BACKGROUND AND AIMS:Pradefovir mesylate is a novel liver-targeting prodrug of adefovir developed using HepDirect technology. We report that the phase 3 trial is to compare the efficacy and safety of pradefovir versus tenofovir disoproxil fumarate (TDF) in patients with chronic hepatitis B (CHB). APPROACH AND RESULTS:This ongoing randomized, double-blind, non-inferiority, phase 3 study was conducted at 58 sites in China. Patients with CHB were randomly assigned (2:1) to receive either 45 mg daily pradefovir or 300 mg daily TDF with a matching placebo. The primary efficacy endpoint was defined as the proportion of patients whose HBV DNA level was <29 IU/mL at week 48. All participants who received at least 1 dose of the study drug were included in efficacy and safety analyses with pre-specified renal and bone endpoints at week 96. A total of 1170 patients were screened during the study, and 908 patients received the study drug. At week 48, viral suppression (HBV DNA <29 IU/mL) rates of pradefovir were non-inferior to TDF in HBeAg-positive [-1.8% (95% CI: -9.7 to 6.1)] and HBeAg-negative [2.6% (95% CI: -5.1 to 10.3)] patients, using a non-inferiority margin of -12%. By week 96, HBeAg-positive patients showed significantly greater HBsAg decline (≥1 log 10  IU/mL) with pradefovir (39.3% vs. 29.9%). Pradefovir demonstrated a more favorable safety profile at week 96, including significantly fewer drug-related adverse events (58.6% vs. 71.9%), improved bone/renal safety, and a lower incidence of elevated creatine phosphokinase-MB. CONCLUSIONS:Pradefovir should be a highly recommended treatment option for adult patients with CHB.

BACKGROUND:Chronic HBV and HDV coinfection represents the most severe form of viral hepatitis. This study aimed to assess the safety and efficacy of libevitug (HH-003), a novel human monoclonal antibody targeting the PreS1 domain of the HBV large envelope protein, in patients with chronic HBV and HDV coinfection. METHODS:This single-center, open-label, proof-of-concept phase IIa clinical trial enrolled 9 participants who received i.v. infusion of 20 mg/kg libevitug every 2 weeks for 24 weeks, followed by a 24-week follow-up period. RESULTS:At week 24, 8 participants (88.9%) exhibited a reduction in HDV RNA ≥1 log10 from baseline, with 4 (44.4%) achieving undetectable levels. 7 participants (77.8%) achieved virological response (undetectable HDV RNA or ≥2 log10 decline from baseline). Among the 5 participants with elevated baseline ALT, 3 (60.0%) achieved a combined virological and biochemical response (ALT normalization). At week 48, 6 participants (66.7%) maintained the virological response achieved at the end of treatment (week 24), with 3 (33.3%) maintaining undetectable HDV RNA levels. Among the 7 participants with quantifiable HBV DNA at baseline, 2 (28.6%) achieved undetectable HBV DNA or ≥1 log10 reduction at week 24. Serum HBsAg levels remained unchanged throughout the study period. Treatment-emergent adverse events were reported in 8 participants (88.9%), with only 1 participant (11.1%) experiencing treatment-related adverse events. No grade 3 or serious adverse events were reported, and no discontinuations due to adverse events occurred. CONCLUSIONS:Libevitug demonstrated promising antiviral activity, achieving substantial HDV RNA reduction, a high rate of ALT normalization, and favorable safety and tolerability in participants with chronic HBV and HDV coinfection.

BACKGROUND:Rifaquizinone (RQZ, TNP-2092) is a novel drug for treating hepatic encephalopathy (HE). This study assessed the safety, pharmacokinetics (PK), and efficacy of RQZ capsules in healthy participants and cirrhosis patients with hyperammonemia. RESEARCH DESIGN AND METHODS:We conducted three double-blind, randomized, placebo-controlled trials. Trials 1 and 2 enrolled healthy participants, while Trial 3 involved 36 cirrhosis patients with hyperammonemia. RESULTS:In healthy participants and liver cirrhosis patients with hyperammonemia, RQZ was safe and well tolerated. Adverse events were generally mild or moderate. Both food and liver cirrhosis disease status increased the exposure of RQZ, which was around 1% of that of the corresponding doses after intravenous administration. The effect in lowering blood ammonia was dose-dependent. The changes from baseline in blood ammonia levels at day 15 were -14.1 and 0.5 μmol/L in 600 mg and placebo cohort. The percentage of achieving normal blood ammonia during treatment were 31.9% and 7.4% in 600 mg and placebo cohort. CONCLUSIONS:Although this study enrolled a relatively small cohort of cirrhotic patients with hyperammonemia, RQZ demonstrated an excellent safety profile as an intestinally restricted agent and showed promising potential for lowering blood ammonia and treating hepatic encephalopathy.

