Cause Of Hepatocellular Carcinoma Research Articles

Impact of metformin on clinical outcomes in advanced hepatocellular carcinoma treated with immune checkpoint inhibitors

AbstractBackground and AimsNon‐alcoholic steatohepatitis (NASH) is a common cause of hepatocellular carcinoma (HCC) worldwide. Emerging data suggests NASH‐induced HCC could be associated with less response to immune checkpoint inhibitor (ICI)‐based therapy. Metformin has been associated with improved outcomes in cancers like melanoma treated with ICIs, but its impact on HCC is not well defined. The purpose of this study was to examine the effect of metformin on clinical outcomes in patients with advanced HCC treated with ICIs.MethodsWe retrospectively analysed patients with advanced HCC treated with ICIs in first and later‐line settings between 2015 and 2021. The primary endpoints were overall survival (OS), progression‐free survival (PFS), and objective response rate (ORR) as assessed according to Response Evaluation Criteria in Solid Tumours (RECIST) version 1.1. Patients were stratified based on their usage of metformin.ResultsOur study included 18 patients in the metformin group and 93 patients in the non‐metformin group. The most common causes of HCC were viral hepatitis (52%), NASH (29%), and alcohol (8%). ORR was 5.6% in the metformin group vs 22.6% in the non‐metformin group (P = .0987). Median OS was 10.8 months versus 45.9 months (HR = 1.99, 95% CI = 0.95–4.21, P = .065) and median PFS was 2.5 months versus 6.6 months (P = .077) in the metformin and non‐metformin groups, respectively. Regardless of metformin usage, OS was significantly worse in patients with poor ECOG performance status, HCC aetiology of NASH, MELD score 10–23, AFP >= 400, and use of ICIs in later lines of therapy.ConclusionsMetformin use was associated with a trend, although not statistically significant, toward a worse ORR, OS and PFS in advanced HCC patients treated with ICIs.

Perspectives on the Underlying Etiology of HCC and Its Effects on Treatment Outcomes.

Hepatocellular carcinoma (HCC) continues to be a serious medical problem with poor prognosis worldwide. The distribution of the major etiologies of HCC is changing due to the progress of anti-viral treatments, including hepatitis B virus (HBV) suppression by nucleoside/nucleotide analogues (NAs) and increased sustained virologic response (SVR) rates by direct-acting antivirals (DAAs) for hepatitis C virus (HCV), as well as the rising trend of nonviral liver disease. Although viral hepatitis remains the most common cause of HCC, non-alcoholic liver disease (NAFLD) with metabolic syndrome and alcohol-associated liver disease (ALD) are increasing. Effective and well-tolerated NAs treatment can slow the disease progression of chronic HBV infection to cirrhosis, end-stage liver disease, and reduce HCC risk. Treatment with NAs is also associated with significant improvement in the long-term survival of patients with HBV infection who already have HCC. DAAs have achieved viral elimination in almost all patients with HCV without significant adverse events, even in patients with decompensated liver cirrhosis and HCC. Similarly, DAA therapy can reduce disease progression, liver and non-liver complications, and improve the long-term survival of patients with chronic HCV infection with or without HCC. Meanwhile, NAFLD is a rapidly increasing cause of HCC along with the epidemics of obesity and type 2 diabetes globally. NAFLD-related HCC can occur in patients without cirrhosis and is known to have a lower survival rate than viral hepatitis-related HCC. Since there is currently no specific pharmacotherapy effective for NAFLD, lifestyle modification and prevention of complications are important to improve prognosis. Additionally, ALD is the second fastest-growing cause of HCC-related deaths, especially with an accelerated trend since the COVID-19 pandemic. This review provides an overview of the epidemiologic trends in the etiologies of HCC, and the progress of treatments for each etiology and the impact on outcome in the patients with HCC.

