Objective:To investigate the role and underlying mechanism of Six-Transmembrane Epithelial Antigen of the Prostate 1 (STEAP1) in Hepatocellular carcinoma (HCC) progression, and to validate its potential as a therapeutic target. Materials and Methods:In this study, STEAP1-knockdown models were used to evaluate cell proliferation, invasion and migration. RNA sequencing (RNA-seq) were performed to explore downstream mechanisms. Subcutaneous Xenograft Model in nude mice was used to investigate the role of STEAP1 in the proliferation of HCC in vivo. Results:STEAP1 knockdown significantly inhibited HCC cell proliferation, invasion, and migration, downregulated the expression of EMT-related proteins, and suppressed activation of the Wnt signaling pathway. In vivo, STEAP1 silencing effectively reduced the growth of subcutaneous tumors in nude mice. Conclusion:STEAP1 promotes HCC growth and metastasis by activating the Wnt/β-catenin signaling pathway, and may serve as a promising therapeutic target for HCC.

INTRODUCTION:The tumor microenvironment (TME) promotes immune evasion by fostering regulatory immune programs. Although CD8+ regulatory T cells have been described, their identity, upstream drivers and impact in tumors remain incompletely defined. OBJECTIVES:To define the phenotype, regulation determinants, and functional of CD86high CD8+ T cells in tumor immunity. METHODS:Murine tumor models were used to isolate tumor-infiltrating lymphocytes (TILs) for flow cytometry (FCM), functional co-cultures, and RNA-seq. Upstream cues were probed through cytokine stimulation and transcription factor analyses. Dendritic cells (DCs) conditioning was tested with bone marrow derived DCs and OT-I/OT-II antigen-specific systems. The in vivo function of CD86 on CD8+ T cells was evaluated using conditional Cd86 knockout in CD8+ T cells mouse model. RESULTS:CD86 was markedly upregulated on tumor-infiltrating CD8+ T cells, defining a CD86high subset that accumulated in tumors. Transcriptomic profiling revealed enrichment of immunoregulatory and terminal-exhaustion programs while preserving effector modules. CD86high cells upregulated Il12rb1 and IRF5; exogenous IL-12 increased the frequency and intensity of CD86 expression showing dose and time dependent effects, whereas IRF5 inhibition curtailed this induction, establishing an IL-12-IRF5 axis controlling CD86. Functionally, CD86high CD8+ T cells selectively suppressed antigen-specific OT-I/OT-II responses, reducing IFN-γ, IL-2, and Ki-67, and reprogrammed DCs toward a regulatory phenotype characterized by increased IDO, IL-10, CTLA-4, and CD39. Genetic ablation of Cd86 in CD8+ T cells reduced tumor growth, diminished CD39 and PD-1 on TILs, enhanced IFN-γ and granzyme B, and remodeled the myeloid compartment toward immunostimulatory DCs with elevated CD40/CD80/CD86 and reduced IDO/IL-10/CTLA-4/CD39. CONCLUSIONS:CD86high CD8+ T cells constitute a distinct immunoregulatory subset in cancer. Their differentiation is driven by an IL-12-IRF5 program, and their crosstalk with DCs via CD86/CTLA-4 engagement promotes tolerogenic remodeling of the TME. Targeting CD86 on CD8+ T cells may disrupt this suppressive circuit and potentiate antitumor immunity.

Despite recent progresses in microbiome and infection, the role of multi-kingdom gut microbiome in kidney transplantation (KT) infection remains unexplored. Here we performed a longitudinal and integrative multi-omics analysis of the gut microbiome, fecal metabolome and plasma metabolome in 169 KT recipients across 5 different transplantation centers, comprising discovery and validation cohorts. We observed KT-specific four kingdom microbiome dysbiosis, including bacteria, fungi, archaea and viruses, with the most pronounced shifts in bacterial and fungal communities. Furthermore, we identified 6 infection-associated co-abundance groups (CAGs) composed of 23 bacterial and 3 fungal species, highlighting extensive bacterial-fungal interactions. Interestingly, infection-associated fecal metabolomic pattern F1, enriched in N-acetylputrescine and hydroxyproline, was positively correlated with Enterococcus-, Citrobacter- and Lactococcus-dominated CAGs, as well as the plasma metabolite signature, represented by phenylacetyl-l-glutamine, indoxyl sulfate and leukotriene. In contrast, cholesterol sulfate and menadione in plasma were aligned with fecal indoleacetic acid and stachyose, a metabolic signature more characteristic of non-infected recipients. Finally, the combinatorial biomarkers of fungal and bacterial species achieved powerful diagnosis ability of KT infection in an independent validation cohort (area under the receiver operating characteristic curve (AUROC) = 0.80) with the fecal metabolites achieving high accuracy (AUROC = 0.83). Collectively, our findings not only uncovered the postoperative infection-specific multi-kingdom microbial network dynamics, but also revealed the microbial and its metabolic biomarkers with powerful diagnostic ability for postoperative infection in kidney transplantation.

