therapy started at 10 months [2-38] post LT. Features of patients at W0 were: median age: 53 years [36-67], men: 81%, G1: 76%, G3: 10%, G4: 14%, ALT: 128 IU/L [46-588], gGT: 595 IU [86-5,511], bilirubin : 6.0 mg/dL [0.6-19], albumin : 3.2 g/dL [1.6-4.0], HCV RNA : 6.9 log IU/ml [4.6-8.4]. Ascites was observed in 8 (38%) patients. Four (19%) were HIV-co-infected and 14 (67%) failed to antiviral therapy containing protease inhibitors in 9 (43%) cases. The following regimens were used: Peg-IFNa + SOF + RBV (n=2), SOF + RBV (n=6), SOF + DCV (n=1) and SOF + DCV + RBV (n=12) for 24 weeks. All patients were alive without re-transplantation at W12. HCV RNA was not detectable at W2 and W4 in 1 (5%) and 3 (14%) patients, respectively. At W12, 20 (95%) patients had HCV RNA <15 IU/ml and 17 (81%) were not detectable. Early viral kinetics was not influenced by treatment regimens. The rates of ALT and gGT normalization at W12 were 76% and 52%, respectively. Median bilirubin serum levels rapidly decreased from 6.0 to 3.6 at W1 and 2.6 mg/dL at W2. The median time to achieve bilirubin levels below 2 mg/dL was 6 weeks. At week 12, 19 (90%) of patients had bilirubin below 2 mg/dl, and 13 (69%) below 1 mg/dL. Albumin levels increased from 31 to 36 g/L at W12. This was accompanied by clinical improvement including nutritional status (weight gain from 67.5 to 70.0 Kg). Ascites disappeared in 4/8 patients, but remained stable in 2 patients who had initially refractory ascites. Complete clinical response (no ascites and bilirubin < 2 mg/dL) was observed at W12 in 15 (71%) patients. Tolerance was satisfactory. Con-clusion: SOF and DCV based regimens show promising results combining high rates of virological response and major clinical improvement at W12. Durability of virological and clinical response will be presented.Disclosures:Vincent Leroy - Board Membership: roche, merck, gilead, bms, roche, merck, gilead, bms, roche, merck, gilead, bms, roche, merck, gilead, bms; Consulting: jansen, jansen, jansen, jansen; Grant/Research Support: roche, gilead, bms, roche, gilead, bms, roche, gilead, bms, roche, gilead, bms; Speaking and Teach-ing: bms, merck, gilead, roche, bms, merck, gilead, roche, bms, merck, gilead, roche, bms, merck, gilead, rocheJérôme Dumortier - Board Membership: Novartis, Astellas, Roche; Consulting: Novartis; Grant/Research Support: Novartis, Astellas, Roche, MSD, GSKAudrey Coilly - Speaking and Teaching: Gilead, BMS, Janssen, MSD, Roche, Novartis, AstellasFrancois Durand - Advisory Committees or Review Panels: Astellas, Novartis; Speaking and Teaching: GileadPascal Lebray - Grant/Research Support: Merck, astellas; Speaking and Teach-ing: Janssen, MSD, GileadGeorges-Philippe Pageaux - Advisory Committees or Review Panels: Roche, Roche, Roche, Roche; Board Membership: Astellas, Astellas, Astellas, AstellasThe following people have nothing to disclose: Mylene Sebagh, Claire Foug-erou-Leurent, Sylvie Radenne, Danielle Botta, Christine Silvain, Pauline Hous-sel-Debry, Nassim Kamar, Louis d’Alteroche, Yvon Calmus, Inga Bertucci, Jean-Charles Duclos-Vallee