BACKGROUND:ALG-000184 (pevifoscorvir sodium) is a prodrug of the class-E (empty) capsid assembly modulator (CAM), ALG-001075. CAM-Es prevent hepatitis B virus (HBV) replication by inhibiting capsid assembly and pregenomic RNA encapsidation and preventing covalently closed circular DNA (cccDNA) establishment. This part of the study aimed to assess the safety, tolerability, pharmacokinetics and short-term antiviral activity of ALG-000184 in participants with HBeAg-negative and HBeAg-positive chronic HBV infection to determine the subsequent development pathway for this novel therapeutic. METHODS:ALG-000184-201 was a phase 1, multicentre, randomised, dose escalation trial designed to evaluate the safety and pharmacokinetics of ALG-000184 in healthy participants (parts 1-2, reported separately) and antiviral activity in treatment-naive or currently not treated participants with chronic HBV infection (part 3, presented here). Eligible participants were 18-65 years old with BMI 18·0-35·0 kg/m2, and HBeAg-negative or HBeAg-positive chronic HBV infection with serum HBV DNA concentration more than 2000 IU/mL and serum ALT concentrations less than 5 times the upper limit of normal. Eligible patients were randomly assigned (4:1) to receive 28 days of oral ALG-000184 (10 mg, 50 mg, 100 mg, or 300 mg) or matching placebo once per day. Randomisation was managed via an interactive web response system and other than knowing the dose being evaluated in each cohort, all investigational site personnel and study participants were unaware of treatment assignments. The primary objective of this study was to evaluate the safety and tolerability of ALG-000184 in all randomly assigned participants (intention to treat population) using safety data including all treatment emergence adverse events, physical exam and vital signs findings, 12-lead electrocardiogram, and clinical laboratory findings. This study was registered at ClinicalTrials.gov, NCT04536337, and is complete. FINDINGS:Between March 16, 2021, and Oct 13, 2022, 135 participants were screened for inclusion from seven clinical investigational sites in Hong Kong, China, Moldova, New Zealand, and the UK. 76 participants did not meet one or more of the inclusion criteria or met one or more exclusion criteria and 59 participants were enrolled and randomly assigned to receive ALG-000184 10 mg (n=14), 50 mg (n=8), 100 mg (n=16), or 300 mg (n=10) or placebo (n=11). Participants had a median age of 37·0 years (IQR 31·0-45·5). 30 (51%) of 59 patients were male and 29 (49%) were female. 40 (68%) of patients were Asian. 28 (58%) of 48 participants assigned to receive ALG-000184 and seven (64%) of 11 in the placebo group experienced at least one treatment-emergent adverse event (TEAE). The most common TEAEs were increased alanine aminotransferase (ALT; 15 [31%] of 48 participants assigned to ALG-000184 and two [18%] of 11 participants assigned to placebo), increased aspartate aminotransferase (AST) concentrations (12 participants [25%] vs one [9%]), hyperuricemia (five [10%] vs two [18%]), headache (four [8%] vs one [9%]), elevated amylase (three [6%] vs none) and upper respiratory tract infection (three [6%] vs none). There were two serious adverse events unrelated to ALG-000184 (mild spinal pain and spontaneous pneumothorax). There were no deaths in the study. Doses of 10 mg to 300 mg ALG-000184 for 28 days resulted in multi-log reduction of HBV DNA (about 3-4 log10 IU/mL) and HBV RNA (about 0·5-3·0 log10 copies per mL). 19 (83%) of 23 HBeAg-negative participants achieved HBV DNA levels less than the lower limit of quantification (10 IU/mL) and 23 (100%) achieved HBV RNA levels less than the lower limit of quantification (10 copies per mL). Due to higher baseline levels, no HBeAg-positive participant achieved HBV DNA or HBV RNA less than the lower limit of quantification by 28 days. A decrease in HBsAg of 0·5 log10 IU/mL was observed in one HBeAg-positive participant who received 100 mg ALG-000184 and four of eight HBeAg-positive participants who received 300 mg ALG-000184 for 28 days had decreases of 0·2-0·8 log10 IU/mL. ALG-000184 had a dose-dependent linear pharmacokinetic profile. INTERPRETATION:ALG-000184 was safe and well tolerated, demonstrated predictable pharmacokinetic properties, and reduced HBV DNA and HBV RNA at all doses regardless of HBeAg status. These results support the evaluation of 300 mg ALG-000184 over longer periods in larger studies to determine its potential role in chronic suppressive therapy, with or without a nucleos(t)ide analogue, or as part of a finite curative regimen. FUNDING:Aligos Therapeutics.

