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Local ablation for hepatocellular carcinoma, a non-surgical option that directly targets and destroys tumor cells, has advanced significantly since the 1990s. Therapies with different energy sources, such as radiofrequency ablation, microwave ablation, and cryoablation, employ different mechanisms to induce tumor necrosis. The precision, safety, and effectiveness of these therapies have increased with advances in guiding technologies and device improvements. Consequently, local ablation has become the first-line treatment for early-stage hepatocellular carcinoma. The lack of organized evidence and expert opinions regarding patient selection, pre-procedure preparation, procedural methods, swift post-treatment evaluation, and follow-up has resulted in clinicians following varied practices. Therefore, an expert consensus-based practical recommendation for local ablation was developed by a group of experts in radiology and hepatology from the Research Committee of the Korean Liver Cancer Association in collaboration with the Korean Society of Image-Guided Tumor Ablation to provide useful information and guidance for performing local ablation and for the pre- and post-treatment management of patients. [ABSTRACT FROM AUTHOR]

Background/Aims: Young Korean men are obligated to serve in the military for 18 to 21 months. We investigated the effects of military service on steatotic liver disease (SLD) and other metabolic parameters. Methods: Pre-enlistment health check-up performed from 2019 to 2022 and in-service health check-up performed from 2020 to 2022 were merged as paired data. SLD was defined as a hepatic steatosis index of 36 or higher. Hypertension (HTN) and hypertriglyceridemia were also included in the analysis. Results: A total of 503,136 paired cases were included in the analysis. Comparing pre-enlistment and in-service health check-ups, the prevalence of SLD (22.2% vs 17.6%, p<0.001), HTN (7.6% vs 4.3%, p<0.001), and hypertriglyceridemia (8.1% vs 2.9%, p<0.001) decreased during military service. In terms of body mass index, the proportion of underweight (8.2% vs 1.4%, p<0.001) and severely obese (6.1% vs 4.9%, p<0.001) individuals decreased over time. Regarding factors associated with SLD development and resolution, age was positively associated with SLD development (odds ratio, 1.146; p<0.001) and a health check-up interval of <450 days was a protective factor for SLD development (odds ratio, 0.746; p<0.001). Those serving in the marines were less likely to develop SLD, whereas those serving in the navy were more likely to develop SLD. Serving in the army or the navy was negatively associated with SLD resolution, whereas serving in the air force was positively associated with SLD resolution. Conclusions: The prevalence of SLD, HTN, and hypertriglyceridemia decreased substantially during Korean military service. [ABSTRACT FROM AUTHOR]

Background/Aims: Bile duct invasion (BDI) is rarely observed in patients with advanced hepatocellular carcinoma (HCC), leading to hyperbilirubinemia. However, the efficacy of pretreatment biliary drainage for HCC patients with BDI and obstructive jaundice is currently unclear. Thus, the aim of this study was to assess the effect of biliary drainage on the prognosis of these patients. Methods: We retrospectively enrolled a total of 200 HCC patients with BDI from multicenter cohorts. Patients without obstructive jaundice (n=99) and those who did not undergo HCC treatment (n=37) were excluded from further analysis. Finally, 64 patients with obstructive jaundice (43 subjected to drainage and 21 not subjected to drainage) were included. Propensity score matching was then conducted. Results: The biliary drainage group showed longer overall survival (median 10.13 months vs 4.43 months, p=0.004) and progression-free survival durations (median 7.00 months vs 1.97 months, p<0.001) than the non-drainage group. Multivariate analysis showed that biliary drainage was a significantly favorable prognostic factor for overall survival (hazard ratio, 0.42; p=0.006) and progression-free survival (hazard ratio, 0.30; p<0.001). Furthermore, in the evaluation of first response after HCC treatment, biliary drainage was beneficial (p=0.005). Remarkably, the durations of overall survival (p=0.032) and progression-free survival (p=0.004) were similar after propensity score matching. Conclusions: Biliary drainage is an independent favorable prognostic factor for HCC patients with BDI and obstructive jaundice. Therefore, biliary drainage should be contemplated in the treatment of advanced HCC with BDI to improve survival outcomes [ABSTRACT FROM AUTHOR]

