Search Results (480)

Familial hypercholesterolemia (FH) has been associated with an increased risk of major depressive disorder (MDD), but their shared molecular signatures remain unclear. This study aimed to identify candidate genes associated with FH-MDD co-occurrence through integrative transcriptomic analysis, machine learning, summary-data-based Mendelian randomization (SMR), and clinical validation. Transcriptomic datasets from the Gene Expression Omnibus were analyzed, including GSE6054 and GSE13985 for FH and GSE98793 for MDD. Differential expression analysis and weighted gene co-expression network analysis were used to identify shared candidate genes. Candidate biomarkers were screened using least absolute shrinkage and selection operator regression and Random Forest algorithms, followed by the construction of an exploratory nomogram. SMR analysis was performed to assess genetically supported associations between candidate-gene expression and MDD risk. Selected genes were validated by PBMC RT-qPCR in an independent four-group clinical cohort comprising healthy controls and participants with FH, MDD, or co-occurring FH and MDD. Flow cytometry was subsequently used to characterize the peripheral CD4+ T-cell profile. A total of 54 shared candidate genes were identified. CD4, MRPS21, and CRTC2 were selected by machine learning and incorporated into the nomogram, which showed good discrimination in the training set and moderate performance in external validation. Immune infiltration and enrichment analyses highlighted monocyte alterations, reduced T-cell-related signals, and enrichment of T-cell receptor and TNF-mediated pathways. SMR analysis indicated that genetically predicted higher CD4 expression was inversely associated with MDD risk, whereas RT-qPCR showed higher CD4 expression in patients with FH-MDD. Flow-cytometric analysis showed that FH-MDD participants had a higher circulating CD3+CD4+ T-cell proportion, an increased effector-memory CD4+ T-cell proportion, and a higher proportion of HLA-DR+ CD4+ T cells than participants in the comparison groups. These findings suggest that CD4 may represent a promising immune-related candidate signature associated with the co-occurrence of FH and MDD, warranting further functional validation. Full article

Malaria pathogenesis involves complex immunopathological mechanisms that hinder early diagnosis and effective treatment. This study integrates multi-omics data and experimental models to identify host-derived biomarkers and elucidate their functional roles. By combining human transcriptomic datasets, weighted gene co-expression network analysis (WGCNA), and machine learning (LASSO, SVM, RF), we identified C1QB as a key hub gene. In human data, C1QB was significantly upregulated in both training and validation cohorts (AUC 0.983 and 0.970). Single-gene GSEA and immune infiltration analyses linked C1QB to apoptosis, inflammation, and altered immune cell composition, including increased activated dendritic cells and neutrophils, and decreased naïve B cells and CD8⁺ T cells. In a murine malaria model (Plasmodium berghei ANKA), C1QB expression rose as early as day one post-infection, preceding detectable parasitemia. Immunohistochemistry revealed C1QB accumulation in the liver and spleen. Single-cell RNA sequencing in the murine model confirmed monocyte-predominant expression, and scTenifoldKnk analysis suggested its role in immune regulation. Crucially, inhibiting C1q in mice via antibody intervention alleviated malaria-induced inflammation, tissue damage, and apoptosis, indicating that C1QB/C1q actively contributes to immunopathology. AI-based drug prediction and molecular docking further supported its therapeutic potential. Collectively, our findings establish C1QB as a dual biomarker and pathogenic driver in malaria, with diagnostic and therapeutic implications. Further studies are required to validate direct target engagement and clarify upstream regulatory mechanisms. Full article

