Hepatocellular carcinoma (HCC) is a biologically and clinically heterogeneous cancer in which the spatial organization of the tumor microenvironment dictates progression, immune escape, and therapeutic response. Traditio...Hepatocellular carcinoma (HCC) is a biologically and clinically heterogeneous cancer in which the spatial organization of the tumor microenvironment dictates progression, immune escape, and therapeutic response. Traditional genomic and transcriptomic profiling have revealed key molecular pathways, yet they do not capture the spatial relationships that govern tumor-immune-stromal interactions. Spatial omics technologies now bridge this gap, allowing molecular profiling within intact tissue architecture and revealing how cellular neighborhoods, immune barriers, and stromal niches shape tumor behavior. In this review, we summarize recent advances in technologies like spatial transcriptomics, proteomics, genomics, epigenomics, and metabolomics as applied to HCC. We discuss how these technologies have illuminated the spatial dynamics of the invasive margin, macrophage and fibroblast niches, and immune evasion, identifying both prognostic and predictive biomarkers. We further highlight how spatially resolved analyses have uncovered previously unrecognized axes of communication between cancer cells and key stromal and parenchymal populations that were not apparent from bulk or single-cell molecular analyses alone. These spatially mapped ligand-receptor networks reveal coordinated signaling circuits that define targetable mechanisms of resistance and progression within the microenvironment. Finally, we explore the translational implications where spatial signatures may guide risk stratification, treatment selection, and clinical trial design. By integrating spatial architecture with molecular function, spatial omics is transforming our understanding of liver cancer biology and paving the way for spatially informed precision oncology.
Markaide E, Izquierdo-Sanchez L, Olaizola I
… +15 more, Urretabizkaia-Garmendia J, Ruiz de Gauna M, Buqué X, Lasa-Elosegi I, Lapitz A, Bais T, Val B, Rodrigues PM, Castro RE, Perugorria MJ, Bujanda L, Gansevoort R, Drenth JPH, Aspichueta P, Banales JM
BACKGROUND AND AIM: Polycystic liver disease (PLD) is a hereditary disorder characterized by the progressive development and enlargement of intrahepatic biliary cysts, which can lead to significant morbidity. Liver trans...BACKGROUND AND AIM: Polycystic liver disease (PLD) is a hereditary disorder characterized by the progressive development and enlargement of intrahepatic biliary cysts, which can lead to significant morbidity. Liver transplantation remains the only curative treatment. Mutations in endoplasmic reticulum (ER)-related genes contribute to ER stress in cystic cholangiocytes, promoting disease progression. Given the functional interplay between ER and mitochondria, we investigated mitochondrial dynamics and metabolism in cystic cholangiocytes from both PLD patients and Pkhd1mut rats to identify novel therapeutic targets. APPROACH AND RESULTS: Cystic cholangiocytes exhibited increased mitochondrial mass, membrane potential, and superoxide levels, along with enhanced bioenergetic capacity and ATP production compared with normal cholangiocytes. These alterations were linked to the upregulation of electron transport chain protein complexes. Metabolic reprogramming involved enhanced oxidation of glucose, glutamine, and/or fatty acids, as well as increased denovo cholesterol synthesis and accumulation. Chronic treatment with pravastatin, a cholesterol synthesis inhibitor, significantly reduced hepatomegaly, cyst volume, and fibrosis in Pkhd1mut rats. It also normalized mitochondrial hyperactivity and reduced the proliferation of cystic cholangiocytes in culture, effects that were similarly observed with other statins such as atorvastatin and simvastatin. Importantly, a case-control study in PLD patients showed that statin use was associated with reduced liver growth, further supporting its potential therapeutic role. CONCLUSIONS: Mitochondrial and metabolic dysregulation are central to the pathogenesis of PLD. Targeting cholesterol metabolism with statins represents a promising therapeutic strategy to slow disease progression and reduce cyst burden.
