Hepatology Communications最新文献

Background: As the role of hepatic progenitor cells (HPCs) in constituting ductular reactions in pathogenesis remains ambiguous, we aimed to establish the in vivo cause-and-effect relationship between HPCs and chronic liver disease progression. We previously demonstrated that peritumoral ductules are associated with angiogenesis in liver tumors, and forkhead box L1 (Foxl1)-expressing murine HPCs secrete angiogenic factors in vitro. Therefore, we hypothesized that HPCs are capable of remodeling the portal vascular microenvironment and regulating overall liver disease progression, and this function of HPCs is dependent on recombination signal binding protein for immunoglobulin kappa J region (RBPJ), a key effector of the Notch signaling pathway.

Methods: We generated HPC-specific Rbpj conditional knockout mice (CKO) using Foxl1-Cre, treated them with the DDC diet to induce chronic liver disease, and performed serum biochemistry, gene expression analysis, and immunostaining analysis.

Results: CKO mice exhibited a significant reduction in serum levels of liver injury markers, ductular reactions, vascular and fibrotic areas, and hepatic expression of fibrosis and inflammation markers compared to control mice (WT). Single-nucleus RNA sequencing comparing CKO and WT livers detected transcriptome changes across multiple cell types, including endothelial cells, HSCs, and cholangiocytes. Expression of several reactive cholangiocyte markers, including vascular cell adhesion molecule 1 (VCAM1), in HPCs was significantly downregulated in response to anti-Rbpj shRNAs in vitro. Immunofluorescence analysis indicated that the percentage of VCAM1+ cells was reduced in both HPC and cholangiocyte populations in CKO compared to WT in vivo.

Conclusions: Our findings reveal Rbpj-dependent expression of reactive cholangiocyte markers in HPCs and demonstrate that Rbpj deletion in HPCs attenuates not only endothelial responses but also liver injury, fibrosis, and VCAM1 expression in cholangiocytes, highlighting the crucial role of HPCs in pathogenic progression.

Background: Acute-on-chronic liver failure (ACLF), which develops in patients with acutely decompensated cirrhosis, is characterized by multiple extrahepatic organ failures leading to high short-term mortality. Although major advances in the understanding of ACLF have been accomplished in the last years, the understanding of driving mechanisms underlying ACLF is hindered by the lack of proper animal models that faithfully reproduce both the systemic hyperinflammatory response and the full spectra of extrahepatic organ failures present in this condition.

Methods: ACLF was induced by acute induction of polymicrobial peritonitis secondary to the ligation and puncture of the cecum (CLP) in mice with chronic carbon tetrachloride (CCl4)-induced cirrhosis. The study included three groups: CCl4+CLP (n=10) mice with cirrhosis which underwent CLP surgery; CCl4+sham mice (n=10) and control mice (n=10).

Results: As compared to CCl4+sham, CCl4+CLP mice had higher short-term mortality and exhibited more severe hypoalbuminemia and hyperbilirubinemia, significantly higher AST and GGT levels and higher liver inflammatory burden. CCl4+CLP mice also showed increased serum creatinine and BUN levels and up-regulated expression of Kim-1, Il-6 and Tnf in the kidney, lower oxygen saturation (SpO2), higher serum renin concentration, higher international normalized ratio (INR) and worse neurological behavior test scores than CCl4+sham and control mice. In addition, CCl4+CLP mice showed widespread bacterial tissue colonization and exhibited increased serum cytokine levels, which correlated with the intensity of organ impairments.

Conclusion: The CCl4+CLP model reproduces the full spectra of extrahepatic organ impairments present in patients with ACLF and represents an optimized murine model to experimentally explore the pathophysiology of this disease as well as new therapeutic approaches.

Background: Metabolic-associated steatotic liver disease (MASLD), caused by insulin resistance and the metabolic syndrome, may result in progressive liver fibrosis. Animal studies suggest that dietary content modulates liver fibrosis progression. Our aim was to identify dietary components and food-related behaviors that may be associated with fibrosis progression and liver-related outcomes in a well-characterized human MASLD cohort.

Methods: Patients with MASLD who had completed a detailed Lifestyle Survey, including a semiquantitative Food Frequency Questionnaire in the Veterans Health Administration Million Veteran Program, were included. The primary outcome was liver fibrosis progression using the Fibrosis-4 slope; the secondary outcome was time to cirrhosis by ICD9/10 codes. Key baseline covariates included: race/ethnicity, body mass index, diabetes mellitus, AUDIT-C score, and baseline Fibrosis-4 score. Using bootstrapped Elastic Net regression in R, self-reported food intake and scaled nutrient variables of interest associated with the outcomes were identified and then validated using multivariable Generalized Linear Model and Cox models.

