Hepatology Communications最新文献

Background: Glutamine (Gln) is a critical amino acid for energy expenditure. It participates in extracellular matrix (ECM) formation and circulates in the hepatic parenchyma in a spatial-oriented manner. Posthepatectomy liver mass recovery poses a regenerative challenge. However, little is known about the role of Gln in liver regeneration, notably the spatial orientation in the remodeling process. This study aimed to elucidate Gln-potentiated liver regeneration and ECM remodeling after mass loss.

Methods: We studied the regenerative process in hepatectomized mice supplemented with Gln. Second harmonic generation/two-photon excitation fluorescence microscopy, an artificial intelligence-assisted structure-based imaging, was used to demonstrate the spatial-oriented process in a hepatic acinus.

Results: Gln promotes liver mass regrowth through the cell cycle, Gln metabolism, and adipogenesis pathways after hepatectomy. Ornithine transaminase, one of the upregulated enzymes, showed temporal, spatial, and functional correspondence with the regeneration process. Second harmonic generation/two-photon excitation fluorescence microscopy highlighted transient hepatic steatosis and ECM collagen synthesis, predominantly in the portal tract instead of the central vein area. Structural remodeling was also observed in the portal tract area.

Conclusions: Gln promotes liver regeneration through cellular proliferation and metabolic reprogramming after hepatectomy. Using structure-based imaging, we found that Gln potentiated hepatic steatosis and ECM collagen deposition predominantly in the portal tract area. These results highlighted the spatial orientation and mechanistic implications of Gln in liver regeneration.

Background: Postoperative liver failure due to insufficient liver cell quantity and function remains a major cause of mortality following surgery. Hence, additional investigation and elucidation are required concerning suitable surgeries for promoting in vivo regeneration.

Methods: We established the portal vein ligation (PVL) and associated liver partition and portal vein ligation for staged hepatectomy (ALPPS) mouse models to compare their in vivo regeneration capacity. Then, RNA-seq and microRNA-seq were conducted on the livers from both mouse models. Weighted gene co-expression network analysis algorithm was leveraged to identify crucial gene modules. ScRNA-seq analysis was used to understand the distinctions between Signature30high hepatocytes and Signature30low hepatocytes. Moreover, in vivo, validation was performed in fumarylacetoacetate hydrolase knockout mice with gene editing using the CRISPR-cas9 system. A dual luciferase report system was carried out to further identify the regulatory mechanisms.

Results: RNA-seq analysis revealed that ALPPS could better promote cell proliferation compared to the sham and portal vein ligation models. Moreover, a Plk1-related 30-gene signature was identified to predict the cell state. ScRNA-seq analysis confirmed that signature30high hepatocytes had stronger proliferative ability than signature30low hepatocytes. Using microRNA-seq analysis, we identified 53 microRNAs that were time-dependently reduced after ALPPS. Finally, miR-30a-3p might be able to regulate the expression of Plk1, contributing to the liver regeneration of ALPPS.

Conclusions: ALPPS could successfully promote liver regeneration by activating hepatocytes into a proliferative state. Moreover, a Plk1-related 30-gene signature was identified to predict the cell state of hepatocytes. miR-30a-3p might be able to regulate the expression of Plk1, contributing to the liver regeneration of ALPPS.

Background: The incidence of alcohol-associated hepatitis (AH) is rising in women of reproductive age. While the adverse effects of alcohol on pregnancy are well documented, there is limited data on pregnancy in women with a history of AH.

Methods: This study was completed by using the TriNetX Research Network. The primary objectives were to evaluate the incidence of pregnancy and related complications in pregnancies following an episode of AH (AH pregnancies) compared to pregnancies in healthy patients (control pregnancies). The secondary objective was to assess long-term liver-related complications and mortality in women with AH who experienced a pregnancy compared to no pregnancy. Propensity score matching was used for comparative analyses to balance cohorts by age, race, ethnicity, prior delivery, and obesity status.