Response to antiviral therapy for chronic hepatitis C and risk of hepatocellular carcinoma occurrence in Japan: a systematic review and meta-analysis of observational studies

In Japan, hepatocellular carcinoma (HCC) is a leading cause of cancer mortality and hepatitis C virus infection is a major cause of HCC. We conducted a systematic review and meta-analysis of published studies evaluating patient response to antiviral therapy for chronic hepatitis C on the risk of HCC occurrence in Japan. Articles were searched using terms determined a priori through PubMed, screened by title and abstract, and selected by full-text assessment according to criteria determined a priori, including HCC occurrence in response to interferon (IFN)-based or IFN-free therapy, Japanese study, and 2 or more years of follow-up. We excluded studies on HCC recurrence. We calculated the pooled estimate of the crude incidence rate ratio with data from the selected studies using the person-years method with Poisson regression model and pooled estimate of the hazard ratio adjusted for potential confounders reported by the studies using a random effects model. A total of 26 studies were identified, all of which examined only IFN-based therapy as a result of the selection process. The pooled estimate (95% confidence interval [CI]) of 25 studies was 0.37 (0.33–0.43) for sustained virologic response (SVR) and 1.70 (1.61–1.80) for non-SVR for the HCC incidence rate per 100 person-years, and 0.22 (0.19–0.26) for the incidence rate ratio (SVR vs. non-SVR). The pooled estimate of the hazard ratio (95% CI) of HCC incidence adjusted for potential confounders of 8 studies was 0.25 (0.19–0.34). SVR to interferon therapy for chronic hepatitis C reduces the risk of HCC occurrence.

Resection of NAFLD/NASH-related Hepatocellular Carcinoma (HCC): Clinical Features and Outcomes Compared with HCC Due to Other Etiologies.

Non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH) are the leading causes of hepatocellular carcinoma (HCC) worldwide. Limited data exist on surgical outcomes for NAFLD/NASH-related HCC compared with other HCC etiologies. We evaluated differences in clinicopathological characteristics and outcomes of patients undergoing surgical resection for NAFLD/NASH-associated HCC compared with other HCC etiologies. Demographic, clinicopathological features, and survival outcomes of patients with surgically resected HCC were collected. NAFLD activity score (NAS) and fibrosis score were assessed by focused pathologic review in a subset of patients. Among 492 patients screened, 260 met eligibility (NAFLD/NASH [n = 110], and other etiologies [n = 150]). Median age at diagnosis was higher in the NAFLD/NASH HCC cohort compared with the other etiologies cohort (66.7 vs. 63.4 years, respectively, P = .005), with an increased percentage of female patients (36% vs. 18%, P = .001). NAFLD/NASH-related tumors were more commonly >5 cm (66.0% vs. 45%, P = .001). There were no significant differences in rates of lymphovascular or perineural invasion, histologic grade, or serum AFP levels. The NAFLD/NASH cohort had lower rates of background liver fibrosis, lower AST and ALT levels, and higher platelet counts (P < .01 for all). Median overall survival (OS) was numerically shorter in NAFLD/NASH vs other etiology groups, however, not statistically significant. Patients with NAFLD/NASH-related HCC more commonly lacked liver fibrosis and presented with larger HCCs compared with patients with HCC from other etiologies. No differences were seen in rates of other high-risk features or survival. With the caveat of sample size and retrospective analysis, this supports a similar decision-making approach regarding surgical resection for NAFLD/NASH and other etiology-related HCCs.

Non-alcoholic fatty liver disease-related hepatocellular carcinoma.

Non-alcoholic fatty liver disease (NAFLD), one of the most common causes of liver disease, is an increasingly common cause of hepatocellular carcinoma (HCC). Several demographic, clinical, and genetic factors contribute to HCC risk in NAFLD patients, which may inform risk stratification scores. Proven efficacious approaches to primary prevention approach in patients with non-viral liver disease remain an area of need. Semi-annual surveillance is associated with improved early tumor detection and reduced HCC-related mortality; however, patients with NAFLD have several challenges to effective surveillance, including under-recognition of at-risk patients, low surveillance utilization in clinical practice, and lower sensitivity of current tools for early-stage HCC detection. Treatment decisions are best made in a multidisciplinary fashion and are informed by several factors including tumor burden, liver dysfunction, performance status, and patient preferences. Although patients with NAFLD often have larger tumor burden and increased comorbidities compared to counterparts, they can achieve similar post-treatment survival with careful patient selection. Therefore, surgical therapies continue to provide a curative treatment option for patients diagnosed at an early stage. Although there has been debate about the efficacy of immune checkpoint inhibitors in patients with NAFLD, current data are insufficient to change treatment selection based on liver disease etiology.