Background/Aims Despite its prognostic impact, nutritional status has not yet been integrated into the assessment ofhepatocellular carcinoma (HCC). This study investigated the association between geriatric nutritional risk index (GNRI) and overall survival (OS) in patientswith HCC using a nationwide registry. Methods Data from the Korea Central Cancer Registry between 2008 and 2019 were analyzed. We explored the integration of the GNRI with the albumin–bilirubin (ALBI) grade for prognostic stratification. Restricted cubic spline regression was used to assess the association between GNRI and survival, stratified by ALBI grade. Results Among the 16,416 treatment-naïve HCC patients, the ALBI grades were distributed as follows: grade 1, 7,409; grade 2,7,445; and grade 3, 1,562. Patients were categorized according to Barcelona Clinic Liver Cancer (BCLC) stages: 5,132 stage 0/A, 2,608 stage B, 5,289 stageC, and 968 stage D. The median OS for all patients was 3.1 years (95% CI:3.0–3.2) and significantly differed with the inclusion of ALBI grade and GNRI (Figure 1; p<0.001). The effect of combining ALBI grade and GNRI was further evaluated for each BCLC stage. This risk stratification showed a significant correlation with OS for each BCLC stage (all p<0.001), except for stage D (p=0.082). Multivariate analysis revealed that a combination of favorable ALBI grade and high GNRI score was independently associated with decreased mortality risk. Conclusion The GNRI was significantly correlated with OS across ALBI grades and BCLC stages. Integrating the GNRI into the ALBI grade may enhance risk stratification for patients with HCC. [ABSTRACT FROM AUTHOR]

Background/Aims PD-L1 expression is abundant not only in malignant cells but also in infiltrating cells within the tumor microenvironment (TME) of hepatocellular carcinoma (HCC). This study explored the association between PD-L1 expression in TME and outcomes in HCC patients treated with atezolizumab plus bevacizumab (AB), emphasizing the implications of PD-L1 expression in both malignant and tumor-infiltrating cells. Methods This retrospective study included 72 patients with HCC who underwent percutaneous core needle liver biopsy before AB treatment between September 2020 and December 2023. PD-L1 expression on tumor tissues was assessed using the combined positive score (CPS) with cutoff values of 1 and 10, utilizing antibody clone 22C3 (Dako). Results The distribution of PD-L1 CPS included 24 patients with CPS <1, 33 patients with CPS 1–10, and 15 patients with CPS ≥10. Significant differences in OS were observed across the three groups, with CPS ≥10 showing the highest survival rates (p=0.010). Patients with CPS ≥10 had better OS than those with CPS <10 (mOS 14.8 vs. 8.3 months, p=0.046), and CPS ≥1 had better OS than CPS <1 (p=0.021). For PFS, the CPS ≥10 group had the highest mPFS of 11.0 months among the three groups (p=0.044). Objective response rates (ORR) were higher in the PD-L1 CPS ≥10 group than in the 1-10 and <1 group (53.3%, 27.3%, and 16.7%, respectively; p=.047). Multivariate analysis identified that PD-L1 expression ≥10 and ≥1 were associated with favorable outcomes regarding OS (hazard ratio [HR] 0.283, p=.027 and HR 0.303, p=.006, respectively). Conclusion Higher PD-L1 expression in malignant and tumor-infiltrating cells was associated with better OS, PFS, and ORR in patients with HCC treated with AB. [ABSTRACT FROM AUTHOR]

The article discusses the use of clinical parameters as predictive factors for the treatment of hepatocellular carcinoma (HCC) with atezolizumab and bevacizumab. The study found that factors such as albumin-bilirubin grade, age, C-reactive protein (CRP), and alpha-fetoprotein (AFP) levels were significant predictors of progression-free survival (PFS). The CRAFITY score, which combines AFP and CRP levels, was shown to be an independent prognostic factor. The study confirms the efficacy of atezolizumab and bevacizumab for the treatment of advanced HCC and highlights the importance of clinical parameters in guiding personalized treatment decisions. [Extracted from the article]