Background: In recent decades, the incidence of colorectal cancer (CRC) has been rising worldwide. CRC ranks second in cancer-related mortality. The identification of reliable biomarkers for early diagnosis and prognosis prediction, along with a deeper understanding of the underlying molecular events, holds substantial promise for improving patient outcomes. The tissue inhibitor of the metalloproteinase 1 (TIMP1) gene is overexpressed in various gastrointestinal malignancies and contributes to tumor progression. However, its role in regulating the CRC tumor immune microenvironment (TIME) and its potential as a clinically actionable prognostic biomarker remain unclear. Methods: To probe how TIMP1 acts as a prognosis-related candidate biomarker in colorectal carcinoma, TCGA-derived datasets were adopted to conduct Kaplan–Meier survival assessment. We also investigated the connection between the expression abundance of TIMP1 and the infiltration of immune populations and intratumoral lymphocytes; furthermore, immune checkpoint-related genes were systematically assessed across multiple tumor types via the TISIDB and TIMER2.0 platforms, with particular emphasis on CRC. We adopted the ESTIMATE scoring system to figure out how TIMP1 gene expression correlates with the phenotypic properties of the colorectal-cancer TIME. We relied on the limma toolkit for the screening of differential transcripts from high-TIMP1 and low-TIMP1 cohorts. Enrichment assessments covering Gene Ontology terms and Kyoto Encyclopedia of Genes and Genomes entries were then carried out to predict the potential biological pathways associated with TIMP1. We constructed the protein–protein interaction map for TIMP1-interacting partners via the STRING repository. To further explore TIMP1-correlated genes, we performed Venn diagram intersection analysis combined with Spearman’s correlation test. Finally, quantitative reverse-transcription PCR was then implemented to detect TIMP1 messenger-RNA abundance inside the RKO colorectal carcinoma cell line as well as normal colonic epithelial CCD-18Co cells, which offered in vitro experimental verification for our bioinformatic outcomes. Results: According to outcome data, TIMP1 transcripts were markedly up-regulated in CRC specimens and cell lines relative to normal samples. Elevated TIMP1 expression served as a poor-prognosis indicator for overall survival (hazard ratio [HR] = 0.43, 95% confidence interval [CI] = 0.29–0.64, p < 0.001) and disease-specific survival (HR = 0.39, 95% CI = 0.22–0.68, p = 0.001) among colorectal-carcinoma patients. TIMP1-high and TIMP1-low groups exhibited notable differences in immune cell infiltration (CD8⁺ T, macrophage, mast, neutrophil, B, monocyte, dendritic, and CD4⁺ T cells). TIMP1 expression was also significantly correlated with tumor-infiltrating lymphocytes, key immune checkpoint genes (e.g., CD274 [PD-L1] and CTLA4), and immunomodulatory chemokines (e.g., CCL3 and CCL5). Twelve TIMP1-interacting DEGs were selected: COL5A1, FN1, PRG4, and a cluster of nine MMPs (MMP1/2/3/7/8/9/11/13/14), all of which showed significant positive correlations with TIMP1 (r = 0.31–0.63, all p < 0.001). Conclusions: TIMP1 expression correlates with features of the tumor immune microenvironment and extracellular matrix remodeling in CRC, suggesting that TIMP1 shows potential as a candidate biomarker. However, its potential as a therapeutic target warrants further experimental investigation. Full article

Background and Objectives: Systemic inflammatory indices derived from routine blood tests have been proposed as potential biomarkers reflecting tumor behavior. This study aimed to evaluate their association with parameters of locoregional tumor progression in cervical cancer. Materials and Methods: This prospective observational study (CERTIFIKO) included 79 patients with cervical cancer enrolled between July 2023 and December 2025. This study was registered on ClinicalTrials.gov (NCT05673252). Pre-treatment blood samples were used to calculate the neutrophil-to-lymphocyte ratio (NLR), monocyte-to-lymphocyte ratio (MLR), platelet-to-lymphocyte ratio (PLR), systemic inflammatory response (SIR), and systemic inflammatory response index (SIRI). Tumor progression parameters included stromal infiltration, parametrial involvement, lymphovascular space invasion (LVSI), fornix involvement, lymph node status, histotype, and grading. Associations were assessed using non-parametric tests and linear regression models. Results: Stromal infiltration was significantly associated with PLR (p = 0.011), MLR (p = 0.010), and NLR (p = 0.010). Parametrial involvement was associated with MLR (p = 0.004) and NLR (p = 0.012). Diffuse LVSI was associated with PLR (p = 0.002), NLR (p = 0.006), SIR (p = 0.008), and SIRI (p = 0.034). In multivariable linear regression models, diffuse LVSI remained associated with higher PLR and NLR values, whereas parametrial involvement remained associated with higher MLR, NLR, and SIRI. No significant associations were observed for fornix involvement, histotype, or lymph node status. Conclusions: Systemic inflammatory indices were associated with parameters of locoregional tumor progression in this exploratory cohort of patients with cervical cancer. These findings warrant further evaluation in larger studies before any potential clinical application. Full article