Results: A total of 84,024 individuals with MASLD with nutritional data were included in this study. Median age at MASLD diagnosis was 56 years (IQR 49-63). Frequency of consumption of coffee, tea, vegetables (broccoli, spinach/collard greens), legumes, nuts, modest alcohol, white meat, rice/pasta, dairy, and intakes of specific nutrients including nitrate/vitamin K, caffeine, betaine, amino acids, and beta carotene were associated with reduced fibrosis progression. Consumption of white bread, cookies, breakfast cereals, and specific nutrients such as iron (non-heme), B vitamins, and flavanones were all significantly associated with increased fibrosis progression in MASLD (p<0.05).

Conclusions: Dietary choices such as intake of processed foods, high-fructose foods, and refined carbohydrates may be associated with MASLD progression, while intake of vegetables, nuts, whole grains, and caffeine may be protective.

Background: HBV, which contributes to liver fibrosis and HCC, is associated with autophagy and epithelial-mesenchymal transition (EMT). However, the relationship between HBV infection, autophagy, and EMT remains unknown.

Methods: In this study, we used HBV cell lines (HepAD38, Huh7.5.1-NTCP) to evaluate this relationship.

Results: HBV replication or infection promoted autophagy, EMT, and liver fibrogenesis. HBV-induced EMT/fibrogenesis was dependent on autophagy. HBV X and HBV core (C) were each involved during this process. TGF-β1 partially participated in HBV-induced autophagy/EMT/fibrosis. c-Jun N-terminal kinase (JNK) expression was increased in HepAD38 cells, HBV-X (HBV-C)-transfected Huh7.5.1 cells, and HBV-infected Huh7.5.1-NTCP cells. JNK silencing attenuated HBV-induced autophagy and EMT/fibrosis signaling as well. Moreover, overexpression of HBV-X(C) and augmentation of autophagy were shown to promote pJNK expression. Finally, we confirmed an HBV-JNK-autophagy-EMT/fibrosis signaling pathway in a primary human hepatocyte model.

Conclusions: In conclusion, HBV induces autophagy through JNK, thus triggering downstream EMT and fibrogenesis signaling pathways. These findings suggest that JNKs could be potential targets for the treatment of HBV-related liver diseases.

Background: Racial and ethnic minority populations are disproportionately impacted by HCC due to more advanced tumor burden and underuse of treatments. We explored racial and ethnic differences in medical mistrust, barriers to treatment, and health literacy among patients with HCC.

Methods: We conducted a multicenter survey among patients with newly diagnosed HCC between September 2018 and July 2023 at 4 large U.S. health systems. The survey assessed medical mistrust [Group-Based Medical Mistrust Scale (GBMMS)], health literacy (CHEW Assessment of Health Literacy), and barriers to HCC treatment. We performed multivariable logistic regression to evaluate associations between race and ethnicity and survey measures.

Results: Of 1245 eligible patients, 833 (66.9%) completed the survey (45.9% Hispanic, 35.9% White, and 14.2% Black). A higher proportion of Black and Hispanic patients had high medical mistrust than White patients (14.2% and 3.3% vs. 0.7%, respectively; p<0.001). In multivariable analysis, Black race (OR: 19.2, 95% CI: 4.2-87.7) but not Hispanic ethnicity (OR: 3.72, 95% CI: 0.80-17.2) was significantly associated with high mistrust. Compared to White patients, Black and Hispanic patients both reported greater barriers to HCC treatment, with the most common barriers being concerns about pain (41.6%), financial burden (37.6%), and time commitment (31.1%). Limited health literacy was reported by 38.1% of patients (46.8% Hispanic, 41.0% Black, 26.2% White; p<0.001).

Conclusions: Medical mistrust, barriers to treatment, and limited health literacy are prevalent among Black and Hispanic patients with HCC. Understanding the interplay between race, ethnicity, and these factors is essential to address HCC disparities.

Background: HCC is a significant health concern. CTNNB1 mutations are implicated in HCC progression and resistance to transarterial chemoembolization (TACE), potentially through the ITGB1/PI3K/AKT pathway.

Methods: HCC was induced in mice using diethylnitrosamine, and TACE-resistant models were established. Tumor tissue analysis, single-cell and whole-exome sequencing identified gene mutations and cellular interactions. CRISPR/Cas9 was used to generate HCC cells with CTNNB1 mutations, and functional assays evaluated their proliferation, migration, and invasion. Cocultivation with HUVEC cells and animal models assessed angiogenesis and tumorigenesis.

Results: The study successfully established a TACE-resistant mouse model, identifying mesenchymal cell alterations and enhanced cellular communication in resistant mice. Signaling pathways like SPP1 were implicated in epithelial-mesenchymal transition. Analysis revealed a CTNNB1 (c.890T>C) mutation in TACE-resistant patients, with subsequent experiments confirming enhanced proliferation, migration, and epithelial-mesenchymal transition in CTNNB1 mutant HCC cells. Cocultivation studies with HUVEC cells indicated a pro-angiogenic effect of CTNNB1 mutant HCC cells, mediated by the ITGB1 pathway. Animal experiments demonstrated tumorigenic properties of CTNNB1 mutant cells, further validated by histopathological and immunohistochemical analyses.