Laser desorption/ionization mass spectrometry (LDI-MS) is attractive for serum and spatial metabolomics, but metabolite analysis is limited by inefficient ionization and severe low-mass interference from conventional matrices. Here we report a vacancy-clustered molybdenum oxide (MoO x ) matrix for LDI-MS analysis of serum profiling and spatial tissue imaging. Vacancy-clustered defects, rather than vacancy-lean, point-defect-dominated, or over-reduced states, markedly enhance photogenerated charge separation and interfacial charge transport, enabling efficient metabolite ionization. This highlights vacancy organization as a key design fundamental for inorganic LDI matrices. Compared with conventional organic matrices, vacancy-clustered MoO x increases desorption and ionization efficiency by 3- to 27-fold and expands metabolite coverage by at least twofold. In-situ characterizations and calculations reveal that vacancy-clustered defect organization redirects post-excitation surface charge migration, promotes proton-coupled interfacial activation and lowers the energetic barrier for metabolite-ion release, establishing a charge-driven desorption and ionization basis for inorganic LDI matrices. Leveraging these advantages, this unified LDI-MS platform enables high-sensitivity cohort-scale serum metabolomics and high-resolution spatial tissue metabolomics in hepatocellular carcinoma. It reveals circulatory metabolic characteristics spatially associated with tumor regions, providing direct evidence of metabolic associations between blood and tumor tissue. These findings show vacancy-clustered engineering as a practical strategy for inorganic LDI matrices and provide an integrated platform for multidimensional disease metabolomics.

BACKGROUND & AIMS:Spontaneous hepatitis B surface antigen (HBsAg) clearance is rare in chronic hepatitis B (CHB). Identifying predictors of antiviral responses is needed. METHODS:We retrospectively identified 1583 treatment-naive children with CHB confirmed by liver biopsy at the Fifth Medical Center of PLA General Hospital from January 2007 to April 2024. Subsequently, 852 patients received antiviral therapy and were followed up every 3 to 6 months. The median follow-up time was 3.5 years. Virological, pathological, and noninvasive tests were performed to explore predictors of antiviral response. RESULTS:Patients in the HBsAg loss group were younger than those in the nonloss group, with a median age of 3 vs 7 years (P < .001). There were 118 (60.8%), 168 (48.3%), 26 (16.7%), and 12 (7.8%) children with CHB who achieved HBsAg clearance among those aged <3, 3 to 6, 7 to 12, and 13 to 18 years (P < .001). The hepatitis B envelope antigen (HBeAg) loss rate in children aged <3 years (83.6%) was higher than that in older children (P < .001). Compared with older children, children younger than 3 years needed a shorter median time to achieve HBsAg and HBeAg loss (41.4 and 47.4 weeks, respectively) (all P < .001). Notably, children younger than 3 years had a lower median liver stiffness measurement value (5.0 kPa) during the follow-up period, with a shorter median time (75.9 weeks) to achieve liver stiffness measurement regression (all P < .001). CONCLUSIONS:Initiating treatment at a younger age predicts more positive and rapid hepatitis B virus clearance in children with CHB. Early intervention also improves hepatic elastography parameters in children with CHB.

The capacity to sense mechanical stimuli represents one of the most fundamental characteristics of life, enabling organisms to navigate their environment. Here, we identify the mechano-antiviral response system (MARS), a Piezo1-mediated pathway that confers broad-spectrum antiviral immunity distinct from known innate immune systems. Using enterovirus D68 (EV-D68) as a model, we demonstrate that cellular compression or fluid pressure activates Piezo1-dependent antiviral resistance in non-immune cells. Piezo1 functions as a natural antiviral factor, and its pharmacological activation protects against multiple clinical isolates of EV-D68. Mechanistically, the activation of the biomechanical-Piezo1 axis results in a marked reduction in host cell membrane fluidity, a critical determinant for viral entry. Consequently, MARS restricts the replication of diverse viruses, including rhinovirus and influenza. In vivo studies reveal that Piezo1 agonists or mechanical stimuli alleviate EV-D68-induced neurological damage and lethality. Our findings underscore MARS-mediated membrane remodeling as a non-canonical antiviral strategy, expanding the paradigms of immune stimulation.