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22Striking differences in wait-listing trends between patients with viral hepatitis B and C: implications of effective anti-viral therapyJennifer A. Flemming1, W. Ray Kim2, Carol Brosgart3, Norah Ter-rault3; 1Medicine and Public Health Services, Queen’s University, Kingston, ON, Canada; 2Medicine, Stanford University, Palo Alto, CA; 3Medicine, University of California San Francisco, San Fran-cisco, CATrends in wait-listing (WL) for liver transplantation (LT) reflect the changing epidemiology of the cirrhotic population. We aimed to analyze trends in LT WL for viral hepatitis in the United States (US). Methods: Using the scientific registry of transplant recipi-ents database from 2003-2013, we identified adults WL for LT due to hepatitis C (HCV) and hepatitis B (HBV), with non-alco-holic steatohepatitis (NASH) as a comparator. The indication for WL was defined either as end-stage liver disease (ESLD) if the MELD at WL was ≥ 15 or hepatocellular carcinoma (HCC). Standardized annual incidence rates of WL based on etiol-ogy and indication were calculated and time trends analyzed using Poisson regression to calculate incidence rate ratios (IRR). Results: 42,855 individuals were identified (HCV 74%, NASH 18%, HBV 8%), 71% male, median age 55 yrs (IQR 51 – 61). The age and sex adjusted incidence of LT WL increased annually for HCV (IRR 1.03, 95% CI 1.02-1.03, P <.001) and NASH (IRR 1.11, 95% CI 1.10-1.12, P <.001) and decreased for HBV (IRR 0.987, 95% CI 0.976-0.999, P = 0.027). WL for ESLD increased by 11% per year in NASH (IRR 1.11, 95% CI 1.10-1.12, P <.001) while it decreased by 4% per year in HBV and 1% in HCV (HBV IRR 0.96, 95% CI 0.94-0.97, P <.001; HCV IRR 0.99, 95% CI 0.98-0.99, P <.001; figure). WL for HCC increased by 10% per year for HCV (IRR 1.10, 95% CI 1.09-1.11, P <.001), and 15% for NASH (IRR 1.15, 95% CI 1.12-1.17, P <.001), but was stable in HBV (HBV IRR 1.01, 95% CI 0.99-1.03, P = 0.261, figure). Conclusions: While these trends in WL for HBV likely reflect the success of anti-viral therapy, the effect of anti-HCV therapy is yet to become appar-ent, especially for HCC. Future reduction in the need for WL for HCV may help accommodate the growing need for LT for NASH in the US.

Disclosures:W. Ray Kim - Consulting: Bristol Myers Squibb, Gilead SciencesCarol Brosgart - Board Membership: Tobira Therapeutics; Consulting: Dynavax; Stock Shareholder: Alios BiopharmaNorah Terrault - Advisory Committees or Review Panels: Eisai, Biotest; Con-sulting: BMS, Merck; Grant/Research Support: Eisai, Biotest, Vertex, Gilead, AbbVie, Novartis, MerckThe following people have nothing to disclose: Jennifer A. Flemming

23Sofosbuvir/Daclatasvir Therapy for Recurrent Hepatitis C after Liver Transplantation: Preliminary report from the parisian centersFilomena Conti1, Pascal Lebray2, Astris Schielke1, Helene Reg-nault2, Dominique Thabut2, Daniel Eyraud2, Armelle Poujol-Rob-ert1, Olivier Chazouillères1, Yvon Calmus1; 1Liver Transplant unit, Hôpital Saint Antoine, APHP, Paris, France; 2Hepatogastroenterol-ogy and Liver Transplant units, Paris VI-Pitié-Salpêtrière Hospital, Paris, FranceBackground: Recurrence of hepatitis C (HCV) infection is the most common cause of graft loss and mortality in HCV-infected liver-transplant recipients. Interferon-based antiviral regimens, including those utilizing protease inhibitors, are poorly toler-ated after transplantation and achieve lower sustained viro-logic response rates (SVR) than in nontransplant patients. The nucleotide HCV polymerase inhibitor sofosbuvir (SOF) and the HCV nonstructural protein NS5A inhibitor daclatasvir (DTV) have a high barrier to resistance, lack interactions with immu-nosuppressive agents, and have a favourable safety profile. Methods: In this ongoing single-arm, open-label interferon-free pilot study, patients with recurrent HCV infection (any HCV genotype) after liver transplantation received up to 24 weeks of SOF 400 mg and DTV 60 mg/day. Patients were ≥18 years of age and at least 6 months post-transplantation. The primary efficacy endpoint of this preliminary report was viro-logic response (VR) (HCV RNA <15 IU/mL) 2, 4 and 8 weeks after initiation of study treatment. Results: 55 patients (16 GT 1a, 18 GT 1b, 1 GT1g, 6 GT 2, 8 GT 3, 6 GT 4) have been enrolled. Mean age was 56 ± 8.7 years, 85 % were males, 91 % have been previously treated. The mean baseline viral load was 6.35 ± 2.95 log10 IU/mL [range 2.08- 7.50 log10 IU/mL], 18 patients were F3/F4 and 4 had fibrosing cholestatic hepatitis. By week 2, 4 and 8 of treatment, VR was 17, 50 and 85 %, respectively, while the viral load decreased to 3.14 ± 2.95, 0.90 ± 0.80 and 0.23 ± 0.25, respectively. Prothrombin rate, albumin and creatinin levels remained unchanged during the 8 first weeks of treatment.There was no episode of acute or chronic rejection. No dose adjustment of immunosuppression was required. The most common adverse events were fatigue, arthralgia, headache, and diarrhea. 2 patients experienced renal failure possibly related to antiviral therapy. No patient discontinued treatment due to adverse event. Conclusions: SOF/DTV therapy was well tolerated and achieved an early VR of 50 and 85 % at W4 and W8 in these difficult to cure patients. Sustained VR will be presented at the Liver Meeting.Disclosures:Pascal Lebray - Grant/Research Support: Merck, astellas; Speaking and Teach-ing: Janssen, MSD, GileadOlivier Chazouillères - Consulting: APTALIS, MAYOLY-SPINDLERThe following people have nothing to disclose: Filomena Conti, Astris Schielke, Helene Regnault, Dominique Thabut, Daniel Eyraud, Armelle Poujol-Robert, Yvon Calmus