Background/Aims Metabolic dysfunction-associated steatotic liver disease (MASLD) is a growing health issue, with activated fibroblasts playing a key role in its progression. This study evaluated the potential of nintedanib, a drug used for idiopathic pulmonary fibrosis (IPF), in treating MASLD by targeting these fibroblasts. Additionally, the study explored how cancer-associated fibroblasts (CAFs) in hepatocellular carcinoma (HCC) are influenced by Immunoglobulin A (IgA) complexes, which may alter immune responses and contribute to inflammation. Methods Fibroblasts from MASLD patients were treated with nintedanib and compared to sorafenib in vitro. A mouse model of fibrosis was used for in vivo analysis. Fibroblasts, including CAFs, were also isolated from HCC patients and treated with IgA dimers to assess their impact on CD71 and PD-L1 expression. The effects of nintedanib on these IgA-stimulated fibroblasts were evaluated. Single-cell sequencing identified HMGN3 as a key regulator of matrix CAFs, with subsequent experiments assessing nintedanib's effect on HMGN3 expression. Results Nintedanib significantly reduced activated fibroblast viability in vitro, outperforming sorafenib. In the mouse model, nintedanib selectively eliminated FAP+PD-L1+ fibroblasts, suggesting its therapeutic potential for MASLD. CAFs expressed higher levels of α-smooth muscle actin and fibroblast activation protein than non-tumor fibroblasts. IgA treatment increased PD-L1 expression on CAFs and reduced TNF-α levels in CD8+ T cells, indicating a possible suppression of antitumor immunity. Nintedanib effectively eliminated these IgA-stimulated fibroblasts. Additionally, nintedanib reduced HMGN3 expression in hepatic stellate cells. Conclusion Nintedanib shows promise as a treatment for MASLD by targeting and eliminating activated fibroblasts. Its ability to remove IgA-stimulated fibroblasts suggests it could also be beneficial in managing fibrosis in liver diseases like HCC. [ABSTRACT FROM AUTHOR]

Background/Aims It has been 4 years since the FDA approved the immunotherapybased regimen atezolizumab-bevacizumab as the first-line treatment of hepatocellular carcinoma. Hence, accurate biomarkers predicting prognosis of the relatively new treatment has not been discovered. In 2022, Scheiner et al suggested a scoring system named CRAFITY score, including CRP and AFP, that accurately predicted prognosis of HCC patients who were treated with atezolizumab-bevacizumab combination therapy. There have also been multiple studies suggesting PIVKA-II as an additionally accurate biomarker and NLR(neutrophil-lymphocyte ratio) as an accurate predictive factor for HCC patients. Naturally, we experimented in including PIVKA-II and NLR into the CRAFITY score system. Methods We included 146 patients diagnosed with HCC who were treated with atezolizumab-bevacizumab. We assigned 1 point for having an AFP ≥100 ng/ml and 1 point for having a CRP ≥1 mg/dl. As we added the PIVKA-II and NLR as new factors included in the scoring system (mCRAFITY), we assigned 1 point for having a PIVKA-II ≥40 mAU/ml and an additional 1 point for having NLR >2.5. We used the AUROC(area under the receiver operating characteristic) to evaluate the performance of our scoring system. Results The AUROC curve showed that the mCRAFITY score had better performance at 6 months and 12 months but worse performance at 18 months in predicting the prognosis of HCC patients. When OS and PFS was calculated, patients with mCRAFITY scores of less than 3 had significantly better prognosis than those with scores more than or equal to 3. (p value=0.0015 and 0.0391 respectively) Conclusion In conclusion, the mCRAFITY score that includes PIVKA-II and NLR may be more accurate than the previously announced CRAFITY score. Patients with mCRAFITY scores less than 3 had significantly better prognosis than whom at scores more than or equal to 3. [ABSTRACT FROM AUTHOR]

Background/Aims The CD69+ tissue-resident T-cell population comprises a considerable proportion of intrahepatic T cells and provides an early, rapid adaptive immune response. However, the characteristics of CD69- populations and their role need to be elucidated. Methods We collected background liver tissues, which were obtained during hepatectomy in patients with various etiologies of chronic liver disease. The patient cohort included n=28 with the following etiologies : hepatitis B virus (HBV, 46%), alcohol-related liver disease (21%), HBV combined with alcoho-related liver disease (HBV+Alc, 29%), and unknown etilogy (4%). Single-cell dissociation and flow cytometry were performed to analyze the liver-infiltrating T-cell population. Fatty changes and fibrosis grade were confirmed by histologic examinations. Results The CD69- subpopulation was more enriched in CD4+ T cells compared to CD8+ T cells. CD69- CD4+ /CD8+ T cells showed a similar activation phenotype but expressed PD-1 less, compared to the CD69+ subpopulation. Interestingly, the CD69- group exhibited higher ex vivo expression of perforin and granzyme B in CD4+ T cells. While perforin expression was lower in CD69- CD4+ T cells compared to CD69- /CD69+ CD8+ T cells, granzyme B levels were similar to those in CD69+ CD8+ T cells, with CD69+ CD8+ T cells having a greater granzyme B+ population. NKG2D+ and CD56+ populations among CD69- CD4+ T cells had more perforin or granzyme B, suggesting they might be activated in an innate-like manner. The frequency of cytotoxic CD69- CD4+ T cells was positively correlated with the serum ALT level but was negatively correlated with liver fibrosis. Conclusion We have identified for the first time that cytotoxic CD4+ T cells among liver-infiltrating T cells are mainly CD69- population, which can recirculate. This population might be associated with liver inflammation, which needs to be further validated. [ABSTRACT FROM AUTHOR]

