Combined small molecule and loss-of-function screen uncovers estrogen receptor alpha and CAD as host factors for HDV infection and antiviral targets.

Abstract

OBJECTIVE:

Hepatitis D virus (HDV) is a circular RNA virus coinfecting hepatocytes with hepatitis B virus. Chronic hepatitis D results in severe liver disease and an increased risk of liver cancer. Efficient therapeutic approaches against HDV are absent.

DESIGN:

Here, we combined an RNAi loss-of-function and small molecule screen to uncover host-dependency factors for HDV infection.

RESULTS:

Functional screening unravelled the hypoxia-inducible factor (HIF)-signalling and insulin-resistance pathways, RNA polymerase II, glycosaminoglycan biosynthesis and the pyrimidine metabolism as virus-hepatocyte dependency networks. Validation studies in primary human hepatocytes identified the carbamoyl-phosphatesynthetase 2, aspartate transcarbamylase and dihydroorotase (CAD) enzyme and estrogen receptor alpha (encoded by ESR1) as key host factors for HDV life cycle. Mechanistic studies revealed that the two host factors are required for viral replication. Inhibition studies using N-(phosphonoacetyl)-L-aspartic acid and fulvestrant, specific CAD and ESR1 inhibitors, respectively, uncovered their impact as antiviral targets.

CONCLUSION:

The discovery of HDV host-dependency factors elucidates the pathogenesis of viral disease biology and opens therapeutic strategies for HDV cure.

• Authors: Eloi R Verrier, Amélie Weiss, Charlotte Bach, Laura Heydmann, Vincent Turon-Lagot, Arnaud Kopp, Houssein El Saghire, Emilie Crouchet, Patrick Pessaux, Thomas Garcia, Patrick Pale, Mirjam B Zeisel, Camille Sureau, Catherine Schuster, Laurent Brino, Thomas F Baumert
• Journal: Gut (Gut) Vol. 69 Issue 1 Pg. 158-167 (01 2020) ISSN: 1468-3288 [Electronic] England
• PMID: 30833451 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © Author(s) (or their employer(s)) 2020. Re-use permitted under CC BY-NC. No commercial re-use. See rights and permissions. Published by BMJ.

Chemical References

• Antiviral Agents
• CAD trifunctional enzyme
• ESR1 protein, human
• Estrogen Receptor Antagonists
• Estrogen Receptor alpha
• HIF1A protein, human
• Hypoxia-Inducible Factor 1, alpha Subunit
• Pyrimidines
• RNA, Small Interfering
• RNA, Viral
• Fulvestrant
• Aspartic Acid
• sparfosic acid
• Aspartate Carbamoyltransferase
• Dihydroorotase
• Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing)
• pyrimidine
• Phosphonoacetic Acid

Topics

• Antiviral Agents (pharmacology)
• Aspartate Carbamoyltransferase (antagonists & inhibitors, genetics, metabolism)
• Aspartic Acid (analogs & derivatives, pharmacology)
• Carbamoyl-Phosphate Synthase (Glutamine-Hydrolyzing) (antagonists & inhibitors, genetics, metabolism)
• Cell Line
• Dihydroorotase (antagonists & inhibitors, genetics, metabolism)
• Estrogen Receptor Antagonists (pharmacology)
• Estrogen Receptor alpha (antagonists & inhibitors, metabolism)
• Fulvestrant (pharmacology)
• Gene Silencing
• Hepatitis D, Chronic (drug therapy, genetics, metabolism)
• Hepatitis Delta Virus (physiology)
• Hepatocytes
• Humans
• Hypoxia-Inducible Factor 1, alpha Subunit (metabolism)
• Insulin Resistance
• Life Cycle Stages
• Loss of Function Mutation
• Phosphonoacetic Acid (analogs & derivatives, pharmacology)
• Pyrimidines (biosynthesis)
• RNA Interference
• RNA, Small Interfering (genetics)
• RNA, Viral (metabolism)
• Signal Transduction
• Virus Replication