Results: The incidence of pregnancy was significantly lower in women with AH compared to controls (26 vs. 54 cases per 1000 person-years, p<0.001). AH pregnancies were associated with higher odds of spontaneous abortion (OR 2.0, 95% CI: 1.2 to 3.3, p=0.011), pre-eclampsia (OR 1.9, 95% CI: 1.1 to 3.0, p=0.002), peri-partum hemorrhage (OR 2.7, 95% CI: 1.3 to 5.6, p=0.007) and perinatal psychiatric disorders (OR 3.2, 95% CI: 1.6 to 6.2, p=0.001). The incidence of cirrhosis and hepatic decompensation were similar between women with AH who experienced a pregnancy compared to no pregnancy, but Kaplan Meier analysis revealed a significantly faster time to event in the no-pregnancy group.

Conclusions: Pregnancies following AH diagnosis were associated with adverse pregnancy outcomes. Pregnancy after AH does not reduce the overall risk of developing advanced liver disease but may delay disease progression. These findings highlight the importance of tailored reproductive counseling and support for this population.

Background: Liver progenitor cells (LPCs) with bipotential differentiation capacities are essential for restoring liver homeostasis and hepatocyte population after damage. However, the low proportion and shared markers with epithelial cells make studying LPC heterogeneity difficult, especially in humans. To address this gap, we explored over 259,400 human liver single cells across 4 conditions (fetal, healthy, cirrhotic, and HCC-affected livers).

Methods: Human liver tissue samples were analyzed using spatial transcriptomics sequencing technologies to describe the heterogeneity of LPCs. Liver tissue was characterized by LPC heterogeneity at single-cell resolution by employing cellular modules, differentiation trajectories, and gene co-expression patterns.

Results: We annotated and identified 1 LPC cluster, 3 LPC subpopulations, and 4 distinct cellular modules, indicating the heterogeneity within LPC and the diversity between LPCs and epithelial cells. LPCs showed spatial colocalization with cholangiocytes and comprised a small proportion (2.95±1.91%) within the merged epithelial cells and LPC populations, exhibiting marked differences in marker expression patterns compared to those in mice. LPCs exhibited distinct cellular states in functional restoration, activation, proliferation, and cell transition. Additionally, the gene co-expression network of LPCs exhibited 3 unique modules, reflecting the distinct connectivity of genes encoding apolipoproteins and heat shock proteins in the gene co-expression network modules.

Conclusions: Our study provides valuable insights into the multifaceted heterogeneity of human LPCs. Future studies focusing on spatial gene expression dynamics will contribute to our understanding of the spatial arrangement of liver regeneration.

Background: Acetaminophen-induced acute liver injury (AILI) is one of the common causes of abrupt liver failure in numerous nations. Several previous studies revealed that tiliroside, a glycoside flavonoid, exerts neuroprotective and renal protective effects. However, whether it has hepatoprotective effects is not known. The objective of this research is to examine whether tiliroside can protect against AILI.

Methods: AILI mouse and cell models were performed to evaluate the protective effects of tiliroside. Molecular docking, cellular thermal shift assay, immunoprecipitation, and RNA-seq were performed to analyze the possible mechanisms of tiliroside.

Results: In vivo, tiliroside attenuated AILI in mice significantly, as evidenced by lower ALT and AST levels. Molecular docking, cellular thermal shift assay, and RNA-seq analysis revealed that tiliroside promoted the activation of nuclear factor erythroid 2-related factor 2 (NRF2) and the expression of its downstream genes through disruption of the NRF2-KEAP1 protein-protein interaction to inhibit KEAP1-mediated ubiquitination and degradation of NRF2, thereby inhibiting oxidative stress in the livers of AILI mice. Furthermore, hepatocyte-specific knockout of NRF2 greatly attenuated the hepatic-protective effects of tiliroside in mice. In vitro, tiliroside protected against acetaminophen-induced oxidative stress on cultured hepatocytes through activation of NRF2. In addition, NRF2 knockout markedly blunted the protection effects of tiliroside, suggesting that NRF2 mediates the hepatic-protective effects of tiliroside.

Conclusions: Our study demonstrated that tiliroside could protect against AILI by activating the KEAP1/NRF2 pathway, which primarily inhibits the processing of oxidative stress and cell death. Our results suggest that tiliroside could serve as a potential agent for the clinical treatment of AILI.