Background/Aims Daratumumab, a monoclonal antibody targeting CD38, is currently used as a first-line therapy in patients with multiple myeloma. This study aims to investigate the risk of HBR in patients with the anti-HBc antibody and to evaluate the efficacy of prophylactic antiviral therapy in this population. Methods Data from 345 multiple myeloma patients treated with daratumumab between 2014 and 2024 were retrospectively collected. Patients with the anti-HBc antibody were included in the study, while those with positive HBsAg prior to daratumumab treatment were excluded. HBR was defined as HBsAg seroconversion or detection of HBV DNA until subsequent chemotherapy or 12 months after cessation of daratumumab if no subsequent chemotherapy was performed. Results Among the 345 patients, 109 had the anti-HBc antibody and were included in the analysis. The median duration of daratumumab treatment was 101 days, during which ten patients received prophylactic antiviral therapy (9 with entecavir and 1 with tenofovir alafenamide). Overall, 9 patients experienced HBR, with 3 suffering from severe hepatitis (ALT exceeding 200U/L) due to HBR. No patient who received prophylactic antiviral therapy experienced HBR. The cumulative incidence rates of HBR in the non-antiviral treatment group were 8.0% at 12 months after the start of daratumumab. When restricted mean survival time (RMST) was employed to compare the area under the Kaplan-Meier curve of HBR-free survival in both groups, the antiviral treatment group was significantly higher than the non-antiviral treatment group (Difference in RMST=2.00, p=0.002). HBsAb titers less than 100 IU/L (odds ratio 7.527, p=0.057) and anti-HBc antibody titer (odds ratio 1.234, p=0.156) showed a tendency towards an increased incidence of HBR. Conclusion There is a considerable risk of HBR in multiple myeloma patients with the anti-HBc antibody undergoing daratumumab treatment. The use of antiviral therapy may prevent HBR in these at-risk patients. [ABSTRACT FROM AUTHOR]

Background/Aims: According to the results of several studies, the outcome of hepatitis C virus (HCV) reactivation is not as severe as the outcome of hepatitis B virus reactivation. The aim of this study was to evaluate the effect of pharmacological immunosuppression on HCV reactivation. Methods: The medical records of patients who underwent systemic chemotherapy, corticosteroid therapy, or other immunosuppressive therapies between January 2008 and March 2015 were reviewed. Subsequently, 202 patients who were seropositive for the anti-HCV antibody were enrolled. Exclusion criteria were: unavailability of data on HCV RNA levels, a history of treatment for chronic hepatitis C, and the presence of liver diseases other than a chronic HCV infection. Results: Among the 120 patients enrolled in this study, hepatitis was present in 46 patients (38%). None of the patients were diagnosed with severe hepatitis. Enhanced replication of HCV was noted in nine (27%) of the 33 patients who had data available on both basal and follow-up HCV RNA loads. Reappearance of the HCV RNA from an undetectable state did not occur after treatment. The cumulative rate of enhanced HCV replication was 23% at 1 year and 30% at 2 years. Conclusions: Although enhanced HCV replication is relatively common in HCV-infected patients treated with chemotherapy or immunosuppressive therapy, it does not lead to serious sequelae. [ABSTRACT FROM AUTHOR]

Background/Aims In this study, we aimed to identify the amount and patterns of immune cell infiltration in autoimmune hepatitis (AIH), and to compare them with those of drug-induced liver injury (DILI). Methods From March 2016 to December 2018, 15 patients with AIH were prospectively enrolled in this study. For comparison, 22 patients with DILI in the same period were analyzed in the same way. Liver biopsy was performed, and immunohistochemical stain for CD3, CD68, CD20, and CD38 was done. For some patients, immune cells were harvested from fresh liver biopsy samples, and multicolor flow cytometry was used for the immunophenotyping of the infiltrated immune cells. Results First, we identified that activated CD8 and CD4 T cells were more infiltrated in the livers from patients with AIH than those from healthy controls by multicolor flow cytometry. The amounts of T cells, macrophages, and B cells infiltration had no associations with serum aspartate aminotransferase, alanine aminotransferase (ALT), alkaline phosphatase or gamma glutamyltranspeptidase in patients with AIH. Furthermore, serum levels of IgG and IgA were not correlated with the amounts of immune cells infiltrations, either. Next, we compared the infiltration of the immune cell subsets between patients with AIH and those with DILI. Baseline serum ALT was significantly higher in patients with DILI, but the levels of activated T cells (CD3+CD38+) infiltration were not significantly different between AIH and DILI. However, a larger number of macrophages were infiltrated in the livers in DILI than those in AIH, suggesting that drug-induced injury of hepatocytes triggers innate immunity more vigorously. Conclusions Both patients with AIH and DILI have activated T cells and macrophages infiltrated in the injured liver. Higher ALT and more macrophages infiltration in DILI suggest that drug-induced injury of hepatocytes might trigger innate immunity more vigorously than autoimmunity-mediated mechanisms. [ABSTRACT FROM AUTHOR]