24Therapeutic misadventure with acetaminophen: pro-spective evaluation of prerequisites, characteristics and outcomeAlexandre Louvet1,2, Charlotte Vanveuren1, Sébastien Dha-rancy1,2, Amélie Cannesson1,2, Florent Artru1,2, Guillaume Las-sailly1,2, Valerie Canva-Delcambre1, Philippe Mathurin1,2; 1Service des maladies de l’appareil digestif, Hôpital Huriez, Lille, France; 2Unité 995, INSERM, Lille, FrancePrerequisites leading to development of the so-called “therapeu-tic misadventure with acetaminophen” have never been eval-

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uated in a prospective study with sufficient sample size. Such study would enable to characterize not only outcome but also pattern of acetaminophen intake. Methods: All patients admit-ted with severe acetaminophen-related hepatitis were included prospectively. Patients were divided into acute hepatitis related to acetaminophen overdose (AO) or to acetaminophen use at therapeutic dose (≤6g/day), termed acetaminophen thera-peutic misadventure (ATM). We defined chronic drinkers by a daily alcohol consumption of ≥30g/day. Results: From 2002 to 2014, 271 patients were included, 205 with AO and 66 with ATM. Only patients who were chronic alcohol drinkers (89.4%) or who had starved for several days (9.6%) developed ATM. As expected, acetaminophen intake was 16 g in AO vs. 3.15g in ATM (p<0.0001). In 70% of patients, acetamino-phen intake was below the daily 4g-dose classically regarded as safe. In contrast to AO, no patient developed ATM after a single intake of acetaminophen as confirmed by a median time of intake at 4 days in ATM vs. 1 day in AO (p<0.0001). At admission, patients with ATM were older than AO (age 44 vs. 30.1 years; p<0.0001) and had a deeper impairment of liver function (prothrombin time 30.7 vs. 22.1s, p=0.0003; albumin 32.7 vs. 38.1 g/l, p<0.0001; bilirubin 47.1 vs. 25 mg/l, p<0.0001; lactate 3.31 vs. 1.95 mmol/l, p=0.009; creatinine 9.8 vs. 8.7 mg/l, p=0.097). Transaminases were as elevated in ATM as in AO: AST 4138 vs. 3983 IU/l (p=0.1), ALT 2416 vs. 3831 IU/l (p=0.22). In ATM and AO, males were 36.4% and 43.4% (p=0.31) and percentage of chronic drinkers was 89.4 vs. 36.6% (p<0.0001), respectively. After 3 days, a drastic drop by 67% vs. 50.2% (p<0001) in ALT and by 93.6 vs. 89.4% (p=0.02) in AST was observed in ATM and AO respectively. One-month survival was 84.6% in ATM and 92.6% in AO (p=0.05). On overall patients, among factors associated with survival in univariate analysis, only the number of King’s college criteria (risk ratio 3.15, 95%CI: 2.04-4.88, p<0.0001) independently predicted 1-month mortality, whereas albumin, age and alcohol consumption did not (p= 0.13, 0.41 and 0.75 respectively). Conclusion: Therapeu-tic misadventure occurs after several days of acetaminophen intake at a dose <6g/day and is mainly observed in chronic alcohol consumers and rarely after prolonged starvation. This entity has a worse outcome as compared to acetaminophen overdose. Clinicians must be warned that repeated prescription of acetaminophen must be carefully monitored in patients with risk factors of misadventure.Disclosures:Sébastien Dharancy - Board Membership: NOVARTIS; Speaking and Teaching: ROCHE, ASTELLASPhilippe Mathurin - Board Membership: Schering-Plough, Janssen-Cilag, BMS, Gilead, Abvie; Consulting: Roche, Bayer, BoehringerThe following people have nothing to disclose: Alexandre Louvet, Charlotte Vanveuren, Amélie Cannesson, Florent Artru, Guillaume Lassailly, Valerie Can-va-Delcambre