Background: Immune checkpoint inhibitors are effective treatments for HCC; however, their therapeutic efficacy is often limited by the development of drug resistance. Therefore, investigating new combination therapeutics involving immune checkpoint inhibitors is critical to improving patient prognosis. In this study, we investigated the therapeutic effect of cordycepin (COR) in HCC and its synergistic effect with anti-programmed cell death ligand 1 (anti-PD-L1) immunotherapy.

Methods: We selected 2 HCC cell lines to investigate the effects of COR on HCC growth using in vivo and in vitro experiments. We performed RNA sequencing of the MHCC97H cell line treated with or without COR to understand the underlying mechanism and identify the key regulatory genes. Through in vivo and in vitro experiments on gene knockdown cells, we identified thioredoxin-interacting protein as a key molecule involved in the role of COR. Next, we used mouse subcutaneous and orthotopic tumor models to evaluate the therapeutic effects of COR, atezolizumab (a programmed death-ligand 1 [PD-L1] inhibitor), or their combination. Multiple immunofluorescence staining revealed that the combination of atezolizumab and COR therapy greatly increased the number of tumor-infiltrating CD8+ T cells and PD-L1 expression in HCC compared to monotherapy.

Results: Our study revealed that COR significantly inhibited HCC growth both in vitro and in vivo. Mechanistically, we showed that COR induces endoplasmic reticulum stress, which upregulates thioredoxin-interacting protein expression and leads to HCC cell pyroptosis. In addition, the combination treatment with COR and PD-L1 inhibitors profoundly inhibited HCC.

Conclusions: Overall, our study successfully established a combined therapeutic strategy using COR and PD-L1 inhibitors. This strategy has significant synergistic effects on cancer cells, highlighting its importance in cancer therapy.

Background: Transarterial chemoembolization (TACE) is the primary treatment modality for advanced HCC, yet its efficacy assessment and prognosis prediction largely depend on imaging and serological markers that possess inherent limitations in terms of real-time capability, sensitivity, and specificity. Here, we explored whether multiple features of cell-free mitochondrial DNA (cf-mtDNA), including copy number, mutations, and fragmentomics, could be used to predict the response and prognosis of patients with HCC undergoing TACE treatment.

Methods: A total of 60 plasma cell-free DNA samples were collected from 30 patients with HCC before and after the first TACE treatment and then subjected to capture-based mtDNA sequencing and whole-genome sequencing.

Results: Comprehensive analyses revealed a clear association between cf-mtDNA multiple features and tumor characteristics. Based on cf-mtDNA multiple features, we also developed HCC death and progression risk prediction models. Kaplan-Meier curve analyses revealed that the high-death risk or high-progression-risk group had significantly shorter median overall survival (OS) and progression-free survival than the low-death risk or low-progression-risk group (all p<0.05). Moreover, the change in cf-mtDNA multiple features before and after TACE treatment exhibited an exceptional ability to predict the risk of death and progression in patients with HCC (log-rank test, all p<0.01; HRs: 0.36 and 0.33, respectively). Furthermore, we observed the consistency of change between the cf-mtDNA multiple features and copy number variant burden before and after TACE treatment in 40.00% (12/30) patients with HCC.

Conclusions: Altogether, we developed a novel strategy based on profiling of cf-mtDNA multiple features for prognosis prediction and efficacy evaluation in patients with HCC undergoing TACE treatment.

Background: Intrahepatic cholangiocarcinoma (ICC) is a poor prognosis of malignant cancer with high lymph node metastasis and resistance to systemic therapies. Recent studies suggested that the involvement of IL-8 could promote ICC metastasis through epithelial-mesenchymal transition while the ICC-ALDH1A1high subtype is clarified by multi-omics study. The correlation between ALDH1A1 and IL-8 in ICC remains elusive. This study aims to further explore the roles and regulatory mechanisms of ALDH1A1 and IL-8 in ICC.