Background: Ankylosing Spondylitis (AS) and inflammatory bowel disease (IBD) exhibit substantial pathophysiological overlap, including dysregulated innate immunity, barrier dysfunction, and Th17-mediated inflammation. However, whether Na⁺ overload-related genes (NRGs) exhibit shared transcriptional alterations in IBD and AS remains unclear. This study aimed to characterize the expression patterns of NRGs across IBD and AS and to identify candidate genes associated with both diseases. Methods: Differential expression analysis was performed to identify differentially expressed NRGs (DE-NRGs) in diseased tissues relative to normal tissues. Shared DE-NRGs between IBD and AS were screened and defined as common differentially expressed NRGs (Co-DE-NRGs). We then analyzed the correlations of these Co-DE-NRGs and explored their relationships with immune cell infiltration in target tissues. Four machine learning algorithms were applied to screen key NRGs associated with both IBD and AS. Potential therapeutic agents targeting these core biomarkers were predicted using drug–gene interaction databases, and molecular docking was conducted for further validation. Results: A total of 32 shared Co-DE-NRGs were identified for IBD and AS, with nine key regulatory NRGs recognized: CALR, CD63, CYBA, DYSF, HYOU1, IL1B, JAK1, MMP9, and STAT3. Exploratory MR analysis identified disease-specific associations between genetically predicted expression of NRGs and CD, UC, and AS. Genetically predicted STAT3 expression showed positive associations with CD and UC but an inverse association with AS and therefore did not represent a consistent risk factor across the three diseases. Furthermore, transcriptome-based drug-response analysis identified four candidate agents shared between AS and at least one IBD dataset: ciclosporin, BCL-LZH-4, BRD-K79669418, and CID-5951923. Exploratory molecular docking generated STAT3 binding poses for BCL-LZH-4 and CID-5951923, with DOCK Grid Scores of −35.321896 and −28.361128, respectively. CID-5951923 was selected for representative visualization of its predicted interaction with STAT3. Single-cell RNA-sequencing analysis identified tissue- and cell-type-specific STAT3 mRNA expression patterns in the analyzed IBD colonic and AS peripheral-blood datasets, with monocytes representing a major cell population exhibiting detected STAT3 expression in the AS dataset. Conclusions: These findings identify shared NRG-related transcriptional alterations in IBD and AS, with STAT3 emerging as a candidate gene associated with both diseases. Further experimental studies are required to determine whether these alterations reflect the involvement of NECSO and to evaluate their potential diagnostic or therapeutic relevance. Full article