HBV-infected hepatocellular carcinoma can be robustly classified into three clinically relevant subgroups by a novel analytical protocol.

Liver cancer is the third leading cause of cancer-related death worldwide, and hepatocellular carcinoma (HCC) accounts for a relatively large proportion of all primary liver malignancies. Among the several known risk factors, hepatitis B virus (HBV) infection is one of the important causes of HCC. In this study, we demonstrated that the HBV-infected HCC patients could be robustly classified into three clinically relevant subgroups, i.e. Cluster1, Cluster2 and Cluster3, based on consistent differentially expressed mRNAs and proteins, which showed better generalization. The proposed three subgroups showed different molecular characteristics, immune microenvironment and prognostic survival characteristics. The Cluster1 subgroup had near-normal levels of metabolism-related proteins, low proliferation activity and good immune infiltration, which were associated with its good liver function, smaller tumor size, good prognosis, low alpha-fetoprotein (AFP) levels and lower clinical stage. In contrast, the Cluster3 subgroup had the lowest levels of metabolism-related proteins, which corresponded with its severe liver dysfunction. Also, high proliferation activity and poor immune microenvironment in Cluster3 subgroup were associated with its poor prognosis, larger tumor size, high AFP levels, high incidence of tumor thrombus and higher clinical stage. The characteristics of the Cluster2 subgroup were between the Cluster1 and Cluster3 groups. In addition, MCM2-7, RFC2-5, MSH2, MSH6, SMC2, SMC4, NCPAG and TOP2A proteins were significantly upregulated in the Cluster3 subgroup. Meanwhile, abnormally high phosphorylation levels of these proteins were associated with high levels of DNA repair, telomere maintenance and proliferative features. Therefore, these proteins could be identified as potential diagnostic and prognostic markers. In general, our research provided a novel analytical protocol and insights for the robust classification, treatment and prevention of HBV-infected HCC.

External validation of the PAGE-B score for HCC risk prediction in people living with HIV/HBV coinfection

HBV coinfection is common among people living with HIV (PLWH) and is the most important cause of hepatocellular carcinoma (HCC). While risk prediction tools for HCC have been validated in patients with HBV monoinfection, they have not been evaluated in PLWH. Thus, we performed an external validation of PAGE-B in people with HIV/HBV coinfection. We included data on PLWH from four European cohorts who were positive for HBsAg and did not have HCC before starting tenofovir. We estimated the predictive performance of PAGE-B for HCC occurrence over 15 years in patients receiving tenofovir-containing antiretroviral therapy. Model discrimination was assessed after multiple imputation using Cox regression with the prognostic index as a covariate, and by calculating Harrell's c-index. Calibration was assessed by comparing our cumulative incidence with the PAGE-B derivation study using Kaplan-Meier curves. In total, 2,963 individuals with HIV/HBV coinfection on tenofovir-containing antiretroviral therapy were included. PAGE-B was <10 in 26.5%, 10-17 in 57.7%, and ≥18 in 15.7% of patients. Within a median follow-up of 9.6 years, HCC occurred in 68 individuals (2.58/1,000 patient-years, 95% CI 2.03-3.27). The regression slope of the prognostic index for developing HCC within 15 years was 0.93 (95% CI 0.61-1.25), and the pooled c-index was 0.77 (range 0.73-0.80), both indicating good model discrimination. The cumulative incidence of HCC was lower in our study compared to the derivation study. A PAGE-B cut-off of <10 had a negative predictive value of 99.4% for the development of HCC within 5 years. Restricting efforts to individuals with a PAGE-B of ≥10 would spare unnecessary HCC screening in 27% of individuals. For individuals with HIV/HBV coinfection, PAGE-B is a valid tool to determine the need for HCC screening. Chronic HBV infection is the most important cause of hepatocellular carcinoma (HCC) among people living with HIV. Valid risk prediction may enable better targeting of HCC screening efforts to high-risk individuals. We aimed to validate PAGE-B, a risk prediction tool that is based on age, sex, and platelets, in 2,963 individuals with HIV/HBV coinfection who received tenofovir-containing antiretroviral therapy. In the present study, PAGE-B showed good discrimination, adequate calibration, and a cut-off of <10 had a negative predictive value of 99.4% for the development of HCC within 5 years. These results indicate that PAGE-B is a simple and valid risk prediction tool to determine the need for HCC screening among people living with HIV and HBV.

