A Multicentre, Phase II, Open-Label, Randomised Controlled Trial of Repeated Autologous Infusions of Granulocyte Colony Stimulating Factor Mobilised Cd133 + Bone Marrow Stem Cells in Patients with Cirrhosis

ORAL PRESENTATIONS

JOURNAL OF HEPATOLOGY

Saturday, 16 April 2016

General session 3 and awards 2 GS13 LEDIPASVIR/SOFOSBUVIR FOR 12 OR 24 WEEKS IS SAFE AND EFFECTIVE IN KIDNEY TRANSPLANT RECIPIENTS WITH CHRONIC GENOTYPE 1 OR 4 HCV INFECTION M. Colombo1, A. Aghemo1, L. Liu2, R. Hyland2, C. Yun2, D. Brainard2, J. McHutchison2, M. Bourlière3, M. Peck-Radosavljevic4, M. Manns5, S. Pol6. 1Fondazione IRCCS Ca’ Granda Ospedale Maggiore Policlinico, Milan, Italy; 2Gilead Sciences Inc., Foster City, United States; 3Hôpital Saint Joseph, Marseilles, France; 4Medical University of Vienna, Vienna, Austria; 5Hannover Medical School, Hannover, Germany; 6Hôpital Cochin, Paris, France E-mail: [email protected] Background and Aims: Interferon (IFN) and ribavirin (RBV) for the treatment of chronic hepatitis C (HCV) in kidney transplant recipients is complicated by the risk of the allograft rejection and poor tolerability. We evaluated the safety and efficacy of the IFN-free, RBV-free regimen of ledipasvir/sofosbuvir (LDV/SOF) in chronic genotype (GT) 1 or 4 HCV infected kidney transplant recipients. Methods: Kidney transplant recipients with chronic GT1 or GT4 HCV infection, treatment-naïve and treatment-experienced, with or without compensated cirrhosis were randomized 1:1 at 5 sites in Europe to receive LDV/SOF (90 mg/400 mg) for 12 or 24 weeks. Randomization was stratified by HCV genotype, treatment history and presence or absence of cirrhosis. Cirrhosis was determined by liver biopsy (Metavir score = 4 or Ishak score ≥5), Fibroscan® >12.5 kPa, or Fibrotest® >0.75 and APRI >2. A pretreatment creatinine clearance <40 mL/min was an exclusionary criterion. The primary endpoint was SVR12. Results: 114 patients were randomized and treated; median age was 53, 58% were male, 94% were white, 72% carried the non-CC IL28B allele, 91% had genotype 1 infection, 69% were treatment-naïve, and 15% had compensated cirrhosis. The median eGFR was 56 mL/min (range 35–135 mL/min). All 92 patients with SVR4 data available achieved SVR4 including a patient discontinuing treatment at Week 4 due to an AE. SAEs were reported in 12 (11%) patients; 3 were assessed as treatment related: syncope, pulmonary embolism, and blood creatinine increased. The most frequent AEs were headache (19%), asthenia (13%), and fatigue (10%). Conclusions: Administration of LDV/SOF for 12 or 24 weeks in patients with chronic HCV genotype 1 or 4 patients who have undergone kidney transplant was safe and highly effective with an SVR4 rate of 100%. Treatment was well-tolerated. SVR12 data for all patients will be presented. GS14 TM6SF2 AND NCAN WILDTYPE TOGETHER WITH SEVERAL LONGCHAIN PHOSPHATIDYLCHOLINES ARE LINKED TO A HEALTHY LIVER IN OBESE SUBJECTS A. Feldman1, S.K. Eder1, T.K. Felder2, L. Kedenko1, B. Paulweber1, A. Stadlmayr3, C. Datz3, E. Aigner1. 1First Department of Medicine; 2 Department of Laboratory Medicine, Paracelsus Medical University, Salzburg; 3Department of Internal Medicine, General Hospital, Oberndorf, Austria E-mail: [email protected]