Background/Aims Drug-induced liver injury (DILI) is liver injury caused by the interplay between drugs, their metabolites and host immune response. The aim of this study is to characterize liver infiltrating immune cells in DILI and to evaluate the correlation between infiltrating immune cells and clinical outcome. Methods From January 2017 to February 2019, 23 patients with DILI were prospectively enrolled in this study. Diagnosis of DILI was based on patient medication history after exclusion of other etiology (virus, alcohol, autoimmune, ischemic hepatitis or extrahepatic obstruction of the bile duct). Liver biopsy was performed, and immunohistochemical stain for CD3, CD68, CD20, and CD38 was done. Experienced pathologist confirmed the pathologic and immunohistochemical findings. Results The causes included health foods or dietary supplements (n=8, 34.8%), folk remedies (n=6, 26.1%), medications (n=4, 17.4%), herb (n=3, 13.0%). Eighteen patients (78.3%) treated with steroid. Two patients progressed to hepatic failure and recovered after steroid treatment. All patients were completely recovered from DILI at 1 month after diagnosis, whether they were treated with steroid or not. The frequency of infiltrated T cells, macrophages, and B cells was quantified by immunohistochemistry. The amounts of T cells, showed linearly ascending correlation with total bilirubin (r=0.370, p=0.030) and Model for End-Stage Liver Disease (MELD) score (r=0.340, p=0.047). The amounts of macrophages showed linearly ascending correlation with total bilirubin (r=0.401, p=0.020), aspartate aminotransferase (r=0.445, p=0.010), and MELD score (r=0.402, p=0.020). Conclusions In this study, we found the positive correlation between the amounts of T cells and macrophages infiltrations and deterioration of liver function in DILI. Favorable responses to steroid therapy suggest that vigorous innate and adaptive immune responses play critical roles in DILI. [ABSTRACT FROM AUTHOR]

Background/Aims In this study, we have demonstrated that microRNA99a (miR-99a), a microRNAs (miRNA) that has been reported to regulate mammalian target of rapamycin (mTOR) pathway and cell survival, restricts hepatitis C virus replication by blocking mTORC1-mediated de novo lipogenesis. Methods Huh-7 cells were infected by cell culture-produced hepatitis C virus (HCV; JFH-1; genotype 2a). HCVcc infected cells and full-genomic HCV replicon (FGR) cells were used for miRNA experiments. Serum samples of HCV-infected patients and those of age-matched healthy controls were collected and miR99a level was quantified by real-time qPCR. Results The relative expression of miR-99a was significantly lower in the sera of the patients with chronic HCV infection compared to the sera of those without viral hepatitis. HCVcc infection caused downregulation of miR-99a expression in Huh-7 cells, and FGR cells showed lower expression of miR-99a than wild-type Huh-7 cells. Forced expression of miR-99a-5p resulted in dramatic decrease in intracellular and secreted HCV RNA levels in HCVcc-infected cells. Using in silico analysis tools, mTOR was identified as a potential target of miR99a with a high binding score. Transfection of miR-99a-5p mimics resulted in the decreased level of mTOR protein and decreased phosphorylation of 4E-BP and S6K in FGR cells. The mRNA and protein level of sterol regulatory element binding protein-1c (SREBP-1c), the master transcriptional regulator of fatty acid and triglyceride synthesis, was decreased when either miR99a mimics or si-mTOR was transfected. In cells transfected with miR-99a-5p mimics, oleic acid-induced intracellular lipid accumulation were significantly decreased compared to cells treated with scrambled miRNAs. Forced expression of mTOR rescued the replication of HCV RNA and lipid droplet accumulation in miR99a mimics-transfected FGR cells, suggesting that mTOR is responsible for the anti- lipogenic activity of miR-99a. Conclusions Our data clearly demonstrate that miR-99a ameliorate intracellular lipid accumulation by regulating the expression of SREBP-1c and its target genes, and cause inefficient replication and packaging of intracellular HCV. [ABSTRACT FROM AUTHOR]

