Search Results (4,086)

Chronic hepatitis B virus (CHB) infection remains a major cause of hepatocellular carcinoma (HCC), yet the premalignant microenvironment that links to HBV-associated HCC (HBV-HCC) is still poorly defined. This review synthesizes evidence that HBV-infected hepatocytes function as signaling hubs that, through microRNA (miRNA)-regulated crosstalk with Kupffer cells, liver sinusoidal endothelial cells, hepatic stellate cells and cancer-associated fibroblasts (CAFs), progressively remodel the liver from an antiviral tissue into a premalignant and early tumor microenvironment. Across the HBV-HCC continuum, a core set of dysregulated miRNAs, including miR-21, miR-29a/b, miR-122, miR-146a, miR-155, miR-200a, miR-126, miR-210 and the miR-130/301 family, coordinates transition from innate antiviral responses to HSC activation, extracellular matrix deposition, mechanotransduction, angiogenesis, chronic inflammation and cancer-associated CAF programing. By mapping these stage-specific miRNA networks onto acute infection, CHB, early fibrogenesis, advanced fibrosis and CAF-rich dysplastic states, the review reframes HBV-HCC pathogenesis as a sequence of miRNA-guided hepatocyte–stromal states rather than a purely hepatocyte-intrinsic process. This perspective suggests that composite, cell-type-resolved miRNA signatures in serum or liver tissue could serve as biomarkers for identifying CHB patients who are entering a premalignant microenvironment before conventional surveillance markers become abnormal. It further highlights miRNA hubs that couple antiviral, fibrogenic, angiogenic and CAF-associated signaling as potential therapeutic targets for reprograming the HBV-driven premalignant microenvironment, with the long-term goal of intercepting HBV-HCC development at earlier, microenvironmentally defined stages. Full article

Aflatoxin B1 (AFB1) is a potent foodborne mycotoxin classified as a Group 1 human carcinogen by the International Agency for Research on Cancer (IARC) and is widely recognized for its causal role in hepatocellular carcinoma (HCC). Beyond its well-established hepatotoxicity, increasing evidence indicates that AFB1 also disrupts intestinal homeostasis by impairing epithelial barrier integrity, reducing mucus production, altering gut microbial composition, and disturbing microbial metabolism. These changes promote increased intestinal permeability, facilitating the translocation of bacteria and microbial products and contributing to chronic inflammation through the gut–liver axis. Recent experimental studies further suggest that alterations in short-chain fatty acid (SCFA) production and microbiota-dependent signaling pathways may actively mediate AFB1-induced intestinal and hepatic injury. Although direct evidence linking AFB1-induced dysbiosis to extrahepatic carcinogenesis remains limited, growing evidence indicates that persistent barrier dysfunction, microbial imbalance, and chronic inflammation may create a microenvironment favorable for tumor initiation and progression in tissues beyond the liver. This review critically summarizes current evidence regarding the effects of AFB1 on intestinal barrier function, gut microbiota dysbiosis, bacterial translocation, microbial metabolites, and the gut–liver axis while evaluating the mechanistic evidence supporting these interactions and highlighting the major knowledge gaps that should be addressed in future research. Full article

Geranylgeranoic acid (GGA) is an endogenous acyclic diterpenoid metabolite of the mevalonate pathway that has been implicated in programmed cell death in hepatoma cells and may contribute to the elimination of premalignant hepatocytes. Recent metabolomic studies have identified 2,3-dihydrogeranylgeranoic acid (2,3-diGGA), an α-saturated derivative of GGA with reduced cell-death-inducing activity compared with GGA. This concise review examines the hypothesis that diversion of bioactive GGA toward 2,3-diGGA formation represents a metabolic shift that may attenuate hepatic tumor surveillance and influence hepatocellular carcinoma susceptibility. The review summarizes current evidence for endogenous GGA biosynthesis through the mevalonate pathway, MAO-B- and CYP3A4-associated GGA formation, and the tissue- and age-associated distribution of GGA and 2,3-diGGA. It also discusses the proposed 2,3-diGGA-forming activity, whose molecular identity remains unknown, and evaluates the potential utility of the GGA/2,3-diGGA ratio as a candidate biomarker. Finally, future experimental strategies are outlined to identify the responsible enzyme, clarify causality in cellular and animal models, and validate this metabolic framework in human liver tissues, chronic liver disease cohorts, and HCC-associated settings. Full article

