Targeting a tolerogenic HLA-G genotype to tackle immune evasion and adaptive resistance in HBV-driven HCC.

In this study the authors intended to investigate the relationship between intrahepatic hepatitis B virus (HBV)-DNA concentrations and posthepatectomy recurrence of HBV-associated hepatocellular carcinoma (HCC). High HBV-DNA level is strongly associated with HCC development in chronic HBV infection and considered to be a risk factor of HCC recurrence. A total of 109 patients with HBV-associated HCC who underwent curative surgical resection were followed up every 3 to 6 months for a median of 82 months. Intrahepatic total HBV-DNA titer was measured in HCC and surrounding liver tissues using a TaqMan probe-based real-time polymerase chain reaction method. HBV-DNA titers in HCC and surrounding liver were compared in accordance with patients' clinical, radiologic, and histopathological characteristics. The relationships between HBV-DNA titers in HCC or surrounding liver tissues and cumulative HCC recurrence rates were determined. Of the 109 patients, 67 (62%) showed posthepatectomy recurrence of HCC. In all patients, total HBV-DNA titers were significantly higher in HCCs than in surrounding liver tissues (P=0.019). HCC recurred more frequently in patients with higher than those with lower HBV-DNA titers in surrounding liver tissues (P=0.009). In contrast, the HCC recurrence rates were similar in patients with higher and those with lower HBV-DNA titers in HCC specimens (P=0.301). Multivariate analysis showed that tumor size >5 cm (P=0.008), the presence of portal vein thrombus (P=0.001), and high HBV-DNA titer in surrounding liver tissues (P=0.002) were independent risk factors for posthepatectomy HCC recurrence in patients with HBV-associated HCC. In patients with HBV-associated HCC, high HBV-DNA titer in surrounding liver rather than in the HCC itself is associated with posthepatectomy HCC recurrence after curative surgical resection.

BackgroundNatural killer (NK) cells activation has been reported to contribute to inflammation and liver injury during hepatitis B virus (HBV) infection both in transgenic mice and in patients. However, the role of NK cells in the process of HBV-associated hepatocellular carcinoma (HCC) development has not been addressed. Killer cell immunoglobulin-like receptors (KIRs) are involved in regulating NK cell activation through recognition of specific human leukocyte antigen (HLA) class I allotypes.Methodology/Principal FindingsTo investigate whether KIR and HLA genes could influence the risk of HBV-associated HCC development, 144 HBV-infected patients with HCC and 189 well-matched HBV infectors with chronic hepatitis or cirrhosis as non-HCC controls were enrolled in this study. The presence of 12 loci of KIR was detected individually. HLA-A, -B, -C loci were genotyped with high-resolution. HLA-C group 1 homozygote (OR = 2.02; p = 0.005), HLA-Bw4-80I (OR = 2.67; p = 2.0E-04) and combination of full-length form and 22 bp-deleted form of KIR2DS4 (KIR2DS4/1D) (OR = 1.89; p = 0.017) were found associated with HCC incidence. When the combined effects of these three genetic factors were evaluated, more risk factors were observed correlating with higher odds ratios for HCC incidence (P trend = 7.4E-05). Because all the risk factors we found have been reported to result in high NK cell functional potential by previous studies, our observations suggest that NK cell activation may contribute to HBV-associated HCC development.Conclusions/SignificanceIn conclusion, this study has identified significant associations that suggest an important role for NK cells in HCC incidence in HBV-infected patients. Our study is useful for HCC surveillance and has implications for novel personalized therapy strategy development aiming at HCC prevention in HBV-infected patients.