Hydrogel-enabled adoptive cell therapy (ACT) offers a localized and controllable strategy for improving cellular immunotherapy in solid tumors. Hydrogels can enhance cell retention and persistence, support immune cell function within the tumor microenvironment, and reduce systemic toxicity associated with broadly delivered immune stimulants. However, delivery of living immune cells imposes practical constraints on hydrogel selection, including cytocompatible encapsulation, minimal handling and injection stress, adequate transport of oxygen and soluble cues, and an appropriate balance between local retention and timely cell egress. This review summarizes natural, synthetic, and hybrid hydrogel platforms and compares physical/supramolecular assembly, covalent and enzymatic crosslinking, and photo-crosslinking. Injectable in situ-forming depots and shear-thinning/self-healing gels are highlighted for locoregional administration. Key design and reporting dimensions of hydrogels are linked to immune cell outcomes relevant to ACT. These properties include mechanics and viscoelasticity, porosity and mass transport, degradability and remodeling, bioadhesion and extracellular matrix (ECM) mimicry, and immunogenicity versus immune shielding. Finally, a cell-type-tailored framework is presented for chimeric antigen receptor T (CAR-T), T cell receptor-engineered T (TCR-T), and tumor-infiltrating lymphocyte (TIL) products, natural killer (NK) cells, and dendritic cells (DCs) or macrophage/monocyte-derived effectors. Distinct biological requirements are used to motivate corresponding material architectures and cue presentation strategies. The review also provides quantitative reporting guidance, identifies evidence gaps for γδ T cells, and discusses in vivo validation, combination ACT strategies, and translational handling constraints. Full article

Autophagy is a crucial mechanism in the host response to intracellular bacterial pathogens during which microorganisms may undergo direct degradation in autophagolysosomes. As a highly virulent intracellular pathogen, Francisella tularensis has developed survival strategies to escape from the phagosome, replicate in the cytosol of mononuclear cells, and avoid degradation within the double-membrane vacuole during the autophagy-mediated response. The aim of this study was to investigate the role of the ATG5 autophagy protein in the host immune response to Francisella tularensis subsp. holarctica, live vaccine strain (LVS), since ATG5 plays an important role in autophagosome formation during canonical autophagy. In vitro experiments were conducted on immortalized bone marrow macrophages subjected to starvation-induced autophagy. Transgenic mice deficient in ATG5 of cells of the myeloid lineage (monocytes/macrophages and granulocytes) were used to analyze the immunological responses after intradermal infection. Cytokine levels were analyzed using Luminex, RT-qPCR, and ELISA, while inflammatory cell infiltration in the lung was analyzed by immunohistochemistry. Our results demonstrate that induced autophagy decreased bacterial replication in vitro. However, ATG5 deficiency in myeloid cells in vivo significantly diminished levels of pro-inflammatory cytokine IFN-γ in the sera, spleen, liver, and lung during Francisella infection. The attenuated pro-inflammatory response also led to significantly reduced macrophage and T cell infiltration in the lung tissue. Our findings also reveal that neutralization of IL-1β in myeloid ATG5ΔMye mice increased susceptibility to tularemia by increasing bacterial burden in organs. Full article

Background: Atherosclerosis (AS) is a chronic inflammatory vascular disease lacking reliable biomarkers for early diagnosis and risk stratification. This study aimed to identify hub genes with diagnostic potential and characterize immune microenvironment remodeling in AS. Methods: GSE43292 and GSE100927 were integrated as the training cohort (n = 168), while GSE41571, GSE120521, and GSE28829 served as independent validation cohorts (n = 48). Batch effects were corrected using the ComBat algorithm. Differentially expressed genes (DEGs) were identified using limma, followed by GO/KEGG enrichment analysis. LASSO regression and Random Forest analysis were performed to identify hub genes. A logistic regression diagnostic model was constructed and evaluated using ROC analysis, 5-fold cross-validation, and external validation. Immune infiltration was assessed using ssGSEA, and correlations between hub genes and immune cells were analyzed using Spearman correlation. Results: A total of 1349 DEGs (870 upregulated and 479 downregulated) were identified. GO and KEGG analyses demonstrated significant enrichment of immune- and inflammation-related biological processes and pathways. Seven hub genes (IBSP, XAF1, SCAMP5, SAMD9L, MYBL1, PCDH12, and CDH19) were identified through the combined application of LASSO regression and Random Forest analysis. The 7-gene logistic model achieved excellent performance in the training cohort (AUC = 0.992, 95% CI: 0.981–1.000), with a mean 5-fold cross-validation AUC of 0.984 ± 0.014, and maintained robust performance in three independent validation cohorts (AUC range: 0.952–1.000). Immune infiltration analysis revealed extensive immune microenvironment remodeling, with significantly increased infiltration of 22 of the 24 immune cell types, particularly monocytes, macrophages, and myeloid cells. Spearman correlation analysis demonstrated strong associations between hub genes, particularly SAMD9L and IBSP, and immune cell infiltration. Conclusions: This study identified a robust 7-gene diagnostic signature for AS and revealed its close association with the inflammatory immune microenvironment, providing potential biomarkers for early diagnosis and risk stratification. Full article