Background and Aims: A small proportion of obese individuals do not have evidence of metabolic complications. We aimed to provide a clinical, metabolic and genetic description of obese subjects without evidence of liver disease. Methods: Among 3000 Caucasian subjects undergoing a health screening program 173 participants were identified and allocated to one of 3 groups according to BMI, metabolic syndrome (MetS), biochemical liver tests and hepatic steatosis on ultrasound: lean control subjects (BMI ≤ 25 kg/m², no steatosis, normal liver tests, no MetS, n = 71), obese healthy (BMI ≥ 30 kg/m², no steatosis, normal liver tests, no MetS, n = 51), obese NAFLD (BMI ≥ 30 kg/m², steatosis, MetS, n = 61). All subjects underwent a detailed clinical, genetic and metabolic evaluation, including serum metabolomics. Results: In our study, obese healthy subjects were more likely to be female compared to the other groups (p < 0.001) and were not different from lean healthy subjects with regard to glucose parameters such as OGTT, fasting glucose and HBA1c. Obese healthy subjects had a significantly lower rate of the mutant TM6SF2 variant compared to obese NAFLD (2/51 vs. 12/61; p = 0.019) and lean controls (2/51 vs. 13/ 71, p = 0.023). The NCAN mutant variant was less frequent in obese healthy subjects compared to obese NAFLD subjects (2/51 vs. 12/61: p = 0.019). No differences were found in the carrier rates of PNPLA3 between groups. Obese healthy subjects had higher leptin and adiponectin compared to lean healthy (p = 0.02; p < 0.001) and obese NAFLD subjects (p < 0.001; p = 0.03) and lower TNFα and IL-6 concentrations compared to obese NAFLD subjects (p < 0.001; p = 0.01). The metabolomics study identified various phosphatidylcholines (PC ae C44:6, PC ae C42:5, PC aa C40:4; FDR corrected p-value < 0.001) as well as lysine, glycine and isoleucine (FDR < 0.001) to be associated with the healthy liver phenotype in obese subjects. Conclusions: Obese subjects with evidence of a healthy liver are characterized by intact glucose homeostasis, lower concentrations of proinflammatory cytokines and higher adiponectin and leptin concentrations compared to obese subjects with NAFLD. Wildtype TM6SF2 and NCAN alleles along with long chain phosphatidylcholines are protective against the development of fatty liver in obesity. GS15 A MULTICENTRE, PHASE II, OPEN-LABEL, RANDOMISED CONTROLLED TRIAL OF REPEATED AUTOLOGOUS INFUSIONS OF GRANULOCYTE COLONY STIMULATING FACTOR MOBILISED CD133 + BONE MARROW STEM CELLS IN PATIENTS WITH CIRRHOSIS P.N. Newsome1, R. Fox2, A. King2, N. Than2, J. Moore3, C. Corbett2, S. Townsend4, D. Barton4, H. Beard2, A. Atkinson5, C. Bienek5, N. McGowan5, J. Campbell5, J. Keeley4, J. Thomas6, K. Guo4, D. Hull4, I. Guha7, D. Hollyman8, D. Stocken9, C. Yap4, S. Forbes6. 1National Institute for Health Research (NIHR) Birmingham Liver Biomedical Research Unit and Centre for Liver Research; 2University of Birmingham, Birmingham; 3University of Edinburgh, Edinburgh; 4NIHR Birmingham Liver Biomedical Research Unit Clinical Trial group (EDD), CRUK Clinical Trials Unit, University of Birmingham, Birmingham; 5SNBTS; 6MRC Scottish Centre for Regenerative Medicine, University of Edinburgh, Edinburgh; 7National Institute for Health Research Biomedical Research Unit in Gastrointestinal and Liver Diseases, University of Nottingham, Nottingham; 8Cellular and Molecular Therapies, NHS Blood and Transplant, Birmingham; 9Newcastle Clinical Trial Unit, University of Newcastle, Newcastle, United Kingdom E-mail: [email protected]

Journal of Hepatology 2016 vol. 64 | S183–S212 © 2016 All rights reserved.