Novel pre-mRNA splicing of intronically integrated HBV generates oncogenic chimera in hepatocellular carcinoma

Objective To explore the relationship between CXCL1 expression level and prognosis of HBV-associated hepatocellular carcinoma (HCC) after liver transplantation. Methods From 2000 to 2010, 127 recipients of liver transplantation for HBV-associated HCC in our hospital were enrolled. General hematoxylin-eosin staining and CXCL1 immunohistochemical staining were applied in formalin-fixed paraffin-embedded sections, and the Cohort′s adverse prognostic factors were analyzed retrospectively. Neutrophil migration assay and Transwell invasion assay were used to verify its mechanism. Results CXCL1 expression level is an independent prognostic factor for 5-year overall survival (OS) and disease-free survival (DFS) (P<0.05); Neutrophil migration assay and Transwell invasion assay suggested that HCC cells can secrete CXCL1 recruiting neutrophils to hepatic carcinoma tissues, and neutrophils secrete vascular endothelial growth factor A (VEGFA) promoting the invasion of HCC cells through vascular endothelial growth factor receptor 2 (VRGFR2). Conclusion High CXCL1 level in HCC tissues is an independent prognostic factor for HCC recurrence and long-term survival of patients with HBV-associated HCC after liver transplantation. CXCL1-neutrophil-VEGFA-VEGFR2 may be a mechanisms leading to HCC recurrence in patients with HBV-associated HCC after liver transplantation. Key words: Liver transplantation; Carcinoma, hepatocellular; Hepatitis B, chronic; Chemokine CXCL1

Chronic infection with hepatitis B virus (HBV) remains a significant health concern, with between 350-500 million people chronically infected worldwide. Approximately 25% of chronically infected individuals will go on to develop HBVassociated hepatocellular carcinoma (HCC), making chronic infection with HBV the leading risk factor for developing HCC. With the high incidence and mortality of HCC, it is important to fully understand the mechanisms that lead to the development of HBV-associated HCC. HBV requires a complex network of hostvirus interactions to meet its requirements for successful replication. Each of these host-virus interactions, over a decades-long chronic infection, could significantly alter the physiology of an infected hepatocyte, the target of HBV infection, and ultimately contribute to the oncogenic potential of HBV. Typically, studies of these host-virus interactions have focused on a single factor or pathway to characterize contributions to HBV replication or HBV-associated disease, but these studies have generally not considered hepatocyte physiology as a whole. We hypothesized that using broad, transcriptomic- and metabolomic -based technologies would allow us to establish a better understanding of the complexity of the host-virus interaction mediated by HBV and how an HBV infection affects overall hepatocyte physiology. To achieve this, we utilized an ex-vivo primary rat hepatocyte model and defined transcriptome-wide, HBV-mediated changes to gene expression. We also utilized metabolomic profiling to assess the impact of HBV, and the HBV X protein (HBx), on overall hepatocyte metabolism and correlated these changes to HBV-mediated changes in gene expression. Using this approach, we identified significant alterations of many genes and pathways central to hepatocyte physiology, including cell cycle regulation, lipid metabolism, and energy metabolism. Our results simultaneously identified multiple HBV-mediated changes to hepatocyte physiology, increasing our understanding of the complex relationship between HBV and an infected hepatocyte. Additionally, the identification of HBV-regulated genes will serve as the basis for future studies in understanding the physiological impact of an HBV infection. Together, these findings will allow a better understanding of HBV-mediated affects on hepatocyte physiology that could ultimately contribute to the development of HBV-associated disease, potentially guiding the generation of novel therapeutics and strategies to prevent HBV-associated HCC.

BACKGROUNDEntecavir (ETV) and tenofovir fumarate (TDF) are recommended first-line agents for the treatment of chronic hepatitis B virus (HBV) infection. However, the effect of these 2 antiviral agents on the risk for recurrence of HBV-associated hepatocellular carcinoma (HCC) after radical hepatectomy remains controversial.AIMTo compare the effect of TDF vs ETV on the risk for HCC recurrence after radical surgery for HBV-related HCC.METHODSData from consecutive patients, who received TDF or ETV after radical hepatectomy for HBV-related HCC and admitted to the Second Hospital of Longyan between December 2018 and December 2023, were retrospectively analyzed. Based on treatment method and propensity score matching (PSM), data from 100 patients were included, with 50 in each of the TDF and ETV groups, respectively. The baseline characteristics of the 2 groups were analyzed, and the risk for HCC recurrence was compared between the groups.RESULTSThe median follow-up for 100 patients [median age, 61 years; 84 male (84%)] who underwent radical resection for HBV-related HCC - Barcelona Clinic Liver Cancer stage 0 [n = 16 (16%)], stage A [n = 61 (61%)] - was 29 months (range, 12-60 months); the median tumor size was 3.0 cm (range, 2.1-4.3 cm). Sixty-eight (68%) patients exhibited HBV-DNA levels > 1000 IU/mL. Twenty-two (22%) patients tested positive for hepatitis B e antigen, in whom the HCC recurrence rate was 59.1% (13/22). After PSM, HCC recurrence rates in the ETV and TDF groups after hepatectomy were 66% (n = 33) and 42% (n = 21), respectively (P = 0.016), and cumulative recurrence rates at 1, 3, and 5 years were 26%, 58%, and 66%, and 18%, 38%, and 42%, respectively (P = 0.045).CONCLUSIONTDF treatment is associated with a lower risk for HCC-related outcomes than that for ETV in patients with HBV-associated HCC after curative therapy.

