Identification of a Potential miRNA-mRNA Regulatory Network Associated With the Prognosis of HBV-ACLF

Shanshan Ma; Zhongyang Xie; Lingjian Zhang; Ya Yang; He Jiang; Xiaoxi Ouyang; Yalei Zhao; Qiuhong Liu; Xiaowei Xu; Lanjuan Li

doi:10.3389/fmolb.2021.657631

Identification of a Potential miRNA-mRNA Regulatory Network Associated With the Prognosis of HBV-ACLF

Front Mol Biosci. 2021 Apr 28:8:657631. doi: 10.3389/fmolb.2021.657631. eCollection 2021.

Authors

Shanshan Ma^{1

2

3}, Zhongyang Xie^{1

2

3}, Lingjian Zhang^{1

2

3}, Ya Yang^{1

2

3}, He Jiang^{1

2

3}, Xiaoxi Ouyang^{1

2

3}, Yalei Zhao^{1

2

3}, Qiuhong Liu^{1

2

3}, Xiaowei Xu^{3

4}, Lanjuan Li^{1

2

3}

Affiliations

¹ State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China.
² National Clinical Research Center for Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China.
³ Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China.
⁴ Department of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou, China.

Abstract

Background: Hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF) is a life-threatening disease with a high mortality rate; the systemic inflammatory response plays a vital role in disease progression. We aimed to determine if a miRNA-mRNA co-regulatory network exists in the peripheral blood mononuclear cells (PBMCs) of HBV-ACLF patients, which might be important for prognosis.

Methods: In patients with HBV-ACLF meeting COSSH-ACLF criteria, age, liver cirrhosis and INR were independent risk factors for 28-day and 90-day poor prognosis. COSSH-ACLFs was a superior prognostic model. mir-6840-3p-JADE2 may promote the progression of ACLF and lead to poor prognosis. Meanwhile, mir-6840-3p and mir-6861-3p can be used as markers of short-term poor prognosis. Finally, ALSS treatment is not only the blood material exchange of patients, but also changes part of the immune state of patients. Among them, cytokine cytokine receptor interaction may play an important role in determining the therapeutic effect.

Methods: Transcriptome-wide microRNA (miRNA) and mRNA microarrays were used to define the miRNA and mRNA expression profiles of the PBMCs of HBV-ACLF patients in a discovery cohort. The targets of the miRNAs were predicted. We built a miRNA-mRNA regulatory network through bioinformatics analysis, and used quantitative real-time polymerase chain reaction (qRT-PCR) to assess the importance of candidate miRNAs and mRNAs. We also assessed the direct and transcriptional regulatory effects of miRNAs on target mRNAs using a dual-luciferase reporter assay.

Results: The miRNA/mRNA PBMC expression profiles of the discovery cohort, of whom eight survived and eight died, revealed a prognostic interactive network involving 38 miRNAs and 313 mRNAs; this was constructed by identifying the target genes of the miRNAs. We validated the expression data in another cohort, of whom 43 survived and 35 died; miR-6840-3p, miR-6861-3p, JADE2, and NR3C2 were of particular interest. The levels of miR-6840-3p and miR-6861-3p were significantly increased in the PBMCs of the patients who died, and thus predicted prognosis (areas under the curve values = 0.665 and 0.700, respectively). The dual-luciferase reporter assay indicated that miR-6840-3p directly targeted JADE2.

Conclusion: We identified a prognostic miRNA-mRNA co-regulatory network in the PBMCs of HBV-ACLF patients. miR-6840-3p-JADE2 is a potential miRNA-mRNA pair contributing to a poor prognosis.

Keywords: acute-on-chronic liver failure; hepatitis B virus; miRNA; network; prognosis.