Hepatocellular carcinoma (HCC) remains a major global health challenge and one of the leading causes of cancer-related mortality, with advanced disease continuing to be associated with limited therapeutic options and substantial heterogeneity in response to systemic treatment. Recent evidence has established the gut microbiota, through the gut–liver axis, as a critical determinant of immunotherapy efficacy, while also influencing antitumor immunity and liver carcinogenesis. Microbial dysbiosis may promote chronic inflammation, intestinal barrier disruption, bacterial translocation, and immune dysfunction, thereby contributing to hepatocarcinogenesis. Moreover, gut microbial composition and microbial-derived metabolites, including bile acids, short-chain fatty acids (SCFAs), and inosine, have been associated with modulation of antitumor immune responses and differential outcomes to immune checkpoint inhibitors (ICIs). Emerging clinical evidence in HCC has identified distinct gut microbial signatures associated with response to nivolumab, pembrolizumab, and atezolizumab-based regimens, including enrichment of Akkermansia muciniphila and SCFA-producing taxa such as Ruminococcaceae, Roseburia, and Prevotella in responders. However, these findings remain inconsistent across studies, with no reproducible microbial signature identified because of small cohort sizes, heterogeneous patient populations, geographic variation, cirrhosis-related confounding factors, and methodological differences in microbiome analysis. This review summarizes the current understanding of microbiome–immune interactions in HCC, examines mechanistic pathways linking the microbiota to immunotherapy response, critically evaluates available clinical evidence, and discusses current limitations and future therapeutic strategies, including fecal microbiota transplantation, probiotics, dietary modulation, and engineered bacterial platforms. Collectively, microbiome-based approaches may contribute to the development of personalized immunotherapeutic strategies in HCC, although larger standardized prospective studies are required before microbiome-derived biomarkers can be implemented in routine clinical practice. Full article

Hepatocellular carcinoma (HCC) is one of the malignant tumors with high incidence and mortality rates worldwide. Given the poor prognosis of patients with HCC, it is crucial to explore the molecular mechanisms underlying HCC development and to evaluate prognostic markers. Differential expression analysis followed by univariate Cox, LASSO, and multivariate Cox regression identified four genes (EPO, SOCS2, IL18RAP, and KPNA2), and a Cox-based risk score was evaluated in the TCGA-LIHC cohort and externally in GSE14520 using Kaplan–Meier and time-dependent ROC analyses. Bulk, single-cell, and protein resources provided convergent expression context. Survival machine-learning analysis using observed overall-survival time and censoring status identified Cox–Ridge as the best-performing model in TCGA-LIHC, with more modest performance in GSE14520, and immune profiling revealed risk-group-associated differences in estimated immune and stromal components, immune-cell composition, and immune-checkpoint expression. The oncoPredict/GDSC2 screen highlighted five potential drug candidates for experimental prioritization. Because the drug screen is based on computationally predicted sensitivities, these findings should be regarded as hypothesis-generating and require validation in prospective cohorts and experimental systems before clinical translation. Full article

Microvascular invasion (MVI) is a critical prognostic indicator for recurrence and survival in hepatocellular carcinoma (HCC); however, its accurate preoperative assessment remains clinically challenging. Postoperative histopathology is subject to sampling bias and time delays, while traditional imaging techniques lack the molecular specificity required to predict MVI. Liquid biopsy, through the analysis of circulating tumor DNA (ctDNA), circulating tumor cells (CTCs), circulating tumor RNA (ctRNA), and extracellular vesicles (EVs), provides a minimally invasive approach for capturing tumor-derived molecular and cellular signals associated with vascular invasion. This narrative review comprehensively summarizes the current evidence linking these four liquid biopsy analyte categories to MVI in HCC, evaluates their integration into multi-omics predictive models, including multi-marker, clinicopathological-integrated, and imaging-integrated strategies, and proposes an evidence-level framework that categorizes blood biomarkers according to the strength of their support for MVI prediction, distinguishing direct histopathological validation from indirect associations with aggressive tumor biology. Key challenges are critically examined, including the variable specificity of individual biomarkers for MVI, the lack of head-to-head comparative studies, the absence of standardized pre-analytical and analytical protocols, and the methodological limitations of current prediction models. As a narrative review, this work does not employ systematic review methodology, and the evidence synthesis should be interpreted accordingly. The review provides a framework for understanding how liquid biopsy-based MVI risk stratification may inform surgical and perioperative decision-making following prospective validation. Full article