Liver transplantation (LT) remains the only effective treatment for end-stage liver disease. Autophagy critically regulates liver ischemia–reperfusion (I/R) injury. Transmembrane 9 superfamily member 1 (TM9SF1) is a transmembrane protein associated with the elevation of autophagy. However, the role and mechanism of this protein in liver I/R injury have not been explored. We observed upregulated TM9SF1 expression in liver I/R mice models and AML12 cells subjected to hypoxia-reoxygenation (H/R). Using TM9SF1 adeno-associated virus (AAV) to generate overexpression and knockdown (KD) mice with liver I/R injury, we found that TM9SF1-overexpressing mice exhibited exacerbated liver damage, inflammation, and autophagy, whereas KD-TM9SF1 mice showed opposite results. Mechanistically, we found that TM9SF1 and Annexin A2 (ANXA2) interacted and jointly promoted the expression of autophagy levels during liver I/R injury. Virtual screening of FDA-approved compounds identified lomitapide as an inhibitor that selectively suppresses TM9SF1 expression, thereby attenuating I/R injury. In general, our findings indicated that TM9SF1 and ANXA2 interact with each other, promoting autophagy levels through activating the mitogen-activated protein kinase (MAPK) pathway, thereby aggravating liver I/R injury. Targeting TM9SF1-ANXA2 may be a potential therapeutic strategy.

BACKGROUND:Endoplasmic reticulum membrane protein complex subunit 3 (EMC3) plays a critical role in protein translocation and processing and has been implicated in multiple cancer types. However, its specific function and clinical relevance in hepatocellular carcinoma (HCC) remain poorly understood. This study aimed to investigate the molecular role and prognostic significance of EMC3 in HCC progression. METHODS:Public datasets and clinical samples from HCC patients were analyzed to assess EMC3 expression and its association with prognosis. Genomic and immunomic profiles were evaluated to explore EMC3-related genomic instability and its regulation of the tumor immune microenvironment (TIME). Functional assays, including clone formation, CCK-8, EdU proliferation, wound healing, Transwell migration and invasion, and in vivo xenograft and lung metastasis models, were performed to examine the effects of EMC3 on HCC cell proliferation and epithelial-mesenchymal transition (EMT). Mechanistic studies involved RNA sequencing, pathway enrichment analysis, and rescue experiments using the PI3K inhibitor LY294002. RESULTS:EMC3 was significantly overexpressed in HCC tissues and correlated with poor patient survival. Genomic analyses revealed EMC3-associated instability, and immunomic profiling indicated its role in shaping an immunosuppressive TIME. In vitro and in vivo experiments demonstrated that EMC3 promotes HCC proliferation, migration, invasion, and metastasis. Mechanistically, EMC3 activated the PI3K/AKT/mTOR signaling pathway, and inhibition of this pathway reversed the oncogenic effects of EMC3. CONCLUSIONS:EMC3 drives HCC progression by activating the PI3K/AKT/mTOR pathway and modulating the TIME. It represents a novel prognostic biomarker and a potential therapeutic target for HCC, with implications for patient stratification and personalized treatment strategies.

