Clinical and Molecular Hepatology最新文献

Background and aims: Wilson's disease (WD) is an autosomal recessive disorder of copper metabolism caused by ATP7B mutations. In this study, a novel gene therapy for WD was developed, and its efficacy and safety were evaluated in relevant animal models.

Methods: Codon-optimized full-length or truncated ATP7B (tATP7B) genes were assembled with liver-specific mini promoters to construct an adeno-associated virus serotype 8 vector (AAV8). The expression and activity of these vectors were evaluated in HepG2 cells. AAV8-tATP7B viral particles were produced using a triple-plasmid cotransfection system under good manufacturing practice conditions. Long-term efficacy was evaluated in Atp7b-/- mice at three doses (5×10¹¹, 5×10¹², and 1×10¹³ vg/kg). Single-dose toxicity was assessed over 13 weeks in Sprague-Dawley rats and cynomolgus macaques.

Results: In HepG2 cells, the ability of the truncated ATP7B to export copper was comparable to that of the full-length protein, but the expression efficiency was greater. Alkaline gel electrophoresis confirmed its better compatibility with AAV packaging limits while maintaining genomic integrity, supporting its selection for AAV8-tATP7B production. In a 24-week study in Atp7b-/- mice, AAV8-tATP7B restored copper homeostasis and liver function in a dose-dependent manner and significantly reversed existing liver injury. Toxicity studies in Sprague‒Dawley rats and cynomolgus monkeys revealed no systemic toxicity, whereas reversible liver changes were observed in cynomolgus monkeys at high doses; thus, 6×10¹³ vg/kg was established as the maximum tolerated dose.

Conclusions: These results establish the efficacy and safety profile of AAV8-tATP7B and provide the rationale for its clinical translation in patients with WD.

Hepatocellular carcinoma (HCC) is the most common primary liver cancer and a leading cause of cancer mortality worldwide. Its pathogenesis reflects a combination of tumour-intrinsic heterogeneity and a profoundly immunosuppressive tumour microenvironment (TME). Growing evidence shows that tumours recapitulate developmental programs to establish an oncofetal ecosystem, characterised by the re-expression of foetal antigens and foetal-like stromal and immune subsets. These features drive immune evasion and shape therapeutic response, contributing to immunotherapy outcomes in clinic. This review outlines mechanistic insights into oncofetal reprogramming across tumour, stromal, and immune compartments and evaluates therapeutic strategies that target these dependencies. We highlight emerging vaccine platforms, cellular therapies, and biologics targeting oncofetal antigens, with particular emphasis on mRNA-lipid nanoparticle (LNP) vaccines and their potential to induce robust, durable antitumour immunity. We further discuss rational combinatorial strategies that integrate vaccines with immune checkpoint inhibitors. Finally, we discuss how overcoming liver tolerance and antigenic heterogeneity will be essential for effective oncofetal-directed therapies. Collectively, targeting the oncofetal ecosystem through coordinated vaccine, cellular, and immunotherapeutic strategies offers a path toward more durable responses and broader immunotherapy benefits in HCC.

Background/aims: The GALAD (Gender, Age, Lens culinaris agglutinin-reactive alpha-fetoprotein [AFP-L3], alpha-fetoprotein [AFP], and des-γ-carboxy prothrombin [DCP]) score, widely used for hepatocellular carcinoma (HCC) detection, was primarily derived from cohorts with advanced-stage tumors and elevated biomarker levels, potentially overestimating accuracy in early-stage disease. Furthemore, the lectin-based AFP-L3 assay has poor sensitivity at low AFP concentrations, limiting detection of small or AFP-negative tumors.

Methods: We developed GAFAD, a multivariable model replacing AFP-L3 with fucosylated AFP percentage (AFP-Fuc%), quantified by a validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay. The model was trained and tested using a hepatitis B virus (HBV)-related cohort (HCC n=235; non-HCC n=290), a diagnostically challenging set with substantial overlap in biomarker levels between HCC and non-HCC. Moreover, a final model (GAFAD) was validated in two independent cohorts (HCC n=210, non-HCC n=245), comprising HBV-, HCV-related and non-viral etiologies.