BackgroundOur previous report suggested that centrosomal P4.1-associated protein (CPAP) is required for Hepatitis B virus (HBV) encoded non-structure protein X (HBx)-mediated nuclear factor kappa light chain enhancer of activated B cells (NF-κB) activation. CPAP is overexpressed in HBV-associated hepatocellular carcinoma (HCC); however, the interaction between CPAP and HBx in HBV-HCC remains unclear.MethodsThe mRNA expression of CPAP and HBx was analyzed by quantitative-PCR (Q-PCR). NF-κB transcriptional activity and CPAP promoter activity were determined using a reporter assay in Huh7 and Hep3B cells. Immunoprecipitation (IP) and in situ proximal ligation assay (PLA) were performed to detect the interaction between CPAP and HBx. Chromatin-IP was used to detect the association of cAMP response element binding protein (CREB) and HBx with the CPAP promoter. Cell proliferation was measured using cell counting kit CCK-8, Bromodeoxyuridine (5-bromo-2′-deoxyuridine, BrdU) incorporation, and clonogenic assays. The tumorigenic effects of CPAP were determined using xenograft animal models.ResultsHBx can transcriptionally up-regulate CPAP via interacting with CREB. Overexpressed CPAP directly interacted with HBx to promote HBx-mediated cell proliferation and migration; SUMO modification of CPAP was involved in interacting with HBx. Knocked-down expression of CPAP decreased the HBx-mediated tumorigenic effects, including cytokines secretion. Interestingly, overexpressed CPAP maintained the HBx protein stability in an NF-κB-dependent manner; and the expression levels of CPAP and HBx were positively correlated with the activation status of NF-κB in HCC. Increased expression of CPAP and CREB mRNAs existed in the high-risk group with a lower survival rate in HBV-HCC.ConclusionThe interaction between CPAP and HBx can provide a microenvironment to facilitate HCC development via enhancing NF-κB activation, inflammatory cytokine production, and cancer malignancies. This study not only sheds light on the role of CPAP in HBV-associated HCC, but also provides CPAP as a potential target for blocking the hyper-activated NF-κB in HCC.