Background Liver fibrosis is a major contributor to liver-related morbidity and mortality among individuals with previous or current hepatitis B virus (HBV) exposure. Nutritional and inflammatory factors have been implicated in liver fibrosis, however their combined value for fibrosis stage classification remains insufficiently explored. This study aimed to investigate the associations of nutritional and inflammatory indicators with liver fibrosis severity and to develop machine-learning models for fibrosis stage classification. Methods This cross-sectional study analyzed data from the National Health and Nutrition Examination Survey (NHANES) 2017–2018 and 2019–2020 cycles. Individuals with evidence of previous or current HBV exposure (anti-HBc positive) were included. Liver fibrosis severity was assessed using transient elastography-derived liver stiffness measurements. Associations between clinical variables and fibrosis severity were evaluated using correlation analysis, univariate regression, and restricted cubic spline models. Random forest-based feature selection was performed, and six machine-learning algorithms, including Random Forest Support Vector Machine, Extreme Gradient Boosting (XGBoost), K-Nearest Neighbor, Decision Tree, and Neural Network, were developed and evaluated using repeated 10-fold cross-validation. Model performance was assessed using accuracy, multi area of under curve, F1-score, calibration analyses and . Results A total of 911 participants were included, comprising 769 individuals without fibrosis (84.41%), 39 with F1 fibrosis (4.28%), 52 with F2 fibrosis (5.71%), and 51 with F3 fibrosis (6.00%). Correlation and regression analyses demonstrated significant associations between liver fibrosis severity and multiple nutritional and inflammatory indicators. Restricted cubic spline analyses further revealed nonlinear relationships between fibrosis severity and age, BMI, GNRI, NPAR, and SIRI. Feature selection identified 10 key variables, including GNRI, BMI, albumin, total cholesterol, CRP, NPAR, creatinine, HDL cholesterol, waist-to-height ratio, and age. Among the six machine-learning algorithms evaluated, XGBoost achieved the best overall performance, with an accuracy of 0.940, a MAUC of 0.940, and a macro-F1 score of 0.804. Conclusions Nutritional and inflammatory indicators were significantly associated with liver fibrosis severity among individuals with previous or current HBV exposure. An XGBoost model constructed from routinely available clinical variables showed good performance in fibrosis stage classification and warrants further validation in independent cohorts.

[This corrects the article DOI: 10.3892/ol.2022.13608.].

Purpose Establishing a predictive model to screen for breast cancer patients at high risk of liver metastasis enables early intervention, thereby delaying the onset of liver metastasis and enhancing both the Disease-Free Survival (DMFS) and Overall Survival (OS) of affected patients. Methods A cohort of 647 patients with pathologically confirmed breast cancer liver metastases, 588 patients with lung metastases, and 189 patients with brain metastases (verified through both pathological and imaging methods) were assembled from the Affiliated Cancer Hospital of Zhengzhou University between January 2004 and December 2023, the deadline for obtaining all data is April 5, 2024. Exclusion criteria included patients with initial distant organ metastases to the lungs or brain, those with synchronous multiple organ metastases, patients with bilateral breast cancer, and those who refused surgical intervention or had concurrent malignancies at other sites. Ultimately, a group of 417 patients with primary liver metastases was identified for the study. Results High-risk factors for breast cancer liver metastasis include hormone receptor status, molecular subtyping, lymph node involvement, KI-67 expression, local recurrence, bone metastasis, and suboptimal treatment regimens. Conclusions Hormone receptor status, molecular subtyping, lymph node involvement, KI-67 expression, local recurrence, bone metastases, and inadequate treatment emerged as significant risk factors for liver metastasis as the first distant metastasis (non-bone metastasis) in breast cancer patients. In the management of early breast cancer, tailored intensive treatment based on these risk factors may confer survival benefits to patients.

Background Hepatic ischemia-reperfusion injury (HIRI) remains a clinical challenge during liver surgery and transplantation, largely due to the lack of effective pharmacological interventions. Isolinderalactone (ILL), a sesquiterpene lactone, exhibits potent antioxidant and anti-inflammatory properties. However, its therapeutic potential for HIRI and its precise molecular targets, particularly regarding the SIRT1/IRE1α-mediated endoplasmic reticulum stress (ERS) pathway, remain to be elucidated. Objective This study aimed to determine the hepatoprotective effects of ILL against HIRI and investigate its regulatory mechanism centered on the SIRT1/IRE1α signaling axis. Methods HIRI was modeled in mice via ischemia/reperfusion (I/R) and in AML12 cells via hypoxia/reoxygenation (H/R). Cellular injury, oxidative stress, endoplasmic reticulum stress (ERS), apoptosis, and inflammatory responses were assessed using biochemical, histological, molecular, and cellular imaging techniques. The physical and functional interaction between ILL and SIRT1 was characterized through molecular docking, dynamics simulations, surface plasmon resonance, and enzymatic activity assays. The role of SIRT1 was further validated using genetic knockdown and pharmacological inhibition (EX527). Results ILL treatment significantly attenuated liver tissue damage, restored intracellular redox homeostasis, and mitigated ERS-induced inflammation and apoptosis. Mechanistically, ILL not only upregulated SIRT1 expression but also directly bound to SIRT1 to enhance its deacetylase activity, thereby suppressing the downstream IRE1α/TRAF2/JNK signaling cascade. Notably, SIRT1 silencing or pharmacological blockade with EX527 significantly blunted, albeit did not completely abolish, the protective effects of ILL, indicating that the SIRT1/IRE1α axis serves as a primary, though not exclusive, mediator of ILL's hepatoprotective activity. Conclusion ILL attenuates HIRI by simultaneously elevating SIRT1 expression and activating its deacetylase function, thereby restraining IRE1α-dependent ERS and subsequent hepatic injury. These findings highlight ILL as a promising therapeutic candidate for the management of perioperative HIRI.

