studies

222A AASLD ABSTRACTS HEPATOLOGY, October, 2015
advantage of our HCV culture system in primary human adult
hepatocytes (PHH), which, contrary to the widely used hepatocarcinoma-derived
Huh-7 sublines, retain the liver metabolism
of ethanol and secrete authentic VLDL and LVP. METHODS:
PHH were infected with the HCV strain JFH1 or a chimeric
virus derived thereof and treated with increasing doses of ethanol
(0-100 mM) for 2 weeks. Cultures were monitored for
HCV genome replication (negative strand RT-qPCR), viral load
(clinically used test), production of infectious virus (titration by
focus-formation assay), and VLDL secretion (apolipoprotein B
ELISA). The density of viral particles was assessed by isopycnic
iodixanol ultracentrifugation. RESULTS: Ethanol exposure of
HCV-infected PHH caused a time- and dose-dependent increase
in the viral load, comparable to that reported in clinical studies.
Most strikingly, it caused an even greater increase in the
infectious titer but did not significantly affect the viral genome
replication, thus pointing to an effect on virus morphogenesis.
This effect was not seen in Huh-7.5.1 cells treated in parallel,
suggesting that it involves the liver metabolism of ethanol. The
higher specific infectivity of HCV particles produced by PHH
in the presence of ethanol correlated with lower mean buoyant
density, consistent with triglyceride enrichment. Finally, in
either HCV-infected or naïve PHH, addition of ethanol caused
a time-dependent increase in VLDL secretion. CONCLUSION:
This study in the relevant PHH model reveals that ethanol via its
metabolites increases the production of HCV particles of lowest
buoyant density and highest infectivity, i.e., LVP, most likely
due to the impact of ethanol on triglyceride metabolism that
results in increased VLDL secretion. Drugs targeting this host
metabolic pathway may be useful in difficult-to-treat alcoholic
patients, often poorly compliant with therapy and therefore at
risk for resistance if treated by direct antivirals only. VP & CH:
equal contribution.
Disclosures:
The following authors have nothing to disclose: Veronique Pene, Céline Hernandez,
Etienne Blanc, Lynda Aoudjehane, Béatrice Le Grand, Arnaud Carpentier,
Jean-François Méritet, Filomena Conti, Yvon Calmus, Hélène Rouach, Philippe
Podevin, Arielle R. Rosenberg
followed by 15 weeks of combined therapy with 250mg REP
2139-Ca and 180ug Pegasys® and then 33 weeks with
Pegasys® monotherapy. Viremia (HDV RNA and HBV DNA),
HBsAg and anti-HBs are followed every two weeks (Robogene
RT-PCR, Abbott RealTime HBV, Abbott Architect) performed at
the Institute of Virology, University of Duisburg-Essen (Essen,
Germany). HDV RNA is validated on separate test platforms
at the National Genetics Institute (Los Angeles, USA) and the
Institute of Virology, Technische Universität München (Munich,
Germany). Results: REP 2139-Ca treatment is well tolerated
with mild and quickly resolving IV infusion reactions. Serum
HBsAg is currently reduced 1-6 log in 11/12 patients (5 with
serum HBsAg < 1 IU / ml) and HDV RNA is currently reduced
1.5-7 log in 12 /12 patients (undetectable in 6 patients). Anti-
HBs is detected in 10/12 patients, with 6 patients < 10mIU /
ml and after combined exposure to Pegasys®, 5 patients have
substantially increased anti-HBs titers from 50 to > 800 mIU /
ml. In all patients with pre-treatment HBV DNA < 10 IU / ml,
de-repression of serum HBV DNA is observed. Conclusions:
Updated interim data from the REP 301 protocol assessing the
safety and antiviral efficacy of REP 2139 (first in monotherapy
and then with add on Pegasys® at week 16) in 12 Caucasian
patients with chronic HBV / HDV co-infection demonstrated
substantial reductions in serum HBsAg and HDV RNA as well
as appearance of anti-HBs. HDV RNA reductions appear stronger
than HBsAg reductions, suggesting an additional antiviral
mechanism other than the inhibition of subviral particle assembly
may affect HDV directly. REP 2139-Ca may become an
important new therapeutic option for patients with chronic HBV
/ HDV infection.