HBV related liver disease ranges from hepatitis, cirrhosis, and hepatocellular carcinoma (HCC), which has a 5-year survival rate of 14% because it is difficult to diagnose at its curative stages. Chronic hepatitis B virus (HBV) infections is the major etiology of HCC, associated with over 50% of HCC cases worldwide and up to 70-80% in endemic areas. 85-90% of HBV related HCCs (HBV-HCC) contain integrated HBV DNA. During chronic HBV infection, HBV DNA has been shown to integrate at random sites in the host chromosome creating a unique genetic signature in each HBV infected hepatocytes. This makes the pool of HBV DNA integration sites highly complex in the liver. This genetic signature in HBV-infected liver along with the fundamental mechanism of carcinogenesis, which is clonal expansion, would result in HCC tissue containing only one to few clonally expanded integration sites, resulting in reduced complexity. Thus, we hypothesize the detection of reduced complexity of HBV DNA integration sites in the circulation can be a potential biomarker for HBV-HCC. In order to study the complexity of cell free circulating HBV DNA integration sites in the circulation, a targeted next generation sequencing (NGS) assay for urine DNA was developed. We chose to use urine in order to avoid detecting of HBV DNA from infectious viruses. Targeted enrichment was performed using an in-solution hybridization platform with biotinylated RNA baits covering the entire HBV genome. The assay was tested using library constructed DNA controls: Hep3B, which contains integrated DNA, and HepG2, which contains no HBV DNA to confirm targeted capture of HBV DNA. Next, matching tissue and urine DNA and DNA from urine collected after surgical removal of HCC tumor from seven HBV-HCC patients were collected and underwent library construction. The library DNA samples were subsequently enriched with HBV and HCC-specific RNA baits. Enriched library DNA was sequenced on Illumina's MiSeq platform. Although the analysis is still ongoing, we have successfully analyzed the complexity of cell-free circulating HBV DNA integration sites in urine. We detected 46 integration sites in the tissue and urine where the location of integrated HBV DNA mapped primarily (89%) to a known HBV integration breakpoint region, HBV DR1-2. Furthermore in 3 HBV-HCC patients, a pattern of reduced complexity of cell-free circulating HBV DNA integration sites was found in urine before the removal of HCC, as compared to urine collected after surgery suggesting reduced complexity of HBV DNA integration sites in HBV-HCC patients. A similar NGS analysis of the urine of hepatitis and cirrhosis patients is in progress. Overall, if successful this study will demonstrate the potential of detecting reduced complexity of HBV DNA integration sites as a biomarker for HBV-HCC screening and provide insight to the involvement of HBV DNA integration sites in HBV-HCC carcinogenesis. Citation Format: Selena Lin, Surbhi Jain, Batbold Boldbaatar, Timothy Block, Wei Song, Ying-Hsiu Su. Comprehensive analysis of the complexity of HBV DNA integration sites in the circulation of patients with HBV-related liver disease. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 876. doi:10.1158/1538-7445.AM2014-876

PurposeEarly diagnostic biomarkers of hepatocellular carcinoma (HCC) are needed to distinguish hepatitis B virus (HBV) associated HCC (HBV-HCC) patients from at-risk patients. We assessed the diagnostic values of serum Integrin beta-like 1 (ITGBL1) for early-stage HBV-HCC.Patients and MethodsWe recruited 716 participators including 299 in the training and 417 in the validation stage, (HBV-HCC, chronic hepatitis B (CHB), HBV‐related liver cirrhosis (HBV-LC), and healthy controls) between 2017 and 2020 from three centers. Serum ITGBL1 was measured by ELISA. Receiver operating characteristic (ROC) was used to calculate diagnostic accuracy.ResultsThe serum levels of ITGBL1 in HBV-HCC patients were significantly lower than those in CHB and HBV-LC patients. This result was confirmed in the follow-up patients who progressed from HBV-LC to HCC. The optimum diagnostic cutoff value of serum ITGBL1 was 47.93ng/mL for detection of early-stage HBV-HCC. The serum ITGBL1 has higher diagnostic accuracy than AFP20 in differentiating the early-stage HBV-HCC from the at-risk patients (area under curve [AUC] 0.787 vs 0.638, p<0.05). For AFP-negative (<20ng/mL) HBV-HCC patients, serum ITGBL1 maintained diagnostic accuracy (training cohort: AUC 0.756, 95% confidence interval [CI] 0.683–0.819, sensitivity 68.18%, and specificity 68.85%; validation cohort: 0.744, 0.686–0.796, 81.13%, and 55.88%). Combination ITGBL1 with AFP20 significantly increased diagnostic accuracy in differentiating the HBV-HCC from at-risk patients (AUC 0.840; 0.868) than ITGBL1 (AUC 0.773, p<0.05; 0.732, p<0.0001) or AFP20 (AUC 0.705, p<0.0001; 0.773, p<0.0001) alone.ConclusionThe serum level of ITGBL1 improved identification of AFP-negative HBV-HCC patients, and increased diagnostic accuracy with AFP20 together in the early detection of HBV-HCC.