AHB-137 is a novel antisense oligonucleotide targeting a conserved 3’-terminal region of HBV mRNA. This phase 1a/1b study evaluated the safety, pharmacokinetics (PK), and antiviral activity of AHB-137 in Chinese healthy volunteers (HVs) and chronic hepatitis B (CHB) patients. In Phase 1a, 52 HVs received single ascending doses (75–450 mg) or multiple ascending doses (MAD; 150 or 300 mg) of AHB-137 or placebo. In Phase 1b, 20 HBeAg-negative, nucleos(t)ide analogue (NA)-suppressed CHB patients with baseline HBsAg 100–1000 IU/mL received MAD AHB-137 (150 or 300 mg) or placebo (8:2), and 2 patients with baseline HBsAg 1000–3000 IU/mL received open-label AHB-137 300 mg. MAD comprised 6 subcutaneous doses over 4 weeks including loading doses on Days 4 and 11, with safety, PK and virologic follow-up assessments. No serious adverse events, deaths or discontinuations occurred. Most treatment-related adverse events were Grade 1–2, including injection site reactions, pyrexia, and asymptomatic lab abnormalities. Transaminase elevations were self-limiting; one Grade 3 ALT elevation in the 300 mg CHB cohort coincided with rapid HBsAg loss. PK showed rapid absorption (Tmax 3–5 h), dose-proportionality, and a terminal half-life of 128–195 h without meaningful accumulation. In CHB patients, 4 weeks of AHB-137 induced dose-dependent HBsAg reductions (mean maximum − 1.1 vs. − 2.1 log10 IU/mL at 150 vs. 300 mg), 50.0

BACKGROUND & AIMS:Acetaminophen (APAP) overdose is the leading cause of acute liver failure in Western countries. However, the inflammatory mechanisms underlying APAP-induced liver injury (AILI) remain poorly understood. Interleukin (IL)-19 exerts its biological effects through binding to the IL-20 receptor complex (IL-20R1/IL-20R2). This study aimed to investigate the role of IL-19 in AILI. METHODS:Il19-deficient mice and myeloid cell-specific Il20r2 knockout mice were generated and subjected to AILI. Single-cell RNA sequencing (ScRNA-seq) was performed to analyze hepatic macrophage subsets. RESULTS:Il19-deficient mice exhibited increased susceptibility to AILI, accompanied by a distinct transcriptional profile and a marked increase in proinflammatory macrophage infiltration. Treatment with recombinant IL-19 significantly suppressed proinflammatory macrophage infiltration and attenuated AILI. scRNA-seq analysis revealed that myeloid cell-specific Il20r2 deficiency exacerbated AILI and was associated with increased infiltration of S100A8/A9+ proinflammatory macrophages. Mechanistically, IL-19 downregulated CCAAT/enhancer-binding protein β (C/EBPβ) expression in macrophages by promoting its post-translational SUMOylation. This reduced C/EBPβ-mediated transcription of proinflammatory genes, including those regulating S100A8/A9 expression, thereby ameliorating AILI. CONCLUSIONS:IL-19 plays a key role in limiting proinflammatory macrophage infiltration and thereby ameliorates early-stage AILI. These findings suggest that IL-19 may represent a promising therapeutic target for AILI. IMPACT AND IMPLICATIONS:Although IL-19 signals through the IL-20 receptor complex (IL-20R1/IL-20R2), its role in liver disease remains incompletely understood. Here, we show that IL-19 protects against APAP-induced liver injury by restricting the infiltration of S100A8/A9+ proinflammatory macrophages. Both Il19-deficient mice and myeloid cell-specific Il20r2-deficient mice displayed increased susceptibility to APAP hepatotoxicity and enhanced accumulation of S100A8/A9+ proinflammatory macrophages. Mechanistically, IL-19 promotes C/EBPβ SUMOylation, thereby suppressing proinflammatory gene expression. These findings identify IL-19 as a potential therapeutic target for acute liver injury.