ORAL PRESENTATIONS Background and Aims: Liver disease mortality and morbidity are rising and liver transplantation is limited by organ availability. Small scale human studies showed that autologous stem cell therapy whilst safe and readily available needed a well conducted randomised controlled trial. Methods: Patients with liver cirrhosis and MELD score 11.5–15.5 were randomised to Arm 1 – Conservative management; Arm 2 – Treatment with granulocyte colony stimulating factor (G-CSF) (Lenograstim) 15 mcg/kg od for 5 days or Arm 3 – Treatment with G-CSF 15 mcg/kg od for 5 days followed by leukapheresis, CD133+ cell isolation and infusion of freshly isolated CD133+ cells immediately and frozen doses at days 30/60 via peripheral vein (0.2 × 106 cells/kg). Co-Primary objectives were to demonstrate improvement in the severity of liver disease (change in MELD) at 3 months, and its trend over time, across Arms. To detect a 0.8 point reduction in MELD with a 2-sided α = 5% (α = 0.1 split equally) and 80% power required 27 participants per Arm (81 in total). Analyses were performed on a modified intention-to-treat basis. Results: Mean (SD) age was 55 (8.8), 53 were male, 37 had alcoholrelated liver disease and 10 had HCV. One patient withdrew fully from Arm 1 (day 30) and two from Arm 3 (day 8 and 98). 19 (68%) Arm 3 patients received the intended dose of CD133+ cells. 27 (96%) Arm 3 patients and 24 (92%) Arm 2 patients received the intended dose of GCSF. The primary outcome measures were not met. Median (IQR) change in MELD for the control, GCSF, and GCSF + cell arms was −0.3 (−1.5, 1.1), −0.5 (−1.4, 0.5) & −0.7 (−1.3, 1.0) respectively. There was no evidence of a difference in change in MELD between control and Arm 2 ( p = 0.78) or Arm 3 (p = 1.0). Moreover, there was no evidence of a difference in trend of change in MELD over time. None of the conclusions differed following a per-protocol analysis. No evidence of a difference was found in UKELD or the individual components of liver function (bilirubin, albumin, INR and creatinine). Median (IQR) change in Fibroscan (to day 90) were 0.0 (−1.6, 8.6), 0.0 (−13.0, 12.0), −0.1 (−6.6, 9.8). There was no evidence of a difference in change in Chronic liver disease questionnaire domains from baseline to day 90. Two Arm 1 patients, three Arm 2 and seven Arm 3 had 1 or more serious adverse events in the 90-day period. Conclusions: In this large, powered, randomised controlled trial GCSF and purified haematopoietic stem cells did not have an impact on liver function or fibrosis. GS16 DEVELOPMENT OF A NEW HEPATOPROTECTIVE AND PROREGENERATIVE MOLECULE BASED ON FIBROBLAST GROWTH FACTOR 15/19 G. Álvarez-Sola1, I. Uriarte2,3, M.U. Latasa1,3, M. Elizalde1, R. Urtasun1,3, C.M. Rodríguez-Ortigosa1–3, J. Prieto1, M.G. Fernández-Barrena1–3, F.J. Corrales1–3, P. Berraondo3,4, C. Berasain1–3, M.A. Ávila1–3. 1Division of Hepatology, CIMA. University of Navarra; 2CIBERehd; 3IDISNA; 4 Division of Immunology, CIMA. University of Navarra, Pamplona, Spain E-mail: [email protected] Background and Aims: The liver has great regenerative ability after acute injury or partial resection. However some pathological conditions such as fatty liver can compromise this capacity. Hepatic steatosis is associated with cholestasis derived from lipotoxicity. A potential source of hepatoprotective molecules can be found in the mechanisms that mediate liver regeneration. Fibroblast growth factor 15 (FGF15), FGF19 in humans, (FGF15/19) is an enterokine induced in the ileum by bile salts, being released into the portal circulation. In the liver FGF15/19 represses bile acids synthesis and regulates glucose and lipid metabolism. Previously we showed that FGF15/19 prevents bile salt overload in regenerating liver and behaves as an important proregenerative factor. It is known that cholestasis and steatosis generate endoplasmic reticulum (ER) stress, leading to liver injury and impaired regeneration. To develop a new hepatoprotective molecule based on the fusion of FGF19 and Apolipoprotein A-I (FibApo), a modification that confers hepatotropism and increases protein half-life. S184