25Hepatocyte-derived extracellular vesicles released during lipotoxicity regulate hepatic stellate cell pheno-type via delivery of specific miRNAs and modulation of PPAR-γ expressionDavide Povero1, Nadia Panera2, Akiko Eguchi1, Anna Alisi2, Vale-rio Nobili2, Ariel E. Feldstein1; 1Department of Pediatrics, Uni-versity of California San Diego, La Jolla, CA; 2Hepato-Metabolic Disease Unit and Liver Research Unit, Bambino-Gesu’ Children’s Hospital, IRCCS, Roma, ItalyBackground. Hepatic stellate cell (HSC) activation plays a pivotal role in liver fibrosis and disease progression in non-alcoholic fatty liver disease (NAFLD). The triggers for HSC acti-

vation in this condition remain poorly understood. We have recently demonstrated that during lipotoxicity, hepatocytes release extracellular vesicles (EVs) enriched in miRNAs (Sci-ence Sig. Oct 2013). Our aim is to investigate if extracellular vesicles released by hepatocytes during lipotoxicity may modu-late hepatic HSC phenotype by delivering specific microRNAs. Methods. Human hepatoma cells (HepG2), and primary mouse hepatocytes were exposed to the saturated free fatty acid (FFA) palmitic acid for up to 24 hrs. EVs and EV-free supernatant were isolated from cell-free supernatants by ultracentrifuga-tion and quantitated by flow cytometry. HSC chemotaxis and chemokinesis were assessed by Boyden’s chamber and wound healing assay, respectively. HSC proliferation was assessed by BrDu-FITC staining and quantitation of pro-fibrogenic tran-scripts was performed for cell activation. EVs internalization and delivery of miRNAs into HSC was addressed by immuno-fluorescence. Specific PPAR-γ-targeting miRNAs identified and quantified in EVs and HSCs by qPCR. Depletion of miRNAs from EVs was achieved by anti-miRNA and specific siRNA on maternal cells. A functional analysis of miRNA was assessed by miRNA mimics. Results. Hepatocyte-derived EVs released during lipotoxicity are efficiently internalized by HSCs result-ing in their activation, as shown by marked up-regulation of pro-fibrogenic genes, such as Collagen-I, α-SMA and TIMP-2, proliferation (EVs vs. EVs-free supernatant, p<0.04), chemo-taxis (EVs vs. EV-free supernatant, p<0.001) and chemokinesis (EVs vs. EVs-free supernatant, p<0.002), mainly after 16-24 hrs. These changes were associated with suppression of PPAR-γ expression in HSC. EVs internalization results in delivery of their miRNA content into HSCs. Lipotoxic hepatocyte-derived EVs miRNA content included various miRNAs that are known inhibitors of PPAR-γ expression with miR-128a being the most effective. Further loss- and gain-of-function studies identified miR-128a as a central modulator of the effects of EVs on PPAR-γ inhibition and HSC activation. Conclusion. Our study demon-strates that EVs released by hepatocytes during lipotoxicity are critical signals that contribute to HSC activation in a process involving delivery of specific miRNAs and modulation of PPAR-γ expression. These results uncover a novel miRNA-regulated pathway committing HSC activation during lipotoxicity and have important implications for development of therapeutic strategies for patients with NAFLD.Disclosures:Akiko Eguchi - Grant/Research Support: Gilead The following people have nothing to disclose: Davide Povero, Nadia Panera, Anna Alisi, Valerio Nobili, Ariel E. Feldstein

26Sphingosine kinase containing exosomes regulate hepatic stellate cell activationRuisi Wang, Sheng Cao, Usman Yaqoob, Thiago de Assuncao, Vijay Shah; Mayo Clinic, Rochester, MNBackground/Aims: Hepatic stellate cell (HSC) activation is required for fibrogenesis therefore understanding mechanisms governing HSC activation are important. Exosomes are cell derived extracellular vesicles thought to promote intercellular communication by interacting with recipient cells to deliver specific content. The aim of this study was to determine whether exosomes contribute to HSC activation during liver fibrosis. Methods/Results: Exosomes were isolated from human serum and conditional media of TSEC (immortalized mouse liver endothelial cells) by differential ultracentrifugation, and char-acterized based on size (90-110 nm) and marker (TSG101, LAMP1, CD63 and CD81) characteristics. Incubation of LX2 HSC cell line with human serum derived exosomes was asso-