Analysis of host factor networks during hepatitis B virus infection in primary human hepatocytes

• Published in: Virology journal 2024-08, Vol.21 (1), p.170-12, Article 170
• Main Authors: Hwangbo, Suhyun, Kim, Gahee, Choi, Yongwook, Park, Yong Kwang, Bae, Songmee, Ryu, Jae Yong, Hur, Wonhee
• Format: Article
• Language: English
• Subjects: Cells, Cultured; Gene Expression Profiling; Gene Regulatory Networks; Health aspects; Hepatitis B; Hepatitis B - genetics; Hepatitis B - virology; Hepatitis B virus; Hepatitis B virus - genetics; Hepatitis B virus - physiology; Hepatitis B, Chronic - virology; Hepatocytes - virology; Host-Pathogen Interactions; Host-virus relationships; Humans; Liver cells; Medical research; Medicine, Experimental; Physiological aspects; Primary human hepatocytes; RNA-binding proteins; Transcriptome analysis; Virus Replication
• ISSN: 1743-422X; 1743-422X
• DOI: 10.1186/s12985-024-02446-3

Chronic hepatitis B virus (HBV) infection affects around 250 million people worldwide, causing approximately 887,000 deaths annually, primarily owing to cirrhosis and hepatocellular carcinoma (HCC). The current approved treatments for chronic HBV infection, such as interferon and nucleos(t)ide analogs, have certain limitations as they cannot completely eradicate covalently closed circular DNA (cccDNA). Considering that HBV replication relies on host transcription factors, focusing on host factors in the HBV genome may provide insights into new therapeutic targets against HBV. Therefore, understanding the mechanisms underlying viral persistence and hepatocyte pathogenesis, along with the associated host factors, is crucial. In this study, we investigated novel therapeutic targets for HBV infection by identifying gene and pathway networks involved in HBV replication in primary human hepatocytes (PHHs). Importantly, our study utilized cultured primary hepatocytes, allowing transcriptomic profiling in a biologically relevant context and enabling the investigation of early HBV-mediated effects. PHHs were infected with HBV virion particles derived from HepAD38 cells at 80 HBV genome equivalents per cell (Geq/cell). For transcriptomic sequencing, PHHs were harvested 1, 2-, 3-, 5-, and 7 days post-infection (dpi). After preparing the libraries, clustering and sequencing were conducted to generate RNA-sequencing data. This data was processed using Bioinformatics tools and software to analyze DEGs and obtain statistically significant results. Furthermore, qRT-PCR was performed to validate the RNA-sequencing results, ensuring consistent findings. We observed significant alterations in the expression patterns of 149 genes from days 1 to 7 following HBV infection (R  > 0.7, q