Gastric cancer is among the leading causes of cancer-related deaths worldwide. Modern treatment approaches include nanoparticles (NPs) designed to target cancer cells, which release a therapeutic cargo facilitating the inhibition of their expansion, thereby improving anti-tumor therapies. The success of NPs, created to deliver anticancer agents and biologically active compounds, may depend on selecting the way to target cancer cells. This study focused on examining the effects of NPs made of polylactic acid (PLA) with histamine (His) end groups and loaded with 5-fluorouracil (5-FU), a known anticancer drug (PLA-His-5-FU), on human gastric cancer AGS cells in vitro. The incubation of AGS cells with PLA-His-FU NPs resulted in diminished mitochondrial membrane potential and the induction of cell apoptosis, along with cell cycle arrest and the reduction of cell proliferation. Furthermore, the NPs tested provoked the secretion of pro-inflammatory cytokines tumor necrosis factor alpha (TNF-α) and interleukin (IL)-1β by AGS cells and induced the activation of the nuclear factor kappa B (NF-κB) signaling pathway in THP-1 blue monocytes, which indicates the ability to promote the development of a milieu for the infiltration and activation of immunocompetent cells. NPs did not increase intracellular adhesion molecule (ICAM-1) deposition on AGS cells, thus potentially preventing the distribution of cancer cells. In conclusion, PLA-His-5-FU NPs show promising anticancer activity for gastric cancer AGS cells in vitro, better than PLA-OH-5-FU, and can be used in further in vivo studies to confirm this activity. Full article

Stroke remains one of the leading causes of death and disability worldwide, yet its clinical management is constrained by substantial biological heterogeneity that single-biomarker and single-omics approaches fail to resolve. The integration of multiple molecular data layers, such as genomics, epigenomics, transcriptomics, proteomics, metabolomics, and immunomics, offers a transformative framework for investigating the underlying neuroinflammatory mechanisms of different stroke subtypes and endotypes. In this review, we synthesize the current multi-omics evidence in stroke by examining how genetic variants propagate through regulatory and immune pathways and generate measurable molecular signatures and clinically relevant biomarkers. We investigate the roles of microglia, infiltrating monocyte-derived macrophages, astrocytes, neutrophils, T cells, and endothelial cells as interacting nodes in the neuroimmune network after stroke, and analyze how spatially resolved single-cell transcriptomics illuminate state-specific programs previously undetectable in bulk tissue analyses. We discuss how proteomics and metabolomics translate these cellular programs into actionable circulating biomarkers and examine emerging evidence on blood–brain barrier disruption and neurovascular unit remodeling as multi-omics-defined targets. We then explore AI and machine learning frameworks enabling the integration of heterogeneous, high-dimensional datasets for endotype classification, patient stratification, and therapeutic response prediction. Finally, we address translational barriers, including analytical standardization, multi-ancestry generalizability, and regulatory readiness, and propose a roadmap for precision stroke medicine based on systems immunology. The core conceptual point of this review is the shift from describing omics findings in stroke cases to redefining biologically meaningful neuroinflammatory endotypes and using multi-omics to enable precision cerebrovascular medicine. Full article