The prognosis for patients diagnosed with hepatocellular carcinoma with bile duct tumor thrombus (HCC-BDTT) remains dismal, and there are presently no universally accepted treatment guidelines to address this complex condition. Long-term outcomes of liver transplantation (LT) for HCC-BDTT patients are unclear, and whether LT is a proper therapeutic option for HCC-BDTT patients remains to be determined. Therefore, we design a clinical trial to evaluate whether LT can improve recurrence-free survival (RFS) and overall survival (OS) in HCC-BDTT patients. This is an open-labeled, single-arm, prospective, multicenter and real-world study aiming to assess the survival outcomes of HCC-BDTT patients in LT. Patients will be enrolled based on histological confirmation of HCC with BDTT. The study is planned to take 4 years, 2 years for enrollment and 2 years for follow-up. We anticipate that LT confers beneficial survival outcomes for HCC-BDTT patients, specifically in terms of the pivotal parameters, such as RFS and quality of life. Upon successful completion of the trial, we will extend our monitoring over a longer follow-up time to accurately estimate important indicators such as OS. We expect that this study provides substantial evidence to refine treatment guidelines through thorough data analysis, ultimately contributing to better patient outcomes and advancing our understanding of the disease.Trial Register: Trial registered at www.clinicaltrials.gov (NCT06928415)

Cholestatic liver injury is a serious pathological process with limited treatment options. The function of fibrinogen gamma chain (FGG) in the liver is not well defined. This study aims to explore the role of FGG in the progression of cholestatic liver fibrosis, and to find a new target for the treatment of cholestatic liver fibrosis. A BDL induced mouse model and LX2 cell line, as well as a co-culture system of LX2 and THP-1, were used. Gene expression, cell activation and polarization were evaluated by qRT-PCR, Western Blot, RNA-seq, and immunofluorescence. 4-Octyl itaconate (OI) was used to block the M2 polarization, and an AEAA-modified lipid nanoparticle (LNP) was developed as a platform for targeted delivery of siFGG, and its therapeutic efficacy was evaluated in vitro and in vivo. In the BDL mouse, FGG expression was significantly increased and mainly localized to activated hepatic stellate cells (HSCs). In vitro experiments confirmed that FGG directly regulated the activation state of HSCs. Overexpression of FGG in mice significantly promoted the development of liver fibrosis. Further studies revealed that FGG specifically promoted macrophage polarization toward the M2 phenotype. Co-culture experiments demonstrated that FGG was a key factor driving M2 polarization of macrophages. The siFGG delivered by AEAA-LNP effectively knocked down the expression of FGG, thereby inhibiting HSC activation, disrupting the crosstalk between HSC and M2 macrophages, and ultimately significantly alleviating BDL induced liver fibrosis in vitro and in vivo. This study identifies FGG as a key fibrosis promoter that acts through simultaneous HSC activation and M2 macrophage polarization. Targeting FGG with a novel AEAA-LNP system offers a potent dual-mechanism therapy for liver fibrosis.

Renal ischemia-reperfusion (I/R) injury is a major cause of acute kidney injury and transplant dysfunction, involving endoplasmic reticulum stress. Although activating transcription factor 6 (ATF6) regulates ER stress resolution through the unfolded protein response, its specific role in renal I/R injury remains undefined. Here, we employed murine models of renal I/R and cellular hypoxia/reoxygenation (H/R) models to systematically investigate ATF6's function. Our results show that I/R injury significantly upregulates ATF6 expression, particularly in proximal tubular epithelial cells. Functionally, ATF6 activation improved renal function and attenuated inflammation, whereas its inhibition exacerbated tubular damage. Mechanistically, we demonstrated that ATF6 transcriptionally represses four and a half LIM domain protein 2 (FHL2) through direct promoter binding. FHL2 in turn interacts with TRAF6 to activate the nuclear factor kappa-B (NF-κB) pathway. ATF6 overexpression effectively counteracted FHL2-mediated NF-κB hyperactivation, establishing a protective ATF6/FHL2/NF-κB axis. These findings identify ATF6 as a key renoprotective factor and reveal mechanistic avenues for potential therapies targeting renal I/R injury and transplant complications.