Hepatocellular carcinoma (HCC) is one of the most prevalent cancers worldwide and exhibits considerable biological heterogeneity in both molecular and clinical characteristics. The diverse molecular alterations and clinical manifestations of HCC indicate substantial heterogeneity across patient subgroups. This study aimed to identify novel therapeutic targets and predictive biomarkers associated with HCC using an integrative bioinformatics approach. High-throughput genomic datasets were obtained from the UCSC Xena browser to retrieve mRNA HTSeq-count data from the TCGA-HCC cohort. Gene co-expression network (GCN), protein–protein interaction network (PPIN), and enrichment analyses were performed to identify key dysregulated genes and their biological significance. Integrated network analyses identified three dysregulated hub genes, namely CXCR2, TLR2, and TLR4. Genomic alterations in these genes were further evaluated across tumor samples in the TCGA-HCC cohort. Kaplan–Meier (KM) survival analysis demonstrated that lower CXCR2 mRNA expression was significantly associated with poorer overall survival (OS) and recurrence-free survival (RFS). Furthermore, TIMER and UALCAN analyses revealed significant associations between CXCR2 expression and tumor purity, as well as immune cell infiltration levels, including T cells, macrophages, dendritic cells (DCs), and neutrophils. These findings suggest that CXCR2 is significantly associated with the immune microenvironment of HCC and represents a potential prognostic biomarker whose biological role warrants further mechanistic investigation. Full article

Background: Predicting recurrence after liver transplantation (LT) for hepatocellular carcinoma (HCC) remains important for post-transplant risk stratification and surveillance planning. The Milan criteria discriminate only moderately and some machine-learning re-analyses report overly optimistic results because of information leakage. Aim: The current study aimed to re-evaluate a previously published transplant cohort under a leakage-free survival-analysis framework and to benchmark post-transplant, explant-informed survival learners against the Milan criteria as a fixed pre-transplant reference. We hypothesised moderate rather than near-perfect discrimination, similar performance across learners of differing complexity, and better discrimination than the Milan criteria. Methods: This secondary analysis included 356 patients with HCC who underwent LT. The primary endpoint was recurrence-free survival, analysed from the observed event indicator and follow-up time rather than from a derived risk label. Seven survival learners were benchmarked with repeated nested cross-validation, using three repeats of a five-fold outer loop with a three-fold inner tuning loop. All data-dependent preprocessing, including robust multivariable outlier handling and imputation, was fitted within training folds only. Performance was assessed by the concordance indices of Harrell and Uno, the time-dependent area under the curve, the integrated Brier score, calibration, decision-curve analysis and descriptive competing-risk assessment. Results: Recurrence developed in 183 of the 356 patients over a median follow-up of 52 months. Discrimination was moderate rather than near-perfect and similar across learners; the random survival forest ranked highest and the Elastic-Net Cox model performed comparably. All learners showed higher descriptive concordance than the Milan criteria, and dependency-corrected comparisons supported higher concordance for the full-feature Cox model than for the Milan criteria, whereas the random survival forest and Cox did not differ materially. Out-of-fold calibration of the Elastic-Net Cox model at 36 months was acceptable, decision-curve analysis indicated positive net benefit across clinically relevant thresholds, and tumour size and alpha-fetoprotein were the leading contributors to prediction. Findings were stable in ablation and threshold-sensitivity analyses. Conclusions: Leakage-free survival modelling gave moderate but internally validated prediction of post-transplant recurrence and higher concordance than the Milan criteria in this cohort, supporting the stated hypotheses. Careful study design may matter more than architectural complexity in this setting, and leakage-free survival analysis is a practical standard for prognostic modelling in transplant oncology. Full article