Methods: We used parenchymal and biliary epithelial cells treated with palmitic acid (PA) as a lipotoxicity model, mouse models of hepatic steatosis (high fat diet-HFD-, 12 weeks and db/db mice), cholestatic injury model (α-naphthylisothiocyanate-ANIT) and extensive hepatectomy (85%, PH) model. We also used WT and FGF15KO mice. FGF19 and FibApo were administered from adenoassociated viral vectors or as recombinant proteins subcutaneously. Results: FGF19 and FibApo showed cytoprotective effects against lipotoxicity and attenuated unfolded protein response generated by PA-induced ER stress in vitro. In vivo FGF19 and FibApo protected from ANIT induced cholestasis and reduced the intensity of hepatic ER stress. FGF15KO mice fed HFD developed more steatosis and heightened ER stress response than WTs. Ileal FGF15 was induced by tunicamycin, an inductor of ER stress, and FGF15KO mice displayed higher hepatic and ileal ER stress response to the drug. FGF19 and FibApo reduced hepatic steatosis in db/db mice. Db/db mice infected with AAVs expressing FGF19 and FibApo had increased survival after 85% PH compared to control AAV infected mice. In vivo FibApo was more potent than FGF19. Conclusions: FGF19 showed a clear hepatoprotective capacity in acute liver injury models, being able to modulate the intensity of the ER response. This activity is enhanced in the chimeric protein FibApo. GS17 HIGH RATES OF SVR12 IN ADOLESCENTS TREATED WITH THE COMBINATION OF LEDIPASVIR/SOFOSBUVIR K. Schwarz1, K.F. Murray2, P. Rosenthal3, S. Bansal4, C.-H. Lin5, L. Ni6, B. Kanwar6, J. Fraser6, P. German6, D.M. Brainard6, J. Wen7, R. Gonzalez-Peralta8, M.M. Jonas9, W. Balistreri10. 1Johns Hopkins University School of Medicine, Baltimore; 2Seattle Children’s Hospital, Seattle; 3University of California San Francisco, San Francisco, United States; 4Kings College Hospital, London, United Kingdom; 5Children’s Hospital Los Angeles, Los Angeles; 6Gilead Sciences, Inc., Foster City; 7The Children’s Hospital of Philadelphia, Philadelphia; 8University of Florida, Gainesville; 9Children’s Hospital Boston, Boston; 10Cincinnati Children’s Hospital Medical Center, Cincinnati, United States E-mail: [email protected] Introduction: Although direct acting antivirals have transformed HCV treatment of adults, the standard of care for adolescents and younger children is still limited to treatment with pegylated interferon + ribavirin for 48 weeks in those with genotype 1 (GT1) HCV. The aim of this study was to evaluate the safety, efficacy and pharmacokinetics of the fixed dose single tablet regimen, ledipasvir/ sofosbuvir (LDV/SOF), administered for 12 weeks in GT1 HCVinfected adolescent patients. Material and Methods: Treatment-naïve and treatment-experienced adolescent patients aged 12 to less than 18 years old with chronic GT1 HCV were enrolled into this open-label ongoing study to receive 12 weeks of treatment with LDV/SOF 90 mg/400 mg once daily. The primary and secondary efficacy endpoints are SVR12 and SVR4, respectively (HCV RNA < lower limit of quantitation). Safety is assessed by clinical evaluation and laboratory monitoring. Intensive pharmacokinetic (PK) sampling was done on Day 10 in the first 10 patients (PK lead-in) to confirm the appropriateness of the adult dose in the adolescent population. Results: 100 GT1 patients have been enrolled and treated. The majority are GT1a (81%), female (63%), white (90%), treatment-naive (80%), and vertically infected (85%). The mean age is 15 (range 12–17) years. In the PK lead-in, administration of 1 tablet daily of LDV/SOF provided comparable plasma exposures of LDV, SOF, and GS-331007 (SOF primary metabolite) to those observed in adults. 99 patients have completed treatment per protocol with no early discontinuations due to an adverse event; post-treatment follow-up is ongoing. Of the 96 patients who have reached post-treatment week 4, all have achieved SVR4 (100%, 96/96). Similarly, all 78 patients who have reached post-treatment week 12 have achieved SVR12 (100%, 78/78). No serious adverse events (AEs) have been reported. AEs are

Journal of Hepatology 2016 vol. 64 | S183–S212