Lenvatinib serves as a crucial therapeutic agent for hepatocellular carcinoma; however, its clinical application is severely restricted by poor aqueous solubility, suboptimal bioavailability, and systemic adverse effects. To overcome these limitations, we developed a novel gold-polyphenol-based self-assembled nanoplatform (ALE) via a green, one-pot synthesis strategy integrating epigallocatechin-3-gallate (EGCG), lenvatinib, and gold ions. The resulting ALE nanoparticles exhibited uniform morphology, excellent physiological stability, and an exceptional photothermal conversion efficiency (PCE) of 51.2% under 808 nm laser irradiation. In vitro studies demonstrated that ALE-mediated photothermal therapy (PTT) significantly inhibited tumor cell proliferation by inducing apoptosis through the upregulation of heat shock proteins (HSP70/90) and caspase-3 activation. Notably, the treatment also triggered a potent pro-inflammatory response, evidenced by the increased secretion of TNF-alpha, IL-6, and TGF-beta. In vivo, ALE served as an effective contrast agent for photoacoustic (PA) imaging, enabling precise tumor delineation. Furthermore, ALE-mediated PTT effectively alleviated tumor hypoxia and achieved complete tumor suppression in an H22 liver cancer xenograft model without causing systemic toxicity or organ damage. This study presents a robust, safe, and multifunctional nanoplatform that enhances the therapeutic efficacy of lenvatinib through synergistic photothermal ablation and tumor microenvironment modulation.

Growing evidence suggests a role for the gut microbiome in progression of cholangiocarcinoma (CCA), however, its diagnostic and therapeutic potential remains incompletely characterized. Here, metagenomic sequencing was performed on fecal samples (n = 785) from individuals across East, Central, and Northwestern China. Gut microbial dysbiosis in CCA was characterized by depletion of short-chain fatty acids-producing species and enrichment of potential pathobionts (Klebsiella aerogenes, Clostridium symbiosum). Diagnostic models built using species-level markers demonstrated superior performance, compared to pathway-based models, achieving area under the curve (AUC) values of 98.63% and 99.42% in the discovery cohort, with robust cross-regional validation (AUC = 80.89% and 80.43%). The model effectively distinguished CCA from hepatocellular carcinoma (AUC = 97.86%) and liver fibrosis (AUC = 98.73%) and nonalcoholic fatty liver disease (mean AUC = 96.86%). Analysis of public datasets encompassing 6847 samples across 31 studies and 11 disease states revealed moderate disease specificity influenced by biomarker overlap across conditions. Mechanistically, depleted Bifidobacterium pseudocatenulatum suppressed CCA progression, associated with inhibition of the PI3K-AKT-mTOR pathway. Collectively, this study supports the potential of fecal metagenomic signatures as a complementary noninvasive aid for CCA detection, and provides functional evidence for a candidate protective microbe.

Aminoacyl-tRNA synthetases, as pivotal enzymes in protein biosynthesis, have been linked to the development of certain diseases when specific isoforms are dysregulated. However, the involvement of leucyl-tRNA synthetase (LARS) in hepatic pathologies remains unexplored. In this study, a marked upregulation of LARS was observed in cholestatic liver tissues. Mice with liver-specific LARS knockdown manifested substantially attenuated hepatic injury, evidenced by reductions in serum alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase levels, as well as dysregulated bile acid homoeostasis, diminished hepatic necrosis, fibrosis, and hepatocyte apoptosis. Conversely, hepatic overexpression of LARS resulted in exacerbated liver injury. To mimic cholestatic injury in vitro, AML-12 cells were exposed to taurocholic acid (TCA), where it was found that LARS deficiency alleviated TCA-induced cellular damage and apoptosis. Mechanistically, bile acids were found to induce an integrated stress response (ISR) via up-regulating the phosphorylation of general control nonderepressible (GCN) 2 levels. Notably, LARS deficiency was shown to attenuate the TCA-induced GCN2 phosphorylation upregulation, while no modulatory effect was detected on GCN2 expression in the absence of TCA. Furthermore, BDL decreased the hepatic leucine contents and tRNA cahrging ratio, which was reversed by LARS knockdown. GCN2 knockdown reversed the effect of LARS overexpression on AML-12 cell viability. Collectively, these findings delineate a regulatory role for LARS in modulating cholestatic liver injury, likely through the suppression of leucine-tRNA-GCN2 activation and ISR signaling pathway.