Background: Rheumatoid arthritis (RA) is a persistent autoimmune condition defined by widespread synovial tissue inflammation and structural joint deterioration, with an estimated global prevalence between 0.5% and 3%. The disease predominantly targets synovial joints, resulting in progressive functional impairment when therapeutic intervention is delayed or inadequate. Objective: This review aims to comprehensively examine the contributing risk factors, underlying pathophysiological processes, and recently developed immunoengineering-based therapeutic strategies applicable to the clinical management of rheumatoid arthritis. Methods: A structured review of peer-reviewed literature was undertaken through PubMed, utilizing a targeted search strategy incorporating the terms ‘rheumatoid arthritis’ and ‘immunoengineering.’ Filters were applied to restrict results to English-language publications from peer-reviewed sources. The review emphasized studies investigating genetic susceptibility, environmental determinants, immune cell behaviour, and novel therapeutic advances in RA management. Results: Multiple interdependent risk factors underpin RA development, most notably genetic variants including HLA-DRb1 alleles, alongside demographic influences such as biological sex and advancing age, as well as obesity and pathogenic microbial exposure. These factors collectively initiate a self-amplifying inflammatory process characterized by protein citrullination and the subsequent generation of anti-citrullinated protein antibodies (ACPAs) and rheumatoid factor (RF). The ensuing immune dysregulation—driven principally by monocyte and T-lymphocyte infiltration—propagates synovial inflammation and progressively destroys cartilaginous and bony structures. Conclusions: While considerable progress has been achieved in RA pharmacotherapy, existing treatments remain constrained by systemic side effects and incomplete therapeutic responses. Emerging immunoengineering strategies offer a targeted approach to modulating the molecular and immunological milieu of affected joints, providing improved therapeutic precision. Continued investigation in this area is anticipated to yield novel clinical pathways capable of substantially enhancing patient outcomes in rheumatoid arthritis care. Full article

The treatment of intracellular bacterial infections such as Chlamydia remains a significant clinical challenge due to rising antibiotic resistance and persistent, immunopathology-driven tissue damage. Macrophages are essential for host defense; they can originate from both tissue-resident precursors and circulating monocytes. During infection, macrophages at infected sites are largely derived from monocytes that migrate and differentiate there, where they phagocytose pathogens and orchestrate immune responses. The chemokine receptor CCR2 is a key regulator of this process, yet its role beyond monocyte trafficking is not fully understood. Previous studies have shown that CCR2 deficiency impairs monocyte mobilization and exacerbates disease during Chlamydia infection, shifting immune responses away from protective Th1 immunity toward pathological Th2 and Th17 polarization. Here, we investigate how CCR2 regulates macrophage function to balance protective Th1 versus pathological Th2/Th17 immunity during Chlamydia respiratory infection. Our results show that CCR2 deficiency reduces pulmonary infiltration of Ly6C^hi and Ly6C^low monocytes and shifts macrophage differentiation away from an M1-like toward an M2-like phenotype. Mechanistically, CCR2 deficiency compromises macrophage endocytosis and survival, elevates ROS production, and activates the NLRP3 inflammasome, leading to Caspase-3/GSDME-mediated pyroptosis with increased IL-1β and IL-18, while suppressing the Caspase-1/GSDMD pathway. These findings were recapitulated in vitro using C. muridarum-stimulated Ccr2-deficient bone marrow-derived macrophages (BMDMs), which also showed impaired migration, reduced M1-like polarization, diminished endocytosis, and enhanced ROS/NLRP3/pyroptosis. Furthermore, co-culture of these BMDMs with CD4⁺ T cells revealed that Th1 differentiation was inhibited, whereas Th2 and Th17 responses were promoted. Collectively, CCR2 orchestrates monocyte–macrophage function by driving M1-like polarization and inhibiting NLRP3/Caspase-3/GSDME pyroptosis to rebalance Th1/Th2/Th17 immunity, thereby enhancing bacterial clearance while mