Association of TRIM29 with HBV replication and the antiviral effect of pegylated interferon α-2b
10.12449/JCH241111
- VernacularTitle:三结构域蛋白29(TRIM29)与HBV复制及聚乙二醇干扰素α-2b抗病毒作用的关联性分析
- Author:
Xin LIAO
1
;
Baofang ZHANG
Author Information
1. 贵州医科大学临床医学院,贵阳 550000
- Keywords:
Hepatitis B Virus;
Tripartite Motif-Containing Protein 29;
PEG-IFNα;
Signaling Pathway
- From:
Journal of Clinical Hepatology
2024;40(11):2191-2200
- CountryChina
- Language:Chinese
-
Abstract:
Objective To preliminarily investigate the association of TRIM29 with HBV replication and the antiviral effect of pegylated interferon α-2b(PEG-IFN-α-2b),since TRIM29 protein is involved in the development and progression of a variety of diseases and is closely associated with the replication of some DNA and RNA viruses.Methods A total of 64 chronic hepatitis B(CHB)patients who attended the outpatient service of Department of Infectious Diseases,The Affiliated Hospital of Guizhou Medical University,from October 2021 to June 2022 were enrolled,among whom there were 34 treatment-na?ve patients and 30 patients treated with PEG-IFN-α-2b,and 30 healthy volunteers in Physical Examination Center were enrolled as controls.Related data were collected,including age,sex,alanine aminotransferase,aspartate aminotransferase,total bilirubin,direct bilirubin,HBV DNA,and peripheral blood mononuclear cells(PBMCs).HepG2 and HepG2.2.15 cells were used as cell models and were transfected with TRIM29-specific overexpressed plasmid or siRNA and control plasmid.HepG2 cells and Huh7 cells were treated with PEG-IFN-α-2b(0,10,100,1 000,and 10 000 U/mL),and HepG2.2.15 cells were treated with TRIM29-specific siRNA or negative control combined with PEG-IFN-α-2b.ELISA was used to measure the concentrations of HBsAg and HBeAg;qRT-PCR was used to measure the relative expression levels of TRIM29 and HBV RNA;Western blot was used to measure the protein expression levels of STING,p-TBK1,TBK1,pIRF3,IRF3,MX1,and IFIT1;co-immunoprecipitation assay was used to observe the interaction between TRIM29 and STING protein.The independent-samples t test was used for comparison of normally distributed continuous data between two groups,and a one-way analysis of variance was used for comparison between multiple groups,with the least significant difference t-test for further comparison between two groups;the chi-square test or the Fisher's exact test was used for comparison of categorical data between two groups.Results The CHB patients had a significantly higher expression level of TRIM29 in peripheral blood than the healthy controls(P<0.001).In cell experiments,the expression levels of HBsAg,HBeAg,and HBV RNA increased with the upregulation of TRIM29 expression and decreased with downregulation of TRIM29 expression(P<0.05).TRIM29 bound to STING and degraded STING via protease,and compared with the control group,there were no significant changes in the total protein levels of TBK1 and IRF3 after overexpression of TRIM29,while there were significant reductions in the expression levels of STING,p-TBK1,and p-IRF3(P<0.05).The protein and mRNA expression levels of TRIM29 decreased with the increase in the concentration of PEG-IFN-α-2b for the treatment of HepG2 and Huh7 cells(P<0.01).During the treatment with PEG-IFN-α-2b,the CHB patients had a gradual reduction in the mRNA expression level of TRIM29,and there was a significant difference between the early response group and the non-response group(P<0.05).In the context of treatment with an equal volume of PEG-IFN-α-2b,compared with the control group,there were significant increases in the protein expression levels of Mx1 and IFIT1 in HepG2.2.15 cells after TRIM29 knockdown(P<0.05).There was a gradual reduction in the expression of TRIM29 in CHB patients during the early stage of PEG-IFN-α-2b treatment.Conclusion TRIM29 targets and degrades STING and promotes HBV replication by inhibiting the STING-TBK1-IRF3 signaling pathway.TRIM29 interferes with the antiviral effect of PEG-IFN-α-2b,and the expression level of TRIM29 in PBMCs of CHB patients may be used as an indicator for predicting the response of patients to PEG-IFN-α-2b therapy.