Background: The sensitivity of ultrasound plus alpha-fetoprotein (AFP) for early-stage hepatocellular carcinoma (HCC) detection remains suboptimal, prompting interest in blood-based biomarkers as an alternative strategy. The ASAP [age, sex, AFP, PIVKA-II] score has promising performance, although validation in contemporary Western populations is needed. Methods: We conducted a case–control study in which the cases were patients with HCC and the controls had cirrhosis without HCC. AFP, PIVKA-II, LG2m, and GP73 were measured using the Abbott Alinity platform, and ASAP scores were calculated. Sensitivity and specificity for early-stage HCC (BCLC stage 0/A) were compared between ASAP and ultrasound plus AFP using McNemar’s chi-square test. Results: Among 294 patients (median age of 61 years, 67.7% men), 50.7% had HCC (69.1% BCLC 0/A), and 49.3% had cirrhosis. The performance of individual biomarkers for early-stage HCC was moderate to poor (AUROC: AFP 0.71, PIVKA-II 0.73, LG2m 0.51, and GP73 0.53). ASAP, at a cutoff of ≥0.526, demonstrated a sensitivity of 88.3% for early-stage HCC, with specificity of 54.5%. Sensitivity was higher in men and viremic HCV, whereas specificity was higher in younger individuals, women, and Child–Pugh A cirrhosis. In subgroup analysis (n = 178), ASAP had comparable sensitivity to ultrasound plus AFP (85.5% vs. 90.3%; p = 0.51) but lower specificity (53.4% vs. 93.1%; p < 0.001). Using the Youden-optimized cutoff (≥0.804) improved specificity to 83.4% while maintaining a sensitivity of 68.0% for early-stage HCC. Conclusions: ASAP demonstrated high sensitivity for early-stage HCC detection but low specificity. These findings suggest a need for optimized cutoffs prior to implementation in clinical practice. Full article

Hepatocellular carcinoma (HCC) is the most common primary liver cancer and is frequently diagnosed at advanced stages, thereby limiting curative treatment options. Although interventions with curative intent, such as liver resection and ablation, are available, recurrence rates exceed 70%. Recent developments in HCC management have shifted the focus from conventional surgery and targeted therapies to immunotherapy. This review synthesizes current evidence on immunotherapy for curative purposes in HCC, including both neoadjuvant and adjuvant strategies. It evaluates completed and ongoing clinical trials of immune checkpoint inhibitors (ICIs), administered as monotherapy or in combination with anti-angiogenic agents or dual-checkpoint blockade. ICIs may transform the initial treatment paradigm for advanced HCC, although their applications remain in the investigational stage. Preliminary results from the IMbrave050 trial suggested that adjuvant immunotherapy following radical liver resection improves recurrence-free survival (RFS) in high-risk patients. However, the initially promising outcomes of the combination of atezolizumab and bevacizumab compared with active surveillance were not sustained in the median follow-up, and long-term survival results are awaited. Although several studies have demonstrated encouraging preliminary results, a clear survival benefit has not yet been established. Further studies are necessary to establish definitive conclusions. Immunotherapy has a pathophysiological basis and preliminary evidence to redefine HCC management across disease stages; its integration into pre- and post-operative strategies may increase resectability, reduce recurrence, and improve survival. Ongoing trials are expected to clarify its exact role. Full article