BACKGROUND AND AIMS: Hepatitis delta virus (HDV) is a satellite virus of the hepatitis B virus (HBV), with a single-stranded and rod-like circular RNA encoding only one protein, the hepatitis delta antigen (HDAg). Lackin...BACKGROUND AND AIMS: Hepatitis delta virus (HDV) is a satellite virus of the hepatitis B virus (HBV), with a single-stranded and rod-like circular RNA encoding only one protein, the hepatitis delta antigen (HDAg). Lacking its own replicase, the highly self-complementary HDV RNA hijacks host RNA polymerase II (Pol-II), eliciting a unique and incompletely understood RNA-templated transcriptional activity. Because transcription by Pol-II is regulated at multiple steps by various cyclin-dependent kinases (CDKs), we investigated whether CDKs contribute to HDV replication. APPROACH AND RESULTS: Using selective compounds targeting transcriptional cyclin-dependent kinases (CDKs), we identified the Mediator kinase CDK8 and its paralog CDK19 as key cofactors for HDV replication. Loss of CDK8/19 activity by small-molecule inhibitor MSC2530818 or genetic knockouts completely prevents the establishment of HDV replication in multiple cell culture models and partially suppresses HDV RNA synthesis during the steady-state replication phase. Ectopic expression of the small HDAg, but not its methylation site R13 mutant, restored HDV replication in CDK8/19-deficient cells. Inactivation of CDK8/19 did not alter phosphorylation of small HDAg at Ser177, but was associated with reduced phosphorylation of the C-terminal domain of Pol-II, consistent with impaired transcriptional activity. CONCLUSIONS: Our findings reveal the essential role of CDK8/19 in mediating the transcriptional activity of Pol-II during HDV replication, which is partially counteracted by HDV-encoded small HDAg.
BACKGROUND AND AIMS: Primary sclerosing cholangitis (PSC) is a pre-malignant condition with elevated risk of hepatopancreatobiliary cancers (HPBCa) and colorectal cancer (CRC) when inflammatory bowel disease (IBD) is pre...BACKGROUND AND AIMS: Primary sclerosing cholangitis (PSC) is a pre-malignant condition with elevated risk of hepatopancreatobiliary cancers (HPBCa) and colorectal cancer (CRC) when inflammatory bowel disease (IBD) is present. We described cancer burden in a contemporary PSC-IBD cohort, and assessed rates of subsequent cancers, liver transplant, and death post-colectomy or post-cholecystectomy status. METHODS: Using linked health administrative databases, we calculated cause-specific cumulative incidences of HPBCa-related outcomes (diagnoses/deaths) and non-HPBCa-related transplant/death among individuals with PSC-IBD in Ontario, Canada (2002-2018), followed to 2021. Transition probabilities and transition intensity ratios (TIR) were evaluated using a multistate Markov model. RESULTS: Among 476 individuals with incident PSC-IBD, there was a 54% probability of remaining event-free at 10 years. Approximately 1 in 20 experienced an HPBCa-related outcome, and up to 1 in 4 had a non-HPBCa-related transplant/death. During follow-up, 13% experienced multiple events. Age was associated with HPBCa (HR 1.02, 95% CI 1.01-1.04), but not sex. Mortality occurred more frequently post-colectomy (TIR 3.08, 95% CI 1.7-5.59) and post-cholecystectomy (TIR 3.85, 95% CI 2.21-6.64) relative to event-free PSC-IBD, but there were no differences in post-surgery incidence of cancer or transplant. CONCLUSIONS: While some individuals with PSC-IBD experienced an extended event-free disease course, a large proportion experienced disease-related cancer, colectomy, cholecystectomy, and transplant events. Higher mortality rates observed after surgery were likely related to underlying disease processes that motivated surgical intervention (eg, dysplasia/malignancy, refractory IBD), rather than the surgery itself. Understanding how PSC-IBD disease trajectories vary can inform individual management and patient counselling.
BACKGROUND AND AIMS: Metabolic dysfunction-associated steatotic liver disease (MASLD), including its more severe form, metabolic dysfunction-associated steatohepatitis (MASH), is increasingly recognized as a critical glo...BACKGROUND AND AIMS: Metabolic dysfunction-associated steatotic liver disease (MASLD), including its more severe form, metabolic dysfunction-associated steatohepatitis (MASH), is increasingly recognized as a critical global health challenge. This study investigates the role of hepatic GPR75 in MASH progression. APPROACH AND RESULTS: Although GPR75 is not abundantly expressed in the liver in healthy individuals, its protein levels significantly increase during MASH. Depletion of Gpr75 in either the whole liver or specifically in hepatocytes protected mice from diet-induced hepatic steatosis, while hepatocyte-specific overexpression of Gpr75 exacerbated diet-induced MASH and liver fibrosis. The deficiency of hepatic Gpr75 activated the GNAI2-cAMP-PKA signaling pathway in the livers of MASLD mice, reducing SREBP-1c maturation and de novo lipogenesis. Mechanistically, VPS35 stabilized GPR75 by recycling it to the hepatocyte membrane, thereby decreasing its degradation during MASH progression. CONCLUSIONS: This study demonstrates that GPR75 serves as a novel regulator of MASLD/MASH by modulating hepatic fatty acid metabolism. These findings suggest that GPR75 suppression may represent a potential therapeutic strategy for MASLD/MASH treatment.
BACKGROUND AND AIMS: Liver fibrosis poses a major health threat globally, with an acute shortage of effective treatments to stop or reverse its progression. Hepatocyte injury serves as the primary cause of liver fibrosis...BACKGROUND AND AIMS: Liver fibrosis poses a major health threat globally, with an acute shortage of effective treatments to stop or reverse its progression. Hepatocyte injury serves as the primary cause of liver fibrosis, and restoring the expression of damaged proteins via mRNA delivery represents a promising therapeutic strategy. Clinical studies and animal models have demonstrated that mitochondrial ornithine transcarbamylase ( OTC ) deficiency correlates with increased incidence of liver fibrosis. In this study, we examined the therapeutic potential of OTC mRNA delivery for liver fibrosis. APPROACH AND RESULTS: Analysis of OTC expression and distribution in liver fibrosis revealed significant downregulation in fibrotic tissues. Restoring OTC expression in hepatocytes through AAV8-TBG-OTC significantly inhibits the progression of liver fibrosis. OTC mRNA sequences were developed using a codon-optimization artificial intelligence (AI) tool and synthesized with N1-methylpseudouridine modification, followed by encapsulation into lipid nanoparticle (LNP). Repeated in vivo delivery of OTC mRNA-LNP induced a robust inhibition of fibrogenesis in multiple mouse liver fibrosis models. Mechanistically, restoring OTC mRNA expression significantly recovers impaired mitochondrial and hepatocyte functions, while concurrently inhibiting paracrine ammonia-induced activation of HSCs and macrophages. OTC mRNA significantly suppresses collagen deposition, while the THR- β agonist Resmetirom potently inhibits lipid accumulation; the combination exhibits potent complementary efficacy against metabolic dysfunction-associated steatohepatitis (MASH) liver fibrosis. CONCLUSIONS: Collectively, our study provides the first direct preclinical evidence supporting OTC mRNA therapeutics for treating liver fibrosis and establishes proof-of-concept for mRNA therapy as a novel strategy to reverse liver fibrosis.
BACKGROUND AND AIMS: Cholangiocarcinoma (CCA) is a rare but aggressive malignancy with poorly understood genetic susceptibility. To date, genome-wide association studies (GWAS) investigating germline variants associated...BACKGROUND AND AIMS: Cholangiocarcinoma (CCA) is a rare but aggressive malignancy with poorly understood genetic susceptibility. To date, genome-wide association studies (GWAS) investigating germline variants associated with CCA risk remain limited. We aimed to identify genetic risk loci for CCA and its clinical subtypes through comprehensive GWAS and post-GWAS analyses. APPROACH AND RESULTS: We conducted a GWAS of 2366 CCA cases and 11,750 controls of European ancestry. Genome-wide significant loci ( p <5×10 -8 ) were identified and further examined through fine-mapping, functional annotation, and HLA imputation. Subgroup analyses were conducted by CCA subtypes and primary sclerosing cholangitis (PSC) status. Cross-trait linkage disequilibrium score regression and Mendelian randomization were employed to investigate the shared genetic architecture and potential causal relationships with a diverse range of traits. We identified 1 new genome-wide significant variant, rs535777 (OR=1.44), near HLA-DRB1/DQA1 associated with CCA, and 2 variants associated with extrahepatic CCA: rs116224263 (OR=0.17) in LINC02506 at 4p15.1 and rs6914950 (OR=1.63) near HLA-DRB1/DQB1 . Stratified analyses revealed rs2395184 (OR=3.51) near HLA-DRA/DRB5 associated with PSC-related CCA, and rs142674434 (OR=2.98) in THSD7A at 7p21.3 associated with non-PSC-related CCA. HLA imputation uncovered new amino acid residues associated with disease risk. Cross-trait analyses identified shared genetic signals between CCA and anthropometric, lipidemic, lifestyle, and medical traits. Mendelian randomization supported putative causal associations for 12 traits with CCA or its subtypes. CONCLUSIONS: Our large-scale GWAS highlights new genetic variants and HLA-linked mechanisms underlying CCA susceptibility. Integrating multi-step post-GWAS approaches enhances understanding of CCA pathogenesis and may facilitate the development of risk biomarkers for early detection and precision prevention strategies.
