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395
Targeting Neddylation, a Potential Strategy in
Anti-HBV Therapy
Mingjie Xie1, Fan Yang2, Zhenggang Yang2, Liyan Shui2
and Min Zheng1, (1)State Key Laboratory for Diagnosis
and Treatment of Infectious Diseases, The First Affiliated
Hospital of School of Medicine, Zhejiang University, (2)State
Key Laboratory for Diagnosis and Treatment of Infectious
Diseases, The First Affiliated Hospital of School of Medicine,
Zhejiang University
Background: Hepatitis B Virus (HBV) infection remains a
global health problem, with more than 364 million chronic
carriers worldwide. Further exploration of the molecular
mechanisms in HBV replication may provide novel strategies
against HBV. Neddylation as one of post-translational
modifications plays an important role in regulating viral
infection. It is triggered by the successive activation of Nedd8-
activating enzyme E1 (NAE1). Recent studies confirmed
that HIV can utilize neddylation to induce the degradation of
host restriction factor SAMHD1H for supporting its survival.
This study aimed to investigate the role and mechanism of
neddylation in HBV life cycle and further examine the effect of
neddylation inhibition on HBV. Methods: The NAE1 inhibitor,
Pevonedistat, and siNAE1 were used to block neddylation.
HepG2.2.15, pHBV1.3-transfected HepG2 and Huh7 cells
were used for in vitro study. HBV-infected mouse model
was established by pHBV1.3 hydrodynamic injection. The
HBsAg and HBeAg levels were detected by ELISA. The
HBV DNA, mRNA and protein expression levels of interest
genes were assessed by qPCR and Western blot analysis,
respectively. Cell viability was evaluated by CCK8 assay.
Luciferase reporter assay was used to determine the activity
of promoters. Hepatic HBc expression was detected by
immunohistochemistry. Results: By using a noncytotoxic
concentration of Pevonedistat (400nM) or knocking down
NAE1 in HepG2.2.15 and pHBV1.3-transfected Huh7
and HepG2 cells, we found that Pevonedistat treatment or
NAE1 knockdown both significantly reduced HBsAg, HBeAg
and HBV DNA levels and decreased viral promoter activity
in HBV cellular models. In addition, the serum HBsAg,
HBeAg and HBV DNA were lower in HBV-infected mice by
Pevonedistat treatment as compared to those by vehicle
treatment. Furthermore, we revealed that the transcription
factor hepatocyte nuclear factor 4α (HNF4α) expression in
HBV cellular models was suppressed under Pevonedistat
treatment and the hepatic HBc and HNF4α expressions in
HBV mouse model were also decreased after Pevonedistat
treatment. Conclusion: Our results demonstrated that
neddylation blockage by NAE1 inhibition suppress HBV
replication and transcription in vitro and in vivo. Pevonedistat
may represent as a potential drug for antiviral therapy in HBV. |
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发表于 2018-10-15 08:43