THE ROLE OF PIVKAII ONCOMARKER IN HEPATOCELLULAR CARCINOMA:&#x0D; А LITERATURE REVIEW

Relevance: Hepatocellular carcinoma (HCC) ranks sixth among the most common malignant neoplasms in the world and accounts for about&#x0D; 5.6% of all human malignant neoplasms. Despite encouraging progress in the diagnosis and treatment of HCC, the prognosis remains unsatisfactory, i.e., with a 5-year overall survival rate below 10.3%. However, the survival rate can reach 50-74% if early detection and therapeutic intervention are carried out on time. However, unfortunately, about 50% of HCC cases are diagnosed at a late stage.&#x0D; The protein induced by the absence of vitamin K or antagonist-II (PIVKA-II), also known as Dez-γ-carboxyprothrombin (DCP), is another&#x0D; marker specific to HCC. In several studies, elevated PIVKA-II serum levels were associated with HCC. Many authors have proven the PIVKA-II&#x0D; applicability for HCC monitoring.&#x0D; This study aimed to compare the efficiency of alpha-fetoprotein and des-gamma-carboxyprothrombin serological markers in HCC.&#x0D; Methods: The study included the review of published articles on the causes of HCC and the analysis of literature to compare cancer markers&#x0D; efficacy, including PIVKA-II and alpha-fetoprotein (AFP), in detecting HCC.&#x0D; Results: The published results evidence an important role of PIVKA-II in HCC early detection, since PIVKA-II elevation in risk-group patients&#x0D; predicts HCC development in two years. Higher PIVKA-II levels can indicate a bigger tumor or a higher clinical stage. Besides, HCC patients with&#x0D; metastasis to the lymph nodes and distant metastasis had much higher PIVKA-II levels compared to non-metastatic patients. So, high PIVKA-II&#x0D; levels can to a certain extent reflect poor prognosis in HCC patients.&#x0D; Conclusion: The reviewed publications report much higher PIVKA-II serum levels in patients with HCC compared to patients with benign liver&#x0D; diseases or healthy people. Besides, PIVKA-II has a higher diagnostic capacity than AFP due to its higher levels, sensitivity, and specificity. Thus,&#x0D; we can expect high sensitivity and efficiency of the PIVKA-II oncomarker in HCC early diagnostics.

Emerging novel targets for nonalcoholic fatty liver disease treatment: Evidence from recent basic studies.

Nonalcoholic fatty liver disease (NAFLD), a leading chronic disease worldwide, affects approximately a quarter of the global population. Nonalcoholic steatohepatitis (NASH) is an advanced form of NAFLD and is more likely to progress to liver fibrosis than simple steatosis. NASH is also identified as the most rapidly growing cause of hepatocellular carcinoma. Although in the past decade, several phase II/III clinical trials have shown promising results in the use of novel drugs targeting lipid synthase, farnesoid X receptor signaling, peroxisome proliferator-activated receptor signaling, hepatocellular injury, and inflammatory signaling, proven pharmaceutical agents to treat NASH are still lacking. Thus, continuous exploration of the mechanism underlying the pathogenesis of NAFLD and the identification of novel therapeutic targets remain urgent tasks in the field. In the current review, we summarize studies reported in recent years that not only provide new insights into the mechanisms of NAFLD development but also explore the possibility of treating NAFLD by targeting newly identified signaling pathways. We also discuss evidence focusing on the intrahepatic targets involved in the pathogenesis of NAFLD as well as extrahepatic targets affecting liver metabolism and function.

Tumorigenesis from non-alcoholic steatohepatitis to hepatocellular carcinoma.

Non-alcoholic steatohepatitis (NASH) with metabolic syndrome is increasing to be a main cause of hepatocellular carcinoma (HCC). However, the mechanism of tumorigenesis in NASH induced HCC is still not clear. In this perspective, we will discuss the recent progress that has been made to understand the genetic change and the immune microenvironment of HCC, and the remaining questions. Based on the current study, NASH-HCC is likely to have novel mechanism, which needs more investigation in future.