BackgroundLiver cancer is the sixth most common cancer worldwide and the third leading cause of cancer-related death. As a chronic liver disease, many studies have shown that the immune response plays a key role in the progression of liver cancer. Chronic hepatitis B virus (HBV) infection is one of the high-risk factors for HCC, accounting for 50%–80% of HCC cases worldwide, and little is known about the immune status of HBV associated hepatocellular carcinoma (HBV-HCC), therefore, we aimed to explore the changes in peripheral immunity in patients with HBV-HCC.MethodsIn this study, patients with HBV-HCC (n=26), patients with hepatitis B-related cirrhosis (HBV-LC) (n=31) and healthy volunteers (n=49) were included. The lymphocytes and their subpopulation phenotypes in peripheral blood were characterized. In addition, we explored the effect of viral replication on peripheral immunity in patients with HCC and analyzed the circulating immunophenotypic characteristics at different stages of HCC with flow cytometry.ResultsFirstly, our results showed that the percentages of total αβ T cells in the peripheral blood of HBV-HCC patients was significantly decreased compared to healthy subjects. Secondly, we found that naïve CD4+ T cells in HBV-HCC patients were significantly reduced, terminally differentiated CD8+ T cells, homing memory CD8+ T cells and Th2 cells were increased in peripheral circulation in HBV-HCC patients. Moreover, in the peripheral blood of HBV-HCC patients, expression of TIGIT on CD4+ T cells and PD-1 on the surface of Vδ 1 T cells was increased. In addition, we found that sustained viral replication resulted in up-regulation of TIM3 expression on CD4+ T cells, and TIM3+ γδ T cells increased in peripheral circulation in patients with advanced HBV-HCC.ConclusionOur study showed that circulating lymphocytes in HBV-HCC patients exhibited features of immune exhaustion, especially in HCC patients with persistent viral replication and in patients with intermediate and advanced HBV-HCC, including decreased frequency of T cells and elevated expression of inhibitory receptors including TIGIT and TIM3 on CD4+ T cells and γδ T cells. Meanwhile, our research suggests that the combination of CD3+ T cell and CD8+HLADR+CD38+ T cell may be a potential diagnostic indicator for HBV-HCC. These findings could help us to better understand the immune characteristics of HBV-HCC and explore the immune mechanisms and immunotherapy strategies for HBV-HCC.

To evaluate the reliability of serum human phosphorylated exospindle polar-like proteinase 1 (ESPL1) as a serum biomarker for early diagnosis of hepatitis B virus (HBV)-related hepatocellular carcinoma (HBV-HCC). This retrospective study was conducted on 266 patients with chronic hepatitis B (CHB), liver cirrhosis (LC), and HBV-related HCC. Data on demographics and clinical information were collected, and ESPL1 levels were measured using enzyme linked immunosorbent assay. Levels of ESPL1, alpha-fetoprotein (AFP), and protein induced by vitamin K absence -II (PIVKA-II) were compared at different disease stages, and spearman correlation analysis was used to assess their relationship with clinical markers. The diagnostic accuracy of ESPL1, AFP, and PIVKA-II for early HBV- HCC was assessed using ROC curve analysis. The study comprised 121 patients diagnosed with CHB, 98 patients with LC, and 47 patients with HBV-HCC. Serum ESPL1 levels show an increasing trend across groups with chronic HBV infection, CHB, LC, and HBV-HCC, with levels at 224.6 ng/L, 285.8 ng/L, and 440.4 ng/L (in pairwise comparison, P<0.05). Serum AFP and PIVKA-II levels displayed no significant statistical differences between the CHB and LC groups. Spearman correlation analysis revealed that levels of ESPL1, PIVKA-II, and AFP are not influenced by clinical characteristics and show no correlation with each other. ROC curve analysis indicated that the optimal diagnostic threshold for ESPL1 in HBV-HCC is 345.7 ng/L, with AUC values for ESPL1, PIVKA-II, and AFP being 0.797 (95% CI: [0.708-0.886]), 0.788 (95% CI: [0.718-0.858]), and 0.572 (95% CI: [0.523-0.624]). In AFP and PIVKA-II negative patients, the AUC values for ESPL1 diagnosis of HBV-HCC were 0.79 and 0.83. ESPL1 is a potential biomarker for tracking chronic HBV infection and predicting the development of HBV-HCC. Monitoring ESPL1 levels in serum could help with early detection and personalized screening HBV-HCC for individuals with chronic HBV infection.

Most hepatocellular carcinoma (HCC) patients worldwide do not receive curative treatments. Alternative treatments for most HCC patients include palliative treatments, such as transarterial chemoembolization (TACE), chemotherapy, and radiotherapy. Although statins may be a chemopreventive treatment option for reducing hepatitis B virus (HBV)- and hepatitis C virus (HCV)-related HCC risks, their therapeutic effects are unknown. This study evaluated the effects of statin on HCC patients receiving palliative treatment.Data from the National Health Insurance claims database and cancer registry databases of The Collaboration Center of Health Information Application, Taiwan, were analyzed. We included HCC patients who were treated between January 1, 2001, and December 31, 2010, and followed them from the index date to December 31, 2012. The inclusion criteria were presence of HBV carrier-related HCC, age >20 years, and having received TACE, radiotherapy, or chemotherapy as palliative treatment. The exclusion criteria were cancer diagnosis before HCC was confirmed, surgery, liver transplantation, radiofrequency ablation, or percutaneous ethanol injection as curative treatment, missing sex-related information, HCC diagnosis before HBV, and age <20 years. We enrolled 20,200 HCC patients.The median follow-up duration was 1.66 years (interquartile range, 0.81). In total, 1988 and 18,212 patients received palliative treatment with and without statin use, respectively. HCC patients who received palliative treatment with statin use had lower HCC-specific deaths in all stages than those who received palliative treatment without statin use (P = 0.0001, 0.0002, 0.0012, and 0.0002, and relative risk (RR) = 0.763, 0.775, 0.839, and 0.718, for stages I–IV, respectively). In all-cause and HCC-specific deaths, decreasing trends (P for trend <0.0001) of adjusted hazard ratios (aHRs) were observed in all stages with no treatment, statin use only, palliative treatment only, and palliative treatment plus statin use. The aHRs of all-cause and HCC-specific deaths increased with the progress in cancer stage and reduced with the use of advanced therapeutic modalities (P for trend <0.0001). Differences in HBV- and non-HBV-related HCC were solely due to statin use. Statin use alone reduced HCC-specific deaths by 36% in non-HBV-related HCC in stage I and 50% in HBV-related HCC in stages II and III. With a relatively substantial reduction in mortality, the therapeutic effects of statin use were stronger in HBV-related HCC than in non-HBV-related HCC.Palliative treatments are critical for HCC patients. Multiple therapeutic methods with statin use reduced the mortality risk. Statins prolong the survival of patients with advanced HCC receiving palliative treatment, thus demonstrating its therapeutic value as an adjuvant treatment. Furthermore, statin-based palliative treatment in early stage HCC remarkably reduced the number of deaths. For patients who cannot tolerate palliative treatments, statin use only might possibly reduce mortality, particularly in HBV-related early stage HCC patients (>50% reduction in HCC deaths).

Metabolic reprogramming is a hallmark of cancer. The nicotinamide phosphoribosyltransferase (NAMPT)-mediated salvage pathway maintains sufficient cellular NAD levels and is required for tumorigenesis and development. However, the molecular mechanism by which NAMPT contributes to HBV-associated hepatocellular carcinoma (HCC) remains not fully understood. In the present study, our results showed that NAMPT protein was obviously upregulated in HBV-positive HCC tissues compared with HBV-negative HCC tissues. NAMPT was positively associated with aggressive HCC phenotypes and poor prognosis in HBV-positive HCC patients. NAMPT overexpression strengthened the proliferative, migratory, and invasive capacities of HBV-associated HCC cells, while NAMPT-insufficient HCC cells exhibited decreased growth and mobility. Mechanistically, we demonstrated that NAMPT activated SREBP1 (sterol regulatory element-binding protein 1) by increasing the expression and nuclear translocation of SREBP1, leading to the transcription of SREBP1 downstream lipogenesis-related genes and the production of intracellular lipids and cholesterol. Altogether, our data uncovered an important molecular mechanism by which NAMPT promoted HBV-induced HCC progression through the activation of SREBP1-triggered lipid metabolism reprogramming and suggested NAMPT as a promising prognostic biomarker and therapeutic target for HBV-associated HCC patients.