Background/Objectives: To investigate early treatment outcomes, including tumor response, predictors of response, and safety of nivolumab plus ipilimumab (Nivo/Ipi) therapy for unresectable hepatocellular carcinoma (uHCC) in a multicenter setting. Methods: We conducted a retrospective analysis of 64 Japanese patients with uHCC who received Nivo/Ipi therapy at participating centers between July 2025 and April 2026. Treatment efficacy was assessed according to RECIST version 1.1, and adverse events (AEs) were evaluated using CTCAE version 5.0. Results: Among the 64 patients, 28 received first-line therapy, and 36 received later-line therapy. The median observation period was 5.4 months. The overall response rate (ORR) was significantly higher in the first-line group (60.7%) than in the later-line group (30.6%) (p = 0.016). ORRs according to CRAFITY score were 25.0% for score 0, 55.6% for score 1, and 53.8% for score 2. The ORR in patients with CRAFITY scores ≥ 1 was significantly higher than in patients with a CRAFITY score of 0 (55.0% vs. 25.0%; p = 0.022). Multivariable analysis of pretreatment variables predicting response revealed first-line treatment (OR 3.83, 95% CI 1.270–11.558; p = 0.017) and a CRAFITY score ≥ 1 (OR 4.02, 95% CI 1.232–13.106; p = 0.021) as significant independent factors. Immune-mediated AEs occurred in 70.3% of patients overall and were grade ≥ 3 in 35.9%. Conclusions: Nivo/Ipi yielded a favorable response when used as first-line therapy. The baseline CRAFITY score may be a useful predictor of response to Nivo/Ipi therapy. Full article

The liver is an immunologically distinctive organ. Portal blood continuously delivers gut-derived antigens and microbial products, requiring hepatic immunity to balance surveillance with restraint. In hepatocellular carcinoma (HCC), this physiology is commonly overlaid by chronic injury, inflammation, and fibrosis, so the background liver forms part of the disease context in which the tumor microenvironment (TME) develops. This review synthesizes how cellular architecture, tumor-intrinsic programs, and structural, metabolic, and microbial conditions interact to shape immune evasion and heterogeneity. HCC provides the principal evidence base, with intrahepatic cholangiocarcinoma (iCCA) used as a structured, biologically distinct comparator. We organize therapies by the microenvironmental barriers they are intended to modify and distinguish established clinical efficacy from evidence that the proposed mechanisms mediate treatment benefit. Single-cell and spatial studies have resolved cellular states and spatial arrangements, including onco-fetal endothelial–myeloid neighborhoods and a macrophage–fibroblast boundary band separating lymphocyte-rich stroma from malignant tissue. These patterns operate within fibrotic and metabolically altered tissue and vary by etiology, spatial context, and tumor type. Vascular endothelial growth factor blockade with immune checkpoint inhibition and dual checkpoint blockade are established first-line options in advanced HCC. Chemo-immunotherapy is established in biliary tract cancer, and IDH1 inhibition has established efficacy in IDH1-mutant cholangiocarcinoma. Myeloid- and stroma-directed strategies, natural-product approaches, and engineered-cell therapies remain preclinical or early clinical. None of the pivotal trials tested whether the proposed microenvironmental mechanism mediated treatment benefit. The liver TME informs treatment selection without yet determining it. Full article

A coordinated extracellular matrix (ECM) and cytoskeletal remodeling are critical drivers of hepatocellular carcinoma (HCC) progression, yet the upstream post-transcriptional mechanisms regulating these processes in spontaneous canine HCC remain poorly defined. Previously, we reported significantly downregulated miRNAs and enriched pathways from differentially expressed genes (DEGs) in canine HCC tissues. In the present study, we investigated putative post-transcriptional target interactions between the prioritized miRNAs and enriched ECM–cytoskeleton pathway-associated genes (n = 34). Integrative bioinformatic analysis prioritized ACTG1 and SDC1 as the key putative targets of cfa-miR-10a based on their concordant prediction by four target-prediction algorithms, high transcript abundance, and inverse correlations with cfa-miR-10a-5p. RNAhybrid and miRanda analyses supported favorable predicted interactions between cfa-miR-10a and the 3′UTRs of both transcripts, with RNAhybrid minimum free energy values ≤ −20 kcal/mol and miRanda scores ≥ 140. In the matched canine transcriptomic subset, cfa-miR-10a and its predicted targets showed inverse expression trends. Cross-species analysis using TCGA-LIHC datasets further demonstrated concordant dysregulation of ACTG1, SDC1 and hsa-miR-10a-5p in human HCC, despite their weak or non-concordant correlation with hsa-miR-10a-5p. Collectively, these exploratory findings identify a candidate cfa-miR-10a–ACTG1/SDC1 network associated with coordinated ECM and cytoskeletal transcriptomic alterations in canine HCC. While cross-species analysis supports the conservation of target gene dysregulation, the divergent miRNA-mRNA correlation patterns highlight potential species-specific post-transcriptional co-expression landscapes that warrant future functional validation. Full article