BACKGROUND AND AIMS: Older age increases susceptibility to metabolic dysfunction-associated steatotic liver disease (MASLD), but whether it impacts response to therapies, and how the therapies impact regulators of biolog...BACKGROUND AND AIMS: Older age increases susceptibility to metabolic dysfunction-associated steatotic liver disease (MASLD), but whether it impacts response to therapies, and how the therapies impact regulators of biological aging, are poorly understood. Statins inhibit the mevalonate pathway to block cholesterol biosynthesis and are widely used in MASLD patients to reduce cardiovascular disease. Whether statins prevent progression to cirrhosis is under investigation. However, the molecular effects of statins in human liver, particularly in the context of aging, remain poorly defined. APPROACH AND RESULTS: We analyzed liver transcriptomes and matched clinical data from 368 adults enrolled in the Duke MASLD Biorepository with a focus on age-dependent responses and the interplay between senescence and ferroptosis, a regulated death process that is constrained by the mevalonate pathway. Serum ALT, AST, and LDL cholesterol levels were lower in statin users of both sexes, particularly among older individuals. Transcriptome analyses revealed that statin use is strongly associated with suppression of senescence-related pathways. Statin use is also associated with increased activation of pathways linked to ferroptosis. Both responses persisted after propensity score matching to control for clinical confounders and were validated in an independent obese cohort. CONCLUSIONS: Age-dependent transcriptional remodeling in the liver differs in statin users and non-users. Pathways involved in senescence are suppressed while those that promote ferroptosis are induced in statin users. These results suggest that statins may suppress biological aging in MASLD by acting as senolytics and highlight the complex, context-specific roles of senescence in liver adaptation and remodeling.
BACKGROUND AND AIMS: Statins have been investigated for their potential to reduce liver-related complications in chronic liver diseases, but evidence in primary biliary cholangitis (PBC) remains limited. This study aimed...BACKGROUND AND AIMS: Statins have been investigated for their potential to reduce liver-related complications in chronic liver diseases, but evidence in primary biliary cholangitis (PBC) remains limited. This study aimed to assess the association between statin use and the risk of hepatic decompensation using a target trial emulation (TTE) design. APPROACH AND RESULTS: We performed a sequential TTE using 2 electronic health record databases: Mass General Brigham (MGB, Boston, USA) and Asan Medical Center (AMC, Seoul, Korea). Adults diagnosed with PBC between 2001 and 2024 were eligible. Statin use was defined as a cumulative duration of ≥90 days. In each monthly trial, statin initiators were matched 1:2 to non-users using propensity score matching. The primary outcome was hepatic decompensation; the secondary outcome was a composite major adverse liver outcome (MALO), including decompensation, hepatocellular carcinoma, and liver transplantation. Among 2889 eligible patients, 443 statin users were matched to 886 non-users. Over a median follow-up of 3.8 years, hepatic decompensation occurred in 24 statin users (5.4%) and in 67 non-users (7.6%) [hazard ratio (HR), 0.61; 95% confidence interval (CI): 0.38-0.97]. Statin use was also associated with a reduced risk of MALO (HR, 0.58; 95% CI: 0.38-0.89). Sensitivity analyses stratified by data source (MGB, HR 0.65; AMC, HR 0.60) and cirrhosis status (HR 0.70 for cirrhosis; HR 0.57 for without) showed similar directional trends. CONCLUSIONS: Statin use was consistently associated with a lower risk of hepatic decompensation and major liver events in patients with PBC, supporting a potential protective effect.
