HBV-Integration Studies in the Clinic: Role in the Natural History of Infection

Hepatitis B virus (HBV) infection is a major global health problem causing acute and chronic liver disease that can lead to liver cirrhosis and hepatocellular carcinoma (HCC). HBV covalently closed circular DNA (cccDNA) is essential for viral replication and the establishment of a persistent infection. Integrated HBV DNA represents another stable form of viral DNA regularly observed in the livers of infected patients. HBV DNA integration into the host genome occurs early after HBV infection. It is a common occurrence during the HBV life cycle, and it has been detected in all the phases of chronic infection. HBV DNA integration has long been considered to be the main contributor to liver tumorigenesis. The recent development of highly sensitive detection methods and research models has led to the clarification of some molecular and pathogenic aspects of HBV integration. Though HBV integration does not lead to replication-competent transcripts, it can act as a stable source of viral RNA and proteins, which may contribute in determining HBV-specific T-cell exhaustion and favoring virus persistence. The relationship between HBV DNA integration and the immune response in the liver microenvironment might be closely related to the development and progression of HBV-related diseases. While many new antiviral agents aimed at cccDNA elimination or silencing have been developed, integrated HBV DNA remains a difficult therapeutic challenge.

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Functional effects of HBV-DNA integration in peripheral blood mononuclear cells from subjects with chronic HBV and HDV/HBV infection

Journal of Hepatology ◽

10.1016/s0168-8278(88)80173-9 ◽

1988 ◽

Vol 7 ◽

pp. S56

Keyword(s):

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Hbv Dna ◽

Hbv Infection ◽

Peripheral Blood Mononuclear ◽

Dna Integration ◽

Blood Mononuclear Cells ◽

Chronic Hbv ◽

Hbv Dna Integration

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Hepatitis B Virus DNA Integration Occurs Early in the Viral Life Cycle in anIn VitroInfection Model via Sodium Taurocholate Cotransporting Polypeptide-Dependent Uptake of Enveloped Virus Particles

Journal of Virology ◽

10.1128/jvi.02007-17 ◽

2018 ◽

Vol 92 (11) ◽

pp. e02007-17 ◽

Cited By ~ 38

Author(s):

Thomas Tu ◽

Magdalena A. Budzinska ◽

Florian W. R. Vondran ◽

Nicholas A. Shackel ◽

Stephan Urban

Keyword(s):

Hepatitis B Virus ◽

Liver Cancer ◽

Hepatitis B ◽

Chronic Infection ◽

Hbv Dna ◽

Hbv Infection ◽

Dna Integration ◽

B Virus ◽

Hbv Dna Integration

ABSTRACTChronic infection by hepatitis B virus (HBV) is the major contributor to liver disease worldwide. Though HBV replicates via a nuclear episomal DNA (covalently closed circular DNA [cccDNA]), integration of HBV DNA into the host cell genome is regularly observed in the liver in infected patients. While reported as a prooncogenic alteration, the mechanism(s) and timing of HBV DNA integration are not well understood, chiefly due to the lack ofin vitroinfection models that have detectable integration events. In this study, we have established anin vitrosystem in which integration can be reliably detected following HBV infection. We measured HBV DNA integration using inverse nested PCR in primary human hepatocytes, HepaRG-NTCP, HepG2-NTCP, and Huh7-NTCP cells after HBV infection. Integration was detected in all cell types at a rate of >1 per 10,000 cells, with the most consistent detection in Huh7-NTCP cells. The integration rate remained stable between 3 and 9 days postinfection. HBV DNA integration was efficiently blocked by treatment with a 200 nM concentration of the HBV entry inhibitor Myrcludex B, but not with 10 μM tenofovir, 100 U of interferon alpha, or a 1 μM concentration of the capsid assembly inhibitor GLS4. This suggests that integration of HBV DNA occurs immediately after infection of hepatocytes and is likely independent ofde novoHBV genome replication in this model. Site analysis revealed that HBV DNA integrations were distributed over the entire human genome. Further, integrated HBV DNA sequences were consistent with double-stranded linear HBV DNA being the major precursor. Thus, we have established anin vitrosystem to interrogate the mechanisms of HBV DNA integration.IMPORTANCEHepatitis B virus (HBV) is a common blood-borne pathogen and, following a chronic infection, can cause liver cancer and liver cirrhosis. Integration of HBV DNA into the host genome occurs in all known members of theHepadnaviridaefamily, despite this form not being necessary for viral replication. HBV DNA integration has been reported to drive liver cancer formation and persistence of virus infection. However, when and the mechanism(s) by which HBV DNA integration occurs are not clear. In this study, we have developed and characterized anin vitrosystem to reliably detect HBV DNA integrations that result from a true HBV infection event and that closely resemble those found in patient tissues. Using this model, we showed that integration occurs when the infection is first established. Importantly, we provide here a system to analyze molecular factors involved in HBV integration, which can be used to develop strategies to halt its formation.

