JMJD2D stabilises and cooperates with HBx protein to promote HBV transcription and replication

Xu Kong; Zuofeng Liu; Ruyi Zhang; Fu'an Xie; Rubing Liang; Yong Zhang; Lingling Yu; Wensheng Yang; Xi Li; Qiang Chen; Bei Li; Yilin Hong; Ming Li; Xiaogang Xia; Lingwei Gu; Lijuan Fu; Xiaohua Li; Ye Shen; Ting Wu; Chundong Yu; Wengang Li

doi:10.1016/j.jhepr.2023.100849

JMJD2D stabilises and cooperates with HBx protein to promote HBV transcription and replication

JHEP Rep. 2023 Jul 15;5(10):100849. doi: 10.1016/j.jhepr.2023.100849. eCollection 2023 Oct.

Authors

Xu Kong^{1

2}, Zuofeng Liu^{1

3}, Ruyi Zhang^{1

2}, Fu'an Xie^{1

2}, Rubing Liang^{1

2}, Yong Zhang⁴, Lingling Yu⁵, Wensheng Yang⁶, Xi Li⁷, Qiang Chen⁴, Bei Li⁴, Yilin Hong⁴, Ming Li^{1

2

8}, Xiaogang Xia², Lingwei Gu^{1

9}, Lijuan Fu¹⁰, Xiaohua Li¹¹, Ye Shen⁹, Ting Wu¹, Chundong Yu⁴, Wengang Li^{1

2}

Affiliations

¹ Cancer Research Center, School of Medicine, Xiamen University, Xiamen, China.
² Department of Hepatobiliary Surgery, Xiang'an Hospital of Xiamen University, School of Medicine, Xiamen University, Xiamen, China.
³ Department of Hepatology, Affiliated Hospital of Panzhihua University, Panzhihua, China.
⁴ State Key Laboratory of Cellular Stress Biology, Innovation Center for Cell Biology, School of Life Sciences, Xiamen University, Xiamen, China.
⁵ Department of Cardiology, Xiang'an Hospital of Xiamen University, School of Medicine, Xiamen University, China.
⁶ Department of Pathology, Chenggong Hospital of Xiamen University, Xiamen, China.
⁷ College of Arts and Sciences, Boston University, Boston, MA, USA.
⁸ Key Laboratory of Natural Medicine Research and Developing, Xiamen Medicine Research Institute, Xiamen, China.
⁹ Department of Management, Jiang Xia Blood Technology Co., Ltd., Shanghai, China.
¹⁰ Department of Infectious Diseases, Xiang'an Hospital of Xiamen University, School of Medicine, Xiamen University, Xiamen, China.
¹¹ Department of Surgery, Affiliated Fuzhou First Hospital of Fujian Medical University, Fuzhou, China.

Abstract

Background & aims: HBV infection is a global health burden. Covalently closed circular DNA (cccDNA) transcriptional regulation is a major cause of poor cure rates of chronic hepatitis B (CHB) infection. Herein, we evaluated whether targeting host factors to achieve functional silencing of cccDNA may represent a novel strategy for the treatment of HBV infection.

Methods: To evaluate the effects of Jumonji C domain-containing (JMJD2) protein subfamily JMJD2A-2D proteins on HBV replication, we used lentivirus-based RNA interference to suppress the expression of isoforms JMJD2A-2D in HBV-infected cells. JMJD2D-knockout mice were generated to obtain an HBV-injected model for in vivo experiments. Co-immunoprecipitation and ubiquitylation assays were used to detect JMJD2D-HBx interactions and HBx stability modulated by JMJD2D. Chromatin immunoprecipitation assays were performed to investigate JMJD2D-cccDNA and HBx-cccDNA interactions.

Results: Among the JMJD2 family members, JMJD2D was significantly upregulated in mouse livers and human hepatoma cells. Downregulation of JMJD2D inhibited cccDNA transcription and HBV replication. Molecularly, JMJD2D sustained HBx stability by suppressing the TRIM14-mediated ubiquitin-proteasome degradation pathway and acted as a key co-activator of HBx to augment HBV replication. The JMJD2D-targeting inhibitor, 5C-8-HQ, suppressed cccDNA transcription and HBV replication.

Conclusion: Our study clarified the mechanism by which JMJD2D regulates HBV transcription and replication and identified JMJD2D as a potential diagnostic biomarker and promising drug target against CHB, and HBV-associated hepatocarcinoma.

Impact and implications: HBV cccDNA is central to persistent infection and is a major obstacle to healing CHB. In this study, using cellular and animal HBV models, JMJD2D was found to stabilise and cooperate with HBx to augment HBV transcription and replication. This study reveals a potential novel translational target for intervention in the treatment of chronic hepatitis B infection.

Keywords: Chronic hepatitis B; HBV replication; HBx; Intervention target; JMJD2D; cccDNA transcription.