Background/Aims Although the therapeutic mechanisms of bone marrow-derived mesenchymal stem cells (BM-MSCs) are still unclear, BM-MSCs play key therapeutic roles in liver fibrosis. Furthermore, microRNAs (miRNAs) are regulators in hepatic differentiation and liver fibrosis. miR-101-3p is upregulated during hepatic trans-differentiation, whereas miR-101-3p is downregulated in a stage of liver fibrosis. The purpose of this study is to investigate miR-101-3p's roles in the hepatic differentiation of human BM-MSC and hepatic stellate cell (HSC) activation. Methods To detect the miRNAs, the next-generation sequencing (NGS) was performed in hBM-MSCs before and after hepatic differentiation, normal and fibrotic liver tissues. miRTarBase and miRDB were used to predict targets of miR-101-3p. hBM-MSC was treated with mimic, inhibitor, or siRNAduring the hepatic differentiation. hHSC LX2 was treated with transforming growth factor beta 1 (TGF-β1), and with or without miR-101-3p mimics, inhibitors, and EZH2 siRNA. The role of miR-101-3p was identified through the change of liver-specific genes, epithelial-mesenchymal transition markers, and fibrosis genes using the quantitative real-time PCR (qRT-PCR) and western blotting. Results As a result of NGS analysis, miR-101-3p showed a higher expression level in differentiated hepatocyte-like cells than hBM-MSC but lower expression level in liver fibrosis than the normal liver. miR-101-3p caused the increase in the liver-specific genes in hBM-MSC, while mesenchymal markers, fibrosis markers, and apoptosis inhibitor genes decreased in LX2. At this time, the expression of EZH2 was decreased and the same result as miR-101-3p overexpression was obtained by knockdown of EZH2. As a result, miR-101-3p mimic promoted the hepatic differentiation of hBM-MSC and inhibited the TGF-β1 mediated LX2 activation via regulating EZH2. Conclusions In this study, we identified miR-101-3p that can regulate the hepatic differentiation of hBM-MSC and the hepatic fibrosis by targeting EZH2. Our results demonstrate that miR-101-3p may be a biomarker, monitoring the response to therapeutic effect by BM-MSC in liver fibrosis. [ABSTRACT FROM AUTHOR]

Background/Aims While it has long been known that hepatitis B virus (HBV) integrates into the host genome, the biological role of HBV integration in hepatocarcinogenesis still remains uncertain. The study aimed to localize HBV integrants throughout the host genome by high-throughput sequencing method. Methods Next-generation sequencing (NGS)-based target enrichment sequencing was performed to detect HBV integration in paired tumor/non-tumor tissues from 33 hepatocellular carcinoma (HCC) patients. The Illumina NGS workflow was conducted in the following steps: gDNA sonication to generate DNA libraries, HBV sequence capture using HBV probes, followed by high-throughput sequencing. Chimeric count >10 and AVG_MQ >20 was regarded as true signal. Results HBV integration was identified in all (25/25, 100%) HBsAg-positive and two (2/8, 25%) HBsAg-negative patients with HCC. Two patients who lost HBsAg before HCC development also harbored HBV integration. The average number of integration sites in HBV-related HCC and adjacent non-tumor tissues was 12.45 and 24.04 per sample, respectively. HBV integration most frequently occurred in chromosomes 5 and 10. Recurrent target genes for HBV integration included TERT, MLL4, and PREX2 for tumor and FN1 for non-tumor. The major hotspot breakpoints of host and HBV genomes were located at upstream of hTERT gene and nt 1,600-1,800 of HBV genome, respectively. Integrations in tumor were over-presented in promoter or exon and less presented in intergenic region. Gene-annotation analysis indicated that the recurrent HBV integrations are enriched in cancer-associated genes. HBV integration breakpoints were validated by Sanger sequencing in twelve randomly selected tumor tissues. Conclusions Our study reports a new NGS-based high-throughput targeted sequencing with high detection ability to identify HBV integration in the human genome. This cost-effective method facilitates a survey of HBV integration in a large number of samples in an unbiased way and helps in investigating the oncogenic role of HBV integration in HCC. [ABSTRACT FROM AUTHOR]