Combined small molecule and loss-of-function screen uncovers estrogen receptor alpha and CAD as host factors for HDV infection and antiviral targets
Résumé
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.
Mots clés
antiviral therapy
hepatitis D
liver
screening
Antiviral therapy
Hepatitis D
Screening
Liver HDV: hepatitis D virus
HBV: hepatitis B virus
HSPG: heparan sulfate proteoglycan
NTCP: sodium taurocholate co-transporting polypeptide
HD-Ag: delta antigen
RNP: ribonucleoprotein
EBV: Epstein-Barr virus
CAD: carbamoyl-phosphate synthetase 2
aspartate transcarbamylase
and dihydroorotase
ESR1: estrogen receptor 1
PHH: primary human hepatocytes
DHODH: dihydroorotate dehydrogenase
PALA: N-(Phosphonoacetyl)-L-aspartic acid
TFV: tenofovir
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