A double-edged sword: The HBV-induced non-coding RNAs alterations in hepatocellular carcinoma

Non-coding RNAs are speculated to exert important regulatory functions at the level of gene expression, oncogenesis, and many other pathologies. Hepatitis B virus (HBV) infection is a leading cause of hepatocellular carcinoma (HCC), and some studies have shown that the expression of non-coding RNAs has an assignable effect on the development of HBV-induced HCC. In this context, the functions and molecular mechanisms of the HBVinduced non-coding RNA expression in the development of hepatoma have attracted increasing attention. This review covers the progress in the exploration of the relationship between HBV-induced hepatoma and non-coding RNA expression, cataloging the recent reports about the roles of non-coding RNAs in HBV-induced hepatoma into five classes, including (1) modulation of metabolism in hepatic cancer, (2) aggravation of inflammation and hepatic fibrosis, (3) alteration of the tumor immune microenvironment, (4) non-coding RNA N6-methyladenosine modification, and a seemingly opposite process, (5) the suppression of the progression of HBV-related HCC. All evidence supports non-coding RNAs as promising novel targets for the early diagnosis and treatments for HCC.

HBV X Protein Induces Degradation of UBXN7, a Novel Negative Regulator of NF-κB Signaling, to Promote HBV Replication.

Chronic hepatitis B virus (HBV) infection is a leading cause of hepatocellular carcinoma. However, the function and mechanism of the effect of HBV on host protein ubiquitination remain largely unknown. We aimed at characterizing whether and how HBV promotes self-replication by affecting host protein ubiquitination. In this study, we identified UBXN7, a novel inhibitor for nuclear factor kappa B (NF-κB) signaling, was degraded via interaction with HBV X protein (HBx) to activate NF-κB signaling and autophagy, thereby affecting HBV replication. The expression of UBXN7 was analyzed by Western blot and quantitative reverse transcription polymerase chain reaction in HBV-transfected hepatoma cells and HBV-infected primary human hepatocytes (PHHs). The effects of UBXN7 on HBV replication were analyzed by using invitro and invivo assays, including stable isotope labeling by amino acids in cell culture (SILAC) analysis. Changes in HBV replication and the associated molecular mechanisms were analyzed in hepatoma cell lines. SILAC analyses showed that the ubiquitination of UBXN7 was significantly increased in HepG2.2.15 cells compared with control cells. After HBV infection, HBx protein interacted with UBXN7 to promote K48-linked ubiquitination of UBXN7 at K99, leading to UBXN7 degradation. On the other hand, UBXN7 interacted with the ULK domain of IκB kinase β through its ubiquitin-associating domain to facilitate its degradation. This in turn reduced NF-κB signaling, leading to reduced autophagy and consequently decreased HBV replication.

THE ROLE OF PIVKAII ONCOMARKER IN HEPATOCELLULAR CARCINOMA: А LITERATURE REVIEW

Relevance: Hepatocellular carcinoma (HCC) ranks sixth among the most common malignant neoplasms in the world and accounts for about 5.6% of all human malignant neoplasms. Despite encouraging progress in the diagnosis and treatment of HCC, the prognosis remains unsatisfactory, i.e., with a 5-year overall survival rate below 10.3%. However, the survival rate can reach 50-74% if early detection and therapeutic intervention are carried out on time. However, unfortunately, about 50% of HCC cases are diagnosed at a late stage.&#x0D; The protein induced by the absence of vitamin K or antagonist-II (PIVKA-II), also known as Dez-γ-carboxyprothrombin (DCP), is another marker specific to HCC. In several studies, elevated PIVKA-II serum levels were associated with HCC. Many authors have proven the PIVKA-II applicability for HCC monitoring.&#x0D; This study aimed to compare the efficiency of alpha-fetoprotein and des-gamma-carboxyprothrombin serological markers in HCC.&#x0D; Methods: The study included reviewing published articles on the causes of HCC and analyzing literature to compare cancer markers’ efficacy, including PIVKA-II and alpha-fetoprotein (AFP), in detecting HCC.&#x0D; Results: The published results evidence an important role of PIVKA-II in HCC early detection since PIVKA-II elevation in risk-group patients predicts HCC development in two years. Higher PIVKA-II levels can indicate a bigger tumor or a higher clinical stage. Besides, HCC patients with metastasis to the lymph nodes and distant metastasis had much higher PIVKA-II levels than non-metastatic patients. So, high PIVKA-II levels can, to a certain extent, reflect poor prognosis in HCC patients.&#x0D; Conclusion: The reviewed publications report much higher PIVKA-II serum levels in patients with HCC than in patients with benign liver diseases or healthy people. Besides, PIVKA-II has a higher diagnostic capacity than AFP due to its higher levels, sensitivity, and specificity. Thus, we can expect high sensitivity and efficiency of the PIVKA-II tumor marker in early HCC diagnostics.