Primary liver cancer, mainly HCC and intrahepatic cholangiocarcinoma, remains a leading cause of cancer mortality worldwide. Early detection is crucial for curative treatment, yet current surveillance and diagnostic stra...Primary liver cancer, mainly HCC and intrahepatic cholangiocarcinoma, remains a leading cause of cancer mortality worldwide. Early detection is crucial for curative treatment, yet current surveillance and diagnostic strategies, primarily ultrasound surveillance and contrast-enhanced CT or MRI, suffer from operator dependence, limited sensitivity, and interpretive variability. Artificial intelligence (AI) offers transformative potential across the liver cancer continuum, from surveillance to diagnosis and pathology. Deep learning-based models have improved ultrasound detection of small liver tumors, enabling automated triage and reducing workload. On CT and MRI, AI systems achieve expert-level performance for lesion detection, segmentation, and characterization, supporting standardized interpretation through frameworks such as liver imaging reporting and data system. In digital pathology, AI algorithms can distinguish between HCC and cholangiocarcinoma, classify dysplastic nodules, and even predict future cancer development from biopsy slides. Recent advances in foundation models and multimodal AI promise to unify radiology, pathology, and molecular data, paving the way for comprehensive, patient-specific disease modeling. However, widespread clinical integration faces major challenges, including data privacy, regulatory approval, cost sustainability, and algorithmic bias. Large, prospective multicenter validation studies are essential to confirm clinical benefit and safety. Ultimately, the careful implementation of trustworthy and explainable AI tools could enable earlier detection, greater diagnostic precision, and more equitable liver cancer care.
BACKGROUND AND AIMS: Type 2 diabetes mellitus (T2DM) is a recognized modifiable risk factor for HCC and liver-related mortality. The effects of newer antidiabetic agents-including glucagon-like peptide-1 receptor agonist...BACKGROUND AND AIMS: Type 2 diabetes mellitus (T2DM) is a recognized modifiable risk factor for HCC and liver-related mortality. The effects of newer antidiabetic agents-including glucagon-like peptide-1 receptor agonists (GLP-1 RAs), sodium-glucose cotransporter-2 (SGLT-2) inhibitors, and dipeptidyl peptidase-4 (DPP-4) inhibitors-on hepatic outcomes remain uncertain. We aimed to evaluate whether these therapies reduce the risk of HCC and non-HCC liver-related events (LREs) in patients with T2DM. APPROACH AND RESULTS: A systematic literature search was performed to identify studies reporting hepatic complications among patients with T2DM prescribed GLP-1 RAs, SGLT-2 inhibitors, or DPP-4 inhibitors. Comparisons were made against patients receiving various glucose-lowering therapies other than the drug of interest. Subgroup analyses were conducted in patients with chronic liver disease. Random-effects meta-analyses were used to estimate pooled hazard ratios (HRs). Of 2,228 records screened, 36 cohort studies comprising 5,363,858 patients were included. Compared with other glucose-lowering therapies, GLP-1 RAs were associated with significantly reduced risks of HCC (pooled HR 0.77, 95% CI [0.66-0.90]) and LREs (0.79 [0.65-0.95]). SGLT-2 inhibitors similarly conferred protection against HCC (0.76 [0.67-0.86]) and LREs (0.82 [0.73-0.92]). By contrast, DPP-4 inhibitors were not associated with hepatoprotection, showing neutral effects on HCC (1.12 [0.91-1.39]) and increased risk of LREs (1.24 [1.15-1.34]). In patients with chronic liver disease, GLP-1 RAs were uniquely associated with reduced hepatic decompensation (0.79 [0.71-0.88]). CONCLUSIONS: GLP-1 RAs and SGLT-2 inhibitors were associated with hepatoprotective effects compared with other glucose-lowering therapies in patients with T2DM, with GLP-1 RAs showing additional benefits in chronic liver disease. These findings provide evidence on the relationships between antidiabetic drug classes and liver-related outcomes in patients with T2DM and may inform clinical decision-making.