(1) Background: The Hepatitis B virus (HBV) is characterized by extensive genetic diversity, including low-frequency variants that contribute to disease progression. We aimed to characterize low-frequency variants and evaluate their potential clinical impact. (2) Methods: We utilized 104 HBV near-full-length sequences generated using next-generation sequencing (NGS) from people living with HIV (PLHIV). We used an in-house bioinformatics suite (HBVgenomeR v5.9.7) to filter for low-frequency variants (5–50%), which were compared to escape and drug resistance mutations (DRMs) and hepatocellular carcinoma (HCC)-associated mutations reported at the consensus level. Unclassified variants were characterized by HBV open reading frames (ORFs) to determine mutation frequency per genomic region. (3) Results: A total of six escape mutations were detected in 8/104 (7.7%) sequences, with _surfaceN131T being the most prevalent (5/8). We also observed six DRMs in 30/104 (28.8%), with _rtV173L being the most prevalent (21/30). Truncation mutations were also observed with _rtA181T/_sW172* and _rtM204I/_sW196L being the most prevalent. A total of 8/104 (7.7%) sequences had four variants associated with HCC. The _xP46S was the highest observed HCC-associated mutation at 5/8. We report 1152 unique uncharacterized variants across all ORFs, and these were found in 94/104 (90.4%) sequences. The RNaseH domain had the highest burden (330/1152, 28.6%). (4) Conclusions: Deep sequencing results identified clinically significant mutations, including those below the 20% detection limit of traditional sequencing, that would go unreported. This highlights the possible underreporting of mutational burden in people living with HBV/HIV, indicating the importance of deep sequencing to aid in HBV/HIV understanding and management. Full article

Background/Objectives: Advanced-stage hepatocellular carcinoma is characterized by a very poor prognosis; thus, highly effective medication is still needed. Often overexpressed heat shock protein 90 is a promising target due to its pivotal role in carcinogenesis. Methods: Antiproliferative effects of 246TMP-3SF5 on HepG2 and HuH-7 cells were assessed by crystal violet staining. Apoptosis was evaluated via subG1 peak, caspase-3 activity and PARP cleavage, and ferroptosis via ROS, glutathione and malondialdehyde levels. Migration was assessed by scratch assay, and in ovo models were used to study angiogenesis and drug effects on microtumors. CAM vascular networks were quantified by semi-automatic segmentation, morphometric and box-counting fractal analysis. Molecular docking and molecular dynamics simulation of heat shock protein 90 were carried out using Autodock Vina and Gromacs respectively. Results: Profound dose- and time-dependent antiproliferative effects of 246TMP-3SF5 against HCC cell lines were observed, revealing low micromolecular IC₅₀ values and selectivity for carcinoma cells with selectivity indices > 1. A significant increase in the sub-G1 peak, key effector caspase-3 activity, as well as cleavage of PARP strongly suggested apoptosis playing a crucial role in the antiproliferative effects. Additionally, HuH-7 cells revealed an elevation of reactive oxygen species and both cell lines showed significant glutathione depletion concomitant with an increase in malondialdehyde concentration upon treatment. The observed effect could be partially reversed by applying ferrostatin-1, suggesting ferroptosis as an additional relevant mode of action. Changes in the cell cycle as well as impaired tumor cell migration were observed. Upon treatment, angiogenesis was impaired and mass of microtumors was significantly reduced. Quantitative CAM analysis showed that vascularized area fraction increased in controls but decreased under both 17-AAG and 246TMP-3SF5. Likewise global mean vessel width decreased relative to controls, while box-counting dimension was reduced under 246TMP-3SF5. Molecular docking and molecular dynamics simulation analysis predicts 246TMP-3SF5 to be binding in catalytic site of heat shock protein 90. Conclusions: 246TMP-3SF5 is a promising novel inhibitor meriting further research as a potential treatment against hepatocellular carcinoma. Full article