Conclusions: CTNNB1 mutations elevate ITGB1, activate PI3K/AKT, induce epithelial-mesenchymal transition, enhancing proliferation, migration, and angiogenesis, contributing to TACE resistance, suggesting novel therapeutic targets in HCC through signaling pathway interventions.

Background: To investigate the role of nucleoside diphosphate kinase 2 (NME2) in HCC progression, assessing its therapeutic potential.

Methods: Utilizing transcriptome sequencing data from The Cancer Genome Atlas (TCGA) and immunohistochemical staining of tissue microarrays, we analyzed NME2 expression in HCC tumor tissues. The effects of NME2 on HCC cell proliferation and autophagy flux were assessed through knockdown and overexpression experiments. Additionally, the relationship between NME2 and 4EBP1 phosphorylation was explored through specific site mutation analysis.

Results: NME2 overexpression in HCC correlated with poor prognosis. NME2 knockdown significantly hindered HCC cell proliferation and induced autophagy flux. Notably, NME2 modulates 4EBP1 phosphorylation (Thr37/46) independently of mTOR, unveiling a novel axis in HCC pathogenesis. Additionally, NME2 modulates eukaryotic translation initiation factor 4F (eIF4F) complex formation and autophagy flux.

Conclusions: NME2 plays a crucial role in HCC development by modulating 4EBP1 phosphorylation and autophagy through an mTOR-independent pathway. Our research underscores NME2's significance as a potential therapeutic target in HCC, meriting further exploration of its underlying mechanisms and clinical applicability.

Background: For infants with biliary atresia, the only treatment that can establish bile flow and delay need for liver transplant is the Kasai portoenterostomy (KP). Unfortunately, the KP has variable success. In this study, we hypothesized that intravenous N-acetylcysteine (IV NAC) treatment following KP would improve bile flow.

Methods: This was a phase 2 study following the two-stage "minimax" trial design. Participants received IV NAC (150 mg/kg/day) for 7 days after KP, and the primary endpoint was achieving total serum bile acids (TSBA) ≤10 μmol/L within 24 weeks of KP. Secondary endpoints were clinical markers and the occurrence of sentinel events.

Results: There were 12 participants in stage 1 who received treatment, with none achieving TSBAs ≤10 μmol/L within 24 weeks of KP. As a result, no participants were enrolled in stage 2. There were 32 adverse events in 11 participants, including 5 serious adverse events which were considered part of the participants' natural clinical course and not directly attributable to NAC treatment. Analyses of secondary outcomes demonstrated no difference in clinical markers or occurrence of sentinel events between study participants and matched historical controls.

Conclusions: This study demonstrates how the two-stage "minimax" trial design can be used to efficiently evaluate potential therapies for BA. Although the primary endpoint was not met, NAC therapy was generally well-tolerated. NAC therapy may prove efficacious in future trials with (i) a less stringent primary endpoint and/or (ii) a longer course of treatment (NCT03499249).

Background: Predictive biomarkers are needed to identify individuals with metabolic dysfunction-associated steatotic liver disease (MASLD), at high risk for HCC. Our study aimed to determine whether the detection of circulating bacterial DNA could be associated with HCC development in MASLD patients with liver cirrhosis.

Methods: We developed a multicenter prospective cohort of patients with cirrhosis undergoing surveillance for HCC by contrast-enhanced magnetic resonance imaging. In a nested cohort study, we performed 16S rRNA sequencing of cell-free DNA extracted from 343 longitudinal plasma samples collected from 151 MASLD patients with cirrhosis. Among the 151 patients, 25 developed HCC during follow-up.

Results: Following in silico decontamination approaches, variations in circulating bacterial DNA profiles were significantly associated with HCC development, cirrhosis severity, and male gender. Many of the identified taxa were well-known human-associated bacteria. Patients who developed HCC during follow-up showed an enrichment in Tsukamurella and Bacteroides, and depletion of Natronomonas. Associations with HCC development remained for the identified bacteria, after adjusting for cirrhosis severity and male gender. Acidobacteriota, Tsukamurella, and Staphylococcaceae showed markedly improved prediction of HCC within 12 months prior to diagnosis [Acidobacteriota: AOR=2.87 (1.36-6.04), p=0.006; Tsukamurella: AOR=2.80 (1.34-5.85), p=0.006; Staphylococcaceae: AOR=2.52 (1.19-5.36), p=0.016]. Circulating bacterial DNA profiles associated with male gender and cirrhosis severity were different from those observed for HCC and showed considerable overlap in significant taxa. These included enrichment of the lineage Gammaproteobacteria, Enterobacterales, and Rheinheimera.

Conclusions: The identified circulating bacterial DNA signatures may have utility in personalized approaches to HCC surveillance in MASLD patients.