Evaluation of Amino Acids Profile as Non-Invasive Biomarkers of Hepatocellular Carcinoma in Egyptians.

Hepatocellular carcinoma (HCC) is the most dangerous complication of chronic liver disease. It is a multifactorial complicated disease. Hepatitis C and hepatitis B viruses (HCV and HBV, respectively) represent the main causes of HCC in Egypt. Early diagnosis is very important to aid in early intervention. The goal of this research is to evaluate the metabolic role of different amino acids as non-invasive biomarkers over the course of HCC. This study included 302 participants with 97 diagnosed, untreated HCC patients, 81 chronic HCV patients, 56 chronic HBV patients, 18 co-infected patients, and a control group of 50 normal age and gender-matched individuals. All participants provided complete medical histories and underwent complete clinical examinations, abdominal ultrasonography and/or computed tomography, routine laboratory investigations, estimation of serum α-fetoprotein, and determination of amino acid levels using ultra-performance liquid chromatography (UPLC MS/MS). This work revealed a decline in branched chain amino acids (BCAA) and increase in aromatic amino acids (AAA) among infected groups (HCC, HBV, HCV, and co-infected patients) compared to control subjects and a marked change in Fisher's and the BCAAs/tyrosine molar concentration ratios (BTR) between controls and infected groups. Different amino acids could be used as non-invasive markers to discriminate and follow chronic hepatitis patients to predict the course of HCC.

Characteristics and analysis of the costs of drug provision of patients with non-alcoholic fatty liver disease in Moscow

Introduction. The change in the classical etiological paradigm of hepatocellular carcinoma has led to the fact that non-alcoholic fatty liver disease is now the fastest growing cause of hepatocellular carcinoma in the world. Purpose. To study the range of medicines and analyze the costs of drug provision for a determined population of patients with non-alcoholic fatty liver disease in Moscow over a three-year period. Materials and methods. Based on data from the Moscow Department of Health on preferential drug coverage for the period from 2019 to 2021 the range of drug therapy and the economic costs. Results. For 3 years, seventeen thousand two hundred ten patients with a diagnosis of non-alcoholic fatty liver disease received preferential drug coverage in Moscow, and the amount of costs minus payment by the patient amounted to 49,102,963.97 rubles. The most prescribed drugs were drugs from the group of “hepatoprotectors”. An increase in the population of patients with non-alcoholic fatty liver disease over a three-year period by 1.7 times, an increase in the cost of its drug provision. Along with this, the prevalence in the spectrum of drugs was revealed: glycyrrhizic acid + phospholipids; phospholipids; ursodeoxycholic acid; ademetionine is not found in international practice and is not funded under the nosology “non-alcoholic fatty liver disease”. Limitations. When studying options for drug provision of patients with non -alcoholic fatty liver disease, we had an assessment of 17,210 observations, which is a sufficient reference sample Conclusion. Analysis of the qualitative and quantitative composition of the population of patients with non-alcoholic fatty liver disease can be an effective tool for predicting the social and economic burden of the disease and can be used for rational planning of budget expenditures of all levels.