Disclosures:
Michel Bazinet - Board Membership: REPLICor Inc.; Employment: REPLICor Inc.;
Management Position: REPLICor Inc.; Patent Held/Filed: REPLICor Inc.; Stock
Shareholder: REPLICor Inc.
Andrew Vaillant - Employment: REPLICor; Stock Shareholder: REPLICor
The following authors have nothing to disclose: Victor Pantea, Valentin
Cebotarescu, Lilia Cojuhari, Paulina Jimbei, Jeffrey Albrecht, Peter Schmid, Hadi
Karimzadeh, Michael Roggendorf
31
Update on the safety and efficacy of REP 2139 monotherapy
and subsequent combination therapy with
pegylated interferon alpha-2a in chronic HBV / HDV
co-infection in Caucasian patients
Michel Bazinet 1 , Victor Pantea 2 , Valentin Cebotarescu 2 , Lilia
Cojuhari 3 , Paulina Jimbei 3 , Jeffrey Albrecht 4 , Peter Schmid 4 , Hadi
Karimzadeh 5 , Michael Roggendorf 5 , Andrew Vaillant 1 ; 1 Replicor
Inc., Montreal, QC, Canada; 2 N. Testemitanu State University of
Medicine and Pharmacy, Chisinau, Moldova (the Republic of);
3 Toma Ciorba Infectious Clinical Hospital, Chisinau, Moldova
(the Republic of); 4 National Genetics Institute, Los Angeles, CA;
5 Institure of Virololgy, Technische Universität München, Munich,
Germany
Introduction: HBV / HDV co-infection causes rapid progression
of liver disease and with no approved therapy, presents a
significant unmet medical need. Nucleic acid polymers (NAPs)
block HBV subviral particle assembly and release from infected
hepatocytes and can eliminate serum HBsAg. As the NAP REP
2139 was previously been shown to clear serum HBsAg and
improve the ability of immunotherapy to elicit SVR in Asian
patients with HBV, its activity in combination with Pegasys® in
HBV / HDV co-infected Caucasian patients is currently being
examined. Methods: In a phase II proof of concept trial (REP
301; NCT02233075), patients with chronic HBV / HDV co-infection
received once weekly dosing of 500mg REP 2139-Ca
(calcium chelate complex) by 2h IV infusion for 15 weeks,
32
Reductions in cccDNA under NUC and ARC-520 therapy
in chimpanzees with chronic hepatitis B virus infection
implicate integrated DNA in maintaining circulating
HBsAg
Christine I. Wooddell 1 , Deborah Chavez 2 , Jason E. Goetzmann
3 , Bernadette Guerra 2 , Ryan M. Peterson 1 , Helen Lee 2 ,
Julia O. Hegge 1 , Robert Gish 4 , Stephen Locarnini 5 , Christopher
R. Anzalone 1 , Robert E. Lanford 2 , David L. Lewis 1 ; 1 Arrowhead
Madison, Arrowhead Research Corporation, Madison, WI; 2 Texas
Biomedical Research Institute, San Antonio, TX; 3 New Iberia
Research Center, University of Louisiana at Lafayette, New Iberia,
LA; 4 Department of Medicine, Stanford University Medical Center,
San Diego, CA; 5 Victorian Infectious Diseases Reference Laboratory,
Melbourne, VIC, Australia
Background: RNAi therapeutic ARC-520 designed to target
all cccDNA-derived transcripts reduces viral antigenemia for
>1 month after single doses in HBV patients. Here we report
the effect of multiple ARC-520 doses on hepatic HBV DNA
and RNA in HBV chimps. Methods: 9 chimps (5 M, 4 F; 9-37
yrs) received 6-11 monthly injections of ARC-520 concurrent
with NUC therapy. 5 were HBeAg-positive (HBeAg+), baseline
DNA 8-9 log 10
IU/mL serum, and 4 were HBeAg-negative
(HBeAg-), ≤3 log 10
IU/mL. Chimps received NUCs for
8-24 weeks prior to ARC-520 dosing. Liver biopsies from 8
chimps were taken at baseline, completion of NUC lead-in
and on study. HBV DNA, +/- plasmid-safe DNase digestion to