studies

1014A AASLD ABSTRACTS HEPATOLOGY, October, 2015
tion of A3B deletion. Results: A3B deletions were detected in
30% of Asian patients and 5% of Caucasian patients. We
identified a significant association between A3B deletion and
positive HBeAg status in Asian HBV patients (p = 0.002).
Patients with this deletion were more likely to be HBeAg positive
and remained HBeAg positive at an older age (Figure). A
similar trend was observed among Caucasians patients though
was not significant, likely due to a low A3B allele deletion frequency
(p = 0.2). By analyzing the SNP data from the 1000
Genomes Project, we observed a significant reduction of variation
around A3B deletion region, suggesting a recent positive
selection on A3B deletion allele in Asian, Indian, and Caucasian
populations. Comparing A3B sequences across primates
revealed a long-term positive selection signature, particularly in
the human lineage. Conclusion: Structural variation of the APO-
BEC3B gene is associated with HBeAg status among Asian
patients. The high frequency of A3B deletion allele in Asian
populations may be partially explained by positive selection
forces unrelated to HBV. Further confirmation in independent
patient cohorts is required.
Disclosures:
Alex J. Thompson - Advisory Committees or Review Panels: Gilead, Abbvie,
BMS, Merck, Spring Bank Pharmaceuticals, Arrowhead, Roche; Grant/Research
Support: Gilead, Abbvie, BMS, Merck; Speaking and Teaching: Roche, Gilead,
Abbvie, BMS
Zhaoshi Jiang - Employment: Gilead Sciences, Inc.; Stock Shareholder: Gilead
Scieces, Inc.
Dongliang Ge - Employment: Gilead Sciences, Inc
Anuj Gaggar - Employment: Gilead Sciences, Inc.
Mani Subramanian - Employment: Gilead Sciences
David B. Goldstein - Advisory Committees or Review Panels: Sever Adverse
Events Consortium; Grant/Research Support: Gilead, Astra Zeneca, Biogen,
Johnson and Johnson, Labcorp, Merck, Fundazione Telethon, NIH, CURE, UCB,
IL28B and ITPA finds
Henry Lik-Yuen Chan - Advisory Committees or Review Panels: Gilead, MSD,
Bristol-Myers Squibb, Roche, Novartis Pharmaceutical, Abbvie; Speaking and
Teaching: Echosens
Edward J. Gane - Advisory Committees or Review Panels: Novira, AbbVie, Janssen,
Gilead Sciences, Janssen Cilag, Achillion, Merck, Tekmira; Speaking and
Teaching: AbbVie, Gilead Sciences, Merck
Harry L. Janssen - Consulting: AbbVie, Bristol Myers Squibb, GSK, Gilead Sciences,
Innogenetics, Merck, Medtronic, Novartis, Roche, Janssen, Medimmune,
ISIS Pharmaceuticals, Tekmira; Grant/Research Support: AbbVie, Bristol Myers
Squibb, Gilead Sciences, Innogenetics, Merck, Medtronic, Novartis, Roche,
Janssen, Medimmune
The following authors have nothing to disclose: Ondrej Podlaha, Xin Guo, Luting
Zhuo
1652
Effect of a novel synthetic FXR agonist EYP001 on hepatitis
B virus replication in HepaRG cell line and primary
human hepatocytes
Pauline Radreau 2 , Marine Porcherot 1 , Jacky Vonderscher 2 , Vincent
Lotteau 1 , Patrice André 1 ; 1 CIRI U1111, INSERM, Lyon, France;
2 EnyoPharma, Lyon, France
HBV replication is tightly linked to bile acids (BA) metabolism.
1) BA nuclear receptor FXR binds to two response elements in
the HBV core promoter region and its activation by ligands
regulates the HBV core promoter activity 2) the Na-taurocholate
cotransporting polypeptide (NTCP) responsible of BA
uptake by hepatocytes was identified as a functional receptor
for HBV and 3) reciprocally, HBV infection induces changes
of the expression of several genes involved in the BA synthesis
pathway in the liver of infected patients. We investigated the
effect of FXR activity modulation on HBV replication by a novel
synthetic non-steroidal molecule EYP001 and compare its activity
to the BA derived 6-ethyl-chenodeoxycholic acid (6-ECDCA)
and the synthetic FXR agonist GW4064. First, we tested the
effect of EYP001 1) on the expression of genes under the control
of FXR in HepaRG cells and in primary human hepatocytes
(PHH) and 2) on activation of the membrane associated BA
receptor GpBAR1. Second, we tested the effect of these FXR
modulators on HBV replication in HepaRG and in PHH, two
cell culture models that support complete HBV replication cycle.
Cells were infected with HBV produced by the HepG2.2.15
line and treated from day 2 to 12 post infection with FXR modulators.
EYP001 up-regulates the expression of SHP mRNAs
and down-regulates the expression of the APOA1 mRNA and
thus behaves as a FXR agonist. In addition treatment for 10
days also reduces the expression of FXR mRNA, likely by an
SHP mediated negative feedback. On the opposite, EYP001
does not activate GpBAR1 and thus appears as a specific
FXR agonist. Treatment of HBV infected dHepaRG and PHH
with EYP001 as well as with 6-ECDCA or GW4064 strongly
inhibited, to similar extent, the secretion of HBV DNA, HBsAg,
HBeAg and of HBcAg synthesis in a dose dependent manner
(70 to 80 % inhibition at 1 or 10 microMol) as well as the viral
pregenomic RNA synthesis, cccDNA copies number and cellular
total HBV DNA. Cyclosporine A, an NTCP ligand and HBV
entry inhibitor, did not modify the effect of agonists suggesting
that the effect did not depend on entry inhibition. Treatment
consistently reversed the HBV induced modification of FXR and
related genes expression. In conclusion, this novel FXR agonist
led to a sustained repression of HBV replication in the HepaRG
and PHH cell culture systems. This effect was likely mediated by
the modulation of FXR activation that could perturb the complex
FXR network of transcription factors, which is highly targeted
and controlled by HBx. These data stress out the importance to
exploit drug regulation of metabolism pathways in controlling
HBV replication.
Disclosures:
Pauline Radreau - Employment: ENYO Pharma
Vincent Lotteau - Consulting: enyopharma; Stock Shareholder: enyopharma
Patrice André - Stock Shareholder: Enyo Pharma
The following authors have nothing to disclose: Marine Porcherot, Jacky Vonderscher