Quantification of DNA methylation for carcinogenic risk estimation in patients with non-alcoholic steatohepatitis

BackgroundIn recent years, non-alcoholic steatohepatitis (NASH) has become the main cause of hepatocellular carcinoma (HCC). As a means of improving the treatment of NASH-related HCCs based on early detection, this study investigated the feasibility of carcinogenic risk estimation in patients with NASH.ResultsNormal liver tissue (NLT), non-cancerous liver tissue showing histological findings compatible with non-alcoholic fatty liver from patients without HCC (NAFL-O), non-cancerous liver tissue showing NASH from patients without HCC (NASH-O), non-cancerous liver tissue showing non-alcoholic fatty liver from patients with HCC (NAFL-W), non-cancerous liver tissue showing NASH from patients with HCC (NASH-W) and NASH-related HCC were analyzed. An initial cohort of 171 tissue samples and a validation cohort of 55 tissue samples were used. Genome-wide DNA methylation screening using the Infinium HumanMethylation450 BeadChip and DNA methylation quantification using high-performance liquid chromatography (HPLC) with a newly developed anion-exchange column were performed. Based on the Infinium assay, 4050 CpG sites showed alterations of DNA methylation in NASH-W samples relative to NLT samples. Such alterations at the precancerous NASH stage were inherited by or strengthened in HCC samples. Receiver operating characteristic curve analysis identified 415 CpG sites discriminating NASH-W from NLT samples with area under the curve values of more than 0.95. Among them, we focused on 21 CpG sites showing more than 85% specificity, even for discrimination of NASH-W from NASH-O samples. The DNA methylation status of these 21 CpG sites was able to predict the coincidence of HCC independently from histopathological findings such as ballooning and fibrosis stage. The methylation status of 5 candidate marker CpG sites was assessed using a HPLC-based system, and for 3 of them sufficient sensitivity and specificity were successfully validated in the validation cohort. By combining these 3 CpG sites including the ZC3H3 gene, NAFL-W and NASH-W samples from which HCCs had already arisen were confirmed to show carcinogenic risk with 95% sensitivity in the validation cohort.ConclusionsAfter a further prospective validation study using a larger cohort, carcinogenic risk estimation in liver biopsy specimens of patients with NASH may become clinically applicable using this HPLC-based system for quantification of DNA methylation.

Immunohistochemical expression of CB1 receptors in the liver of patients with HBV related-HCC

Background and aimsThe most common cause of hepatocellular carcinoma (HCC), which has a high death rate, is hepatitis B virus (HBV) infection. This study aimed to examine immunoexpression of CB1 receptors in the livers of patients with HBV-related HCC in comparison with HCC and chronic HBV as well as healthy people.Patients and methodsParticipants in this case-controlled study were patients with only HBV (HBV; 40), only HCC (HCC; 41), and HBV-related HCC (HBV + HCC; 40) and a healthy control group (C; 30). Tissue expression of CB1 at the protein level was studied using immunohistochemical methods. ResultsAll groups were significantly different in terms of expression of CB1 protein (P < 0.001). The expression levels of CB1 in the liver tissue of the HBV and C groups were not significantly different (P = 0.072). The expression levels of CB1 in the liver tissue of the HBV-related HCC and HCC groups had a statistically significant increase compared to the C and HBV groups (P < 0.001). Also, the CB1 expression levels in the liver tissues of HBV-related HCC and HCC groups were significantly different (P = 0.008). The sensitivity and specificity of immunohistochemistry tests in the diagnosis of HCC using CB1 were 63.4 and 91.2, respectively. Positive and negative predictive values were 90.0 % and 65.1 %, respectively. There was no relationship between the expression of CB1and other clinicopathological variables (P < 0.05). ConclusionThe present findings revealed a tumor promoting function of the CB1 receptor in HCC. CB1 is also a pathological valuable factor for identifying the pathway of inflammation during infection.

