HBx-induced degradation of Smc5/6 complex impairs homologous recombination-mediated repair of damaged DNA

• Published in: Journal of hepatology Vol. 76; no. 1; pp. 53 - 62
• Main Authors: Sekiba, Kazuma, Otsuka, Motoyuki, Funato, Kazuyoshi, Miyakawa, Yu, Tanaka, Eri, Seimiya, Takahiro, Yamagami, Mari, Tsutsumi, Takeya, Okushin, Kazuya, Miyakawa, Kei, Ryo, Akihide, Koike, Kazuhiko
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 01.01.2022; Elsevier Science Ltd
• Subjects: Animals; Carcinoma, Hepatocellular - complications; Carcinoma, Hepatocellular - pathology; cccDNA; Cell Cycle Proteins - adverse effects; Chromosomal Proteins, Non-Histone - adverse effects; Chromosome 5; CRISPR; DDB1; Degradation; Deoxyribonucleic acid; Disease Models, Animal; DNA; DNA damage; DNA repair; Genetic transformation; HBx; Hepatocellular carcinoma; Homologous recombination; Liver - drug effects; Liver - pathology; Liver Neoplasms - complications; Liver Neoplasms - pathology; Mice; Nitazoxanide; Recombinational DNA Repair - drug effects; Recombinational DNA Repair - immunology; Statistics, Nonparametric; Trans-Activators - drug effects; Transcription; Transgenic mice; Viral Regulatory and Accessory Proteins - drug effects; DDB1; Nitazoxanide; cccDNA; DNA damage; HBx
• ISSN: 0168-8278; 1600-0641; 1600-0641
• DOI: 10.1016/j.jhep.2021.08.010

HBV causes hepatocellular carcinoma (HCC). While it was recently shown that the ability of HBV X protein (HBx) to impair the Smc5/6 (structural maintenance of chromosome 5/6) complex is important for viral transcription, HBx is also a potent driver of HCC. However, the mechanism by which HBx expression induces hepatocarcinogenesis is unclear. Degradation of the Smc5/6 complex and accumulation of DNA damage were observed in both in vivo and in vitro HBV infection models. Rescue experiments were performed using nitazoxanide (NTZ), which inhibits degradation of the Smc5/6 complex by HBx. HBx-triggered degradation of the Smc5/6 complex causes impaired homologous recombination (HR) repair of DNA double-strand breaks (DSBs), leading to cellular transformation. We found that DNA damage accumulated in the liver tissue of HBV-infected humanized chimeric mice, HBx-transgenic mice, and human tissues. HBx suppressed the HR repair of DSBs, including that induced by the CRISPR-Cas9 system, in an Smc5/6-dependent manner, which was rescued by restoring the Smc5/6 complex. NTZ restored HR repair in, and colony formation by, HBx-expressing cells. Degradation of the Smc5/6 complex by HBx increases viral transcription and promotes cellular transformation by impairing HR repair of DSBs. The hepatitis B virus expresses a regulatory protein called HBV X protein (or HBx). This protein degrades the Smc5/6 complex in human hepatocytes, which is essential for viral replication. We found that this process also plays a key role in the accumulation of DNA damage, which contributes to HBx-mediated tumorigenesis. [Display omitted] •DNA damage accumulated in HBV-infected and HBx-expressing liver tissues.•Smc5/6 degradation by HBx impaired homologous recombination repair, driving tumorigenesis.•Restoration of Smc5/6 by nitazoxanide rescued homologous recombination repair and inhibited tumorigenesis.