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Hepatitis B virus (HBV) DNA integration in patients with occult HBV infection and hepatocellular carcinoma

Liver International ◽

10.1111/liv.12807 ◽

2015 ◽

Vol 35 (10) ◽

pp. 2311-2317 ◽

Cited By ~ 58

Author(s):

Carlo Saitta ◽

Gianluca Tripodi ◽

Adalberto Barbera ◽

Antonio Bertuccio ◽

Antonina Smedile ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Hepatitis B Virus ◽

Hepatitis B ◽

Hbv Dna ◽

Hbv Infection ◽

Occult Hbv Infection ◽

Dna Integration ◽

Occult Hbv ◽

B Virus ◽

Hbv Dna Integration

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OC-8 Analysis and characterization of HBV DNA integration in patients with occult HBV infection and hepatocellular carcinoma

Digestive and Liver Disease ◽

10.1016/s1590-8658(11)60009-1 ◽

2011 ◽

Vol 43 ◽

pp. S67

Author(s):

C. Saitta ◽

T. Pollicino ◽

G. Raffa ◽

A. Bertuccio ◽

A. Barbera ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Hbv Dna ◽

Hbv Infection ◽

Occult Hbv Infection ◽

Dna Integration ◽

Occult Hbv ◽

Hbv Dna Integration

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HBV-human DNA integration in non-hepatic tumors.

Journal of Clinical Oncology ◽

10.1200/jco.2020.38.15_suppl.e13544 ◽

2020 ◽

Vol 38 (15_suppl) ◽

pp. e13544-e13544

Author(s):

Lei Liu ◽

Ming Liu ◽

Tanxiao Huang ◽

Wenting Liao ◽

Mimi Fu ◽

...

Keyword(s):

Cancer Patients ◽

Tumor Cells ◽

Tumor Tissue ◽

Hbv Dna ◽

Hbv Infection ◽

Subtype C ◽

Dna Integration ◽

Subtype B ◽

Genome Integration ◽

Hbv Dna Integration

e13544 Background: Hepatitis B virus (HBV) infection in hepatocytes may induce chronic liver damage, which further leads to hepatocellular carcinoma. HBV infection was also reported in other cell types including peripheral blood mononuclear cells (PBMCs). Moreover, epidemiological evidence suggested that HBV infection was associated with various types of cancers. In this study, we investigated whether HBV DNA integration was observed in extrahepatic tumor cells. Methods: The study enrolled thousands of Chinese patients with different types of cancers. Peripheral blood and tumor tissue samples were collected, plasma cfDNA, PBMC gDNA and tumor tissue DNA were extracted. By targeted capture and next-generation sequencing, we sequenced the cancer-related genes and identified HBV DNA integration events in these samples. Moreover, the subtype of HBV captured in the host genome was verified, and the ratio of reads mapped to the HBV genome was calculated. Results: Samples with more than 0.1% of the sequencing reads mapped to the HBV genome were identified as HBV positive cases. Thirty-two liver cancer patients were subtype-b HBV positive, among which 8 tumor samples exhibited HBV genome integration. Three colorectal cancer patients were infected by subtype-c HBV, only one tumor sample carried HBV DNA integration. Two lung cancer patients were shown to be HBV positive; one was infected by subtype-b HBV and the other carried subtype-c HBV. Subtype-b and-c HBV genome was observed in the tumor sample of two cholangiocarcinoma patients, whereas subtype-b HBV DNA was identified in the PBMC of one gastrointestinal stromal cancer patient. Conclusions: HBV genome integration was identified in the tumor cells of lung, colorectal, biliary ducts, and gastrointestinal stromal cancer. The mechanisms by which the HBV genome integrates into these tissues requires further investigation.

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Molecular Diagnosis in Differentiating Active and Inactive Forms of Hepatitis B Virus Carriers

Journal of University of Babylon for Pure and Applied Sciences ◽

10.29196/jubpas.v26i10.1838 ◽

2018 ◽

Vol 26 (10) ◽

pp. 41-45

Author(s):

Salah Tofik Jalal Balaky ◽

Saeed Ghulam Hussain ◽

Amer Ali Khaleel ◽

Furat Tahseen Sabeer ◽

Ahang Hasan Mawlood

Keyword(s):

Nucleic Acid ◽

Hepatitis B ◽

Cross Sectional Study ◽

Hbv Dna ◽

Hbv Infection ◽

Detection Methods ◽

Rt Pcr ◽

Cross Sectional ◽

Blood Samples ◽

Acid Test

Background & objectives: Introducing a nucleic acid test program is aimed to diagnose and reduces the risk of viral infection or transmission. DNA assay for HBV can detect infection in the windows period, chronic occult infection and can discriminate between active and inactive HBV infection. This cross-sectional study designed to diagnose, analyze HBV infection and to differentiate active from inactive infection based on viral DNA detection. Methods: Blood samples were collected from 256 patients previously diagnosed on the clinical ground as hepatitis B seropositive in Erbil Central Lab. The viral nucleic acid quantitative assessment was done for the collected samples using RT-PCR. Q-square was performed for statistical analysis. Results: Out of 256 collected blood samples 93 (36.3%) showed HBV-DNA positive titers above 50 IU/ml. Among positive subjects, 67 (72.04%) was categorized as inactive carriers (˂ 2000-20.000 IU/ml HBV-DNA titers). Conclusions: The data produced from this study confirmed the importance of the RT-PCR technique in sensitivity and reliability as a superior diagnostics tool specifically in differentiating active from inactive HBV carriers.

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