Methods: We analyzed IL-8 and ALDH1A1 expression in ICC patients and cells. CXCR2 inhibitor (SB225002) was applied to inhibit the function of IL-8, and JSH-23 was applied to inhibit the NF-κB signaling pathway. We examined the effects of IL-8 inhibition on NF-κB, ALDH1A1 expression, and cell growth, migration, invasion, and stemness. Moreover, we examined the effects of ALDH1A1 on NF-κB, IL-8 expression, and cell growth, migration, invasion, and stemness. The effects of IL-8 and ALDH1A1 on tumor growth and NF-κB expression were validated using subcutaneous tumors in nude mice.

Results: IL-8-derived tumor cells could promote ICC progression. The high expression of IL-8 in serum was associated with lymph node metastasis. IL-8 could upregulate ALDH1A1 expression by activating the NF-κB signaling pathway, promoting tumor progression. Upregulation of ALDH1A1 could activate NF-κB to promote IL-8 secretion, forming a positive feedback loop to promote tumor invasiveness and cell stemness in ICC.

Conclusions: IL-8-derived tumor cells could upregulate ALDH1A1 expression by activating the NF-κB signaling pathway, promoting tumor progression. Upregulation of ALDH1A1 could activate NF-κB to promote IL-8 secretion, forming a positive feedback loop to promote tumor invasiveness and cell stemness in ICC.

Background: In the nucleos(t)ide analog (NA)-control arm of the REEF-2 study (NCT04129554), virologic relapse (confirmed increase in HBV DNA >2000 IU/mL) and biochemical flare (ALT increases ≥3× upper limit of normal) were frequently observed after stopping NA treatment. We characterized the posttreatment virologic relapses and biochemical flares and assessed their association with end-of-treatment (EOT) HBV serum markers.

Methods: In REEF-2, a randomized-controlled study, virologically suppressed HBeAg-negative patients stopped treatment at week 48, followed by 48 weeks of follow-up. EOT HBV RNA, hepatitis B core-related antigen, and quantitative anti-hepatitis B core (HBc) IgG levels were assessed in 41/45 NA-control arm patients; their association with off-treatment response was evaluated.

Results: A similar proportion of patients with EOT HBV RNA or hepatitis B core-related antigen detectable and target not detectable had virologic relapse or ALT flares (p>0.05). A higher frequency of severe virologic relapse (peak HBV DNA >100,000 IU/mL) and/or severe biochemical flares (peak ALT ≥10× upper limit of normal) was observed in patients with EOT detectable hepatitis B core-related antigen levels, HBsAg <1000 IU/mL, and/or anti-HBc IgG titers <300 IU/mL, respectively (p<0.05). None of the 11 patients with EOT anti-HBc titers ≥300 IU/mL had severe virologic or biochemical flare off treatment (100% positive predictive value and 48% negative predictive value).

Conclusions: In this prospective study of patients who stopped NA, anti-HBc levels ≥300 IU/mL were associated with a low risk of developing virologic relapse and severe biochemical flares. Future research should confirm a potential protective effect of high anti-HBc IgG levels.

Background: Patients with HCC face numerous barriers to curative therapies, particularly Black patients and those impacted by adverse social determinants of health (SDOH). This study aimed to identify patient-reported barriers and facilitators to curative therapies, to inform interventions that improve equitable access to care.

Methods: We conducted 2 qualitative sessions with Black participants and participants experiencing adverse SDOH with HCC referred for liver transplant (LT) or resection. We also conducted one-on-one interviews with participants from sessions that underwent LT (n=2). Human-centered design methods, including journey mapping and group ideation, were used to identify challenges and solutions at various stages in the care pathway. Data were analyzed to identify key themes and to compare the experiences of Black patients with those experiencing adverse SDOH.

Results: Both groups faced significant barriers, particularly related to information overload, communication gaps with health care providers, and the complexity of navigating the LT pathway. However, Black patients reported additional challenges related to the psychological burden of the diagnosis and distrust in the health care system, while those with adverse SDOH frequently cited financial instability, lack of social support, and challenges in coordinating care between multiple health systems. Despite these differences, common facilitators included compassionate health care teams and strong personal support networks. Both groups suggested solutions such as improvements in education timing and delivery, better communication pathways, and peer support groups to improve preparedness for treatment and recovery.

Conclusions: While Black patients and those with adverse SDOH experience unique barriers, common threads-such as information gaps and desire for peer support suggest shared opportunities for interventions.