Profile of Hepatocellular Carcinoma: An Experience from a Tertiary Care Hospital

Background and Aim: Hepatocellular carcinoma (HCC) is the primary liver cancer around the world, and it has now become the third leading cause of cancer-related mortality. With an increasing global incidence, HCC is becoming a major health burden. The incidence of HCC varies from 0.3% to 1.6% in Asian countries. The present study aimed to assess the clinical and etiological profile of hepatocellular carcinoma in a tertiary care hospital. Methodology: This retrospective study was conducted on 124 hepatocellular carcinoma patients in the Department of Gastroenterology, Jinnah Hospital, Lahore from June 2019 to May 2022. All the patients of age 10 years to 75 years fulfilling the diagnostic criteria were enrolled in this study. Patient’s cytohistological, clinical, radiological, and etiological data were recorded and analyzed. Individual relevant features were noted in pre-designed proforma. SPSS version 25 was used for data analysis. Results: Of the total 124 HCC patients, there were 78 (62.9%) male and 46 (37.1%) females. The overall mean age was 48.63±6.78 years. Liver cirrhosis was present in 91 (73.4%) patients out of which 14 (15.4%) were asymptomatic. The prevalent signs of hepatic decompensation were ascites 67 (54%) and Jaundice 23 (18.5%). Based on etiological data, the prevalence of the cryptogenic, Hepatitis B, and C were 58 (46.8%), 20 (16.1%), and 6 (4.8%) respectively. Tumor thrombosis was found in 18 (14.5%). Based on biopsy specimens, HCC with steatosis were seen in 19 (15.3%) patients. During surveillance, about 32 (25.8%) cirrhotic HCC patients were diagnosed. Conclusion: The present study found that the most prevalent etiology was hepatocellular carcinoma. Most patients were asymptomatic and had major complaints of abdominal pain with certain specific comorbidities. Hypertension, diabetes, and ischemic heart disease were the specific co-morbidities in hepatocellular carcinoma patients. In our population, the prevalent cause of HCC was Hepatitis B. Keywords: Clinical profile, Etiology, Hepatocellular Carcinoma

Ectopic clotting factor VIII expression and misfolding in hepatocytes as a cause for hepatocellular carcinoma

Hemophilia A gene therapy targets hepatocytes to express B domain deleted (BDD) clotting factor VIII (FVIII) to permit viral encapsidation. Since BDD is prone to misfolding in the endoplasmic reticulum (ER) and ER protein misfolding in hepatocytes followed by high-fat diet (HFD) can cause hepatocellular carcinoma (HCC), we studied how FVIII misfolding impacts HCC development using hepatocyte DNA delivery to express three proteins from the same parental vector: (1) well-folded cytosolic dihydrofolate reductase (DHFR); (2) BDD-FVIII, which is prone to misfolding in the ER; and (3) N6-FVIII, which folds more efficiently than BDD-FVIII. One week after DNA delivery, when FVIII expression was undetectable, mice were fed HFD for 65weeks. Remarkably, all mice that received BDD-FVIII vector developed liver tumors, whereas only 58% of mice that received N6 and no mice that received DHFR vector developed liver tumors, suggesting that the degree of protein misfolding in the ER increases predisposition to HCC in the context of an HFD and in the absence of viral transduction. Our findings raise concerns of ectopic BDD-FVIII expression in hepatocytes in the clinic, which poses risks independent of viral vector integration. Limited expression per hepatocyte and/or use of proteins that avoid misfolding may enhance safety.

Hepatocyte phosphatase DUSP22 mitigates NASH-HCC progression by targeting FAK

Nonalcoholic steatohepatitis (NASH), a common clinical disease, is becoming a leading cause of hepatocellular carcinoma (HCC). Dual specificity phosphatase 22 (DUSP22, also known as JKAP or JSP-1) expressed in numerous tissues plays essential biological functions in immune responses and tumor growth. However, the effects of DUSP22 on NASH still remain unknown. Here, we find a significant decrease of DUSP22 expression in human and murine fatty liver, which is mediated by reactive oxygen species (ROS) generation. Hepatic-specific DUSP22 deletion particularly exacerbates lipid deposition, inflammatory response and fibrosis in liver, facilitating NASH and non-alcoholic fatty liver disease (NAFLD)-associated HCC progression. In contrast, transgenic over-expression, lentivirus or adeno-associated virus (AAV)-mediated DUSP22 gene therapy substantially inhibit NASH-related phenotypes and HCC development in mice. We provide mechanistic evidence that DUSP22 directly interacts with focal adhesion kinase (FAK) and restrains its phosphorylation at Tyr397 (Y397) and Y576 + Y577 residues, subsequently prohibiting downstream activation of extracellular signal-regulated kinase 1/2 (ERK1/2) and nuclear factor-κB (NF-κB) cascades. The binding of DUSP22 to FAK and the dephosphorylation of FAK are indispensable for DUSP22-meliorated NASH progression. Collectively, our findings identify DUSP22 as a key suppressor of NASH-HCC, and underscore the DUSP22-FAK axis as a promising therapeutic target for treatment of the disease.