Results: In the development cohort, GAFAD showed superior diagnostic performance to GALAD for distinguishing HCC from non-HCC, with a higher area under the receiver operating characteristic curve (AUC, 0.938 vs. 0.887; p<0.0001) and greater sensitivity (82% vs. 66%) and accuracy (86% vs. 79%) at 90% specificity. In the external validation cohort, GAFAD similarly outperformed GALAD, achieving a higher AUC (0.874 vs. 0.841, p<0.05), greater sensitivity (72% vs. 57%), and improved accuracy (82% vs. 75%) at 90% specificity. This superiority extended to early-stage, very-early-stage, and AFP-negative HCC.

Conclusions: GAFAD provides a reliable and generalizable tool for early HCC detection across diverse etiologies, supporting its clinical applicability in surveillance and diagnosis.

Background: Biliary tract cancer (BTC) frequently harbors KRAS mutations, which are associated with resistance to traditional treatment and a poor prognosis. Synthetic lethality (SL) strategy may provide other targets of KRAS. Therefore, we aim to identify and validate potential therapeutic target of KRAS for the treatment of BTC via SL.

Methods: The dependency (DepMap) projects were used to predict the synthetic lethal gene of KRAS. FDA-approved anticancer drug library was applied to screen potential drugs effective against KRAS-mutant BTC. Furthermore, the synthetic lethal effects or corresponding mechanisms of potential genes and drugs on BTC was investigated using KRAS-mutant and KRAS-wild type BTC cell lines, patient-derived xenografts (PDX), KRAS oncogene-driven tumor models, as well as other KRAS-mutant cancer cell lines.

Results: Initially, we discovered that the loss of GATA2 reduced the viability of KRAS-mutant but not KRAS-wild-type BTC. Subsequently, the drug library screened out disulfiram, which primarily exerts a synthetic lethal effect by inhibiting IL-1β in KRAS-mutant BTC. Mechanistically, GATA2 specifically enhanced the transcription of IL-1β to promote NF-κB signaling in KRAS-mutant BTC. IL-1β inhibition phenocopied GATA2 deficiency, leading to reduced KRAS-mutant BTC viability. These synthetically lethal effects were confirmed using PDX, a KRAS oncogene-driven tumor model, as well as in other KRAS-mutant cancer cell lines.

Conclusions: In summary, these results indicate that inhibiting GATA2/IL1β could be a therapeutic strategy in KRAS-mutant BTC and potentially other cancers.

Severe alcohol-associated hepatitis (SAH) is the most aggressive form of alcohol-associated liver disease and is associated with very high short-term mortality. It is characterized by the acute onset of jaundice in the context of ongoing alcohol use, most commonly defined by a Maddrey Discriminant Function ≥32 or a Model for End-Stage Liver Disease score ≥ 20. Despite its increasing global burden and substantial healthcare costs, therapeutic options remain limited, and outcomes are poor. The severity of liver failure, systemic inflammation, infectious complications, and extrahepatic organ dysfunction determines the prognosis in SAH. The pathophysiology of SAH is multifactorial, involving direct hepatotoxicity from alcohol metabolites, oxidative stress, dysregulated immune activation, gut dysbiosis with increased intestinal permeability, impaired hepatic regeneration, and genetic susceptibility. These interrelated mechanisms culminate in an exaggerated inflammatory response driven by macrophage activation and cytokine release, resulting in hepatocellular injury and multi-organ failure. Glucocorticoids remain the guideline-recommended standard of care for selected patients; however, their benefit is limited to modest short-term survival gains, with high rates of non-response and infection. Numerous investigational therapies targeting inflammation, oxidative stress, liver regeneration, bile acid signaling, epigenetic regulation, and the gut-liver axis have been evaluated, with largely disappointing results. Emerging approaches, including interleukin-22 agonists and epigenetic modulators such as larsucosterol, show promise but require validation in well-designed trials. This review synthesizes current evidence on the definition, prognostic assessment, and pathophysiology of SAH, critically appraises existing and emerging therapies, and highlights the need for combination strategies, improved patient stratification, and personalized treatment approaches.