The objective of this study was to investigate the efficacy and safety of oral tolvaptan for hyponatremia in patients with cirrhosis in China. In this post hoc subgroup analysis of a phase 2 clinical trial of oral tolvaptan for hyponatremia due to cirrhosis, heart failure, and syndrome of inappropriate antidiuretic hormone secretion in China, patients with hyponatremia due to cirrhosis received placebo or tolvaptan for 7 days (15 mg titrated to 30 or 60 mg/day). The primary endpoint was the average daily change in serum sodium level from baseline to days 4 and 7. We enrolled 131 patients (90 males, 41 females) with cirrhosis mainly due to CHB (70.2

BackgroundHepatocellular carcinoma (HCC) arises in the liver, an organ with active lipid metabolism, and is accompanied by marked myeloid-cell remodeling. Whether lipid-handling macrophage states are enriched in HCC and how they relate to candidate ligand–receptor communication and tumor microenvironmental remodeling remain unclear.MethodsWe analyzed paired tumor and adjacent liver single-cell transcriptomic data from 10 HCC patients. Tumor-associated lipid-handling candidates were screened using immune-cell composition, metabolic pathway activity, and tumor-upregulated lipid-related gene programs, with APOE prioritized for downstream analysis. Myeloid reclustering, module scoring, and CellChat analysis were used to characterize APOE-positive macrophages and infer APOE-positive macrophage-centered candidate communication. Hepa1–6 subcutaneous tumor models, an Nras-Myc-driven liver tumor model, mouse single-cell transcriptomics, and bone marrow-derived macrophage lipid-loading experiments were used to assess host Apoe deficiency, tumor-cell Apoe perturbation, and macrophage neutral lipid accumulation.ResultsHCC tumors showed altered immune-cell composition and immune-metabolic pathway activity compared with adjacent liver tissues. Among tumor-upregulated lipid-related candidates, APOE was prioritized because of its tumor-associated expression, consistent ranking across lipid-related screening strategies, and functional relevance to lipoprotein and cholesterol handling. APOE-positive macrophages were enriched in tumors and displayed lipid-routing, cholesterol-handling, endolysosomal, and redox-adaptive programs. CellChat analysis identified candidate ligand–receptor interactions involving APOE-positive macrophages, including an APOE–TREM2-related incoming interaction and SPP1–integrin/CD44 outgoing interactions. In the immunocompetent Hepa1–6 model, host Apoe deficiency attenuated tumor growth and was accompanied by reduced vascularization and increased CD8-positive cell infiltration, whereas tumor-cell Apoe knockdown did not significantly affect tumor volume or tumor weight in immunodeficient mice. In the Nras-Myc liver tumor model, Apoe deficiency reduced tumor burden and proliferative activity and was accompanied by a shift in tumor macrophage and monocyte programs away from lipid-processing and phagolysosomal activity toward inflammatory chemokine-associated activity. In lipid-loaded bone marrow-derived macrophages, Apoe deficiency increased neutral lipid accumulation.ConclusionThis study links APOE-associated macrophage lipid handling to inferred ligand–receptor communication and host Apoe-linked microenvironmental remodeling in HCC. These findings highlight an APOE-linked myeloid lipid-handling program as a potential component of immune microenvironmental remodeling during HCC development.

Abstract Background Activation of the host immune system is a promising strategy for treating hepatitis B virus (HBV). The stimulator of interferon genes (STING) signaling pathway acts as an innate immune sensor for viral DNA. We aimed to determine whether STING activation can inhibit HBV replication in the liver. Methods We evaluated STING expression in peripheral blood mononuclear cells and liver biopsy specimens from people with chronic hepatitis B (CHB). We examined the effect of the STING agonist 5,6-dimethylxanthenone-4-acetic acid (DMXAA) on HBV replication and the immune response in a mouse model with persistent HBV replication (rAAV8-HBV1.3). Additionally, we depleted macrophages in vivo to assess their role in DMXAA's antiviral effects. Results Stimulator of interferon genes expression in people with CHB was higher in the immune-active phase than in the inactive phase or uninfected healthy controls, suggesting an association between elevated STING expression and immune activation. In a mouse model with persistent HBV replication, DMXAA reduced serum HBV DNA in a dose-dependent manner, whereas repeated administration reduced serum HBV DNA, hepatitis B surface antigen, and hepatitis B e antigen levels and enhanced hepatic immune responses. Transcriptome sequencing of liver tissues revealed 856 differentially expressed genes involved in immune-related signaling pathways. In hepatic macrophage-depleted mice, the reductions in HBV virological markers following DMXAA treatment were less pronounced. Conclusions Stimulator of interferon genes pathway activation enhances the immune response against HBV by upregulating the expression of immune-related genes. Hepatic macrophages play an important role in STING-mediated inhibition of HBV replication.