Exploring predictive biomarkers and therapeutic strategies of ICBs has become an urgent need in clinical practice. Increasing evidence has shown that ARID1A deficiency might play a critical role in sculpting tumor environments in various tumors and might be used as pan-cancer biomarkers for immunotherapy outcomes. The current study aims to explored the immune-modulating role of ARID1A deficiency in Hepatitis B virus (HBV) related hepatocellular carcinoma (HBV-HCC) and its potential immunotherapeutic implications. In the current study, we performed a comprehensive analysis using bioinformatics approaches and pre-clinical experiments to evaluate the ARID1A regulatory role on the biological behavior, and immune landscape of Hepatitis B virus (HBV) related hepatocellular carcinoma (HBV-HCC). A total of 425 HBV-related hepatocellular carcinoma patients from TCGA-LIHC, AMC and CHCC-HBV cohort were enrolled in bioinformatics analysis.Immunohistochemical staining of HBV-HCC specimens and ARID1A deficiency cellular models were used to validate the results of the analysis. Our results have shown that ARID1A deficiency promoted tumor proliferation and metastasis. More importantly, ARID1A deficiency in HBV-HCC was associated with the higher TMB, elevated immune activity, and up-regulated expression of immune checkpoint proteins, especially TIM-3 in HBV-HCC. Further, the expression of Galectin-9, which is the ligand of TIM-3, was elevated in the ARID1A knockout HBV positive cell line. To conclude, we have shown that the ARID1A deficiency was correlated with more active immune signatures and higher expression of immune checkpoints in HBV-HCC. Additionally, the present study provides insights to explore the possibility of the predictive role of ARID1A in HBV-HCC patients responsive to immunotherapy.

Aims:To investigate the feasibility of serum extra spindle pole bodies-like 1 (ESPL1) used as a biomarker for patients with hepatitis B virus (HBV)-related hepatocellular carcinoma (HCC).Methods:131 chronic HBV-infection patients were recruited and divided into HBV S gene integration, non-HBV S gene integration, chronic hepatitis B (CHB), HBV-related liver cirrhosis (LC) and HBV-related HCC group, 24 non-HBV-related HCC patients were selected as HCC control group, 30 people without HBV-infection as healthy control group. Serum ESPL1 were detected and compared.Results:ESPL1 level of integration group was significantly higher than that of non-integration group (346.7 vs 199.6 ng/ml, P = 0.000) and healthy control group (346.7 vs 41.3 ng/ml, P = 0.000). ESPL1 level of non-integration group was significantly higher than that of healthy control group (199.6 vs 41.3 ng/ml, P = 0.000); ESPL1 levels in chronic HBV-infection related groups were increased in turn according to CHB group (95.8 ng/ml), HBV-related LC group (268.2 ng/ml), HBV-related HCC group (279.9 ng/ml) and integration group (346.7 ng/ml). Except that there was no significant difference in ESPL1 levels between HBV-related LC and HCC group (P = 0.662), pairwise comparisons between other groups showed significant differences (P < 0.05). ESPL1 level of HBV-related HCC group was significantly higher than that of non-HBV-related HCC group (279.9 vs 46.6 ng/ml, P = 0.000), there was no noticeable difference between non-HBV-related HCC and healthy control group (46.6 vs 41.3 ng/ml, P = 0.848). ESPL1 level of HBV-related HCC group after resection was significantly lower than that of before resection (178.4 vs 260.8 ng/ml, P = 0.000).Conclusions:Chronic HBV-infection patients with high ESPL1 level may indicate HBV S gene integration and is a high-risk population for HBV-related HCC. Serum ESPL1 can be used as a biomarker for screening HBV-related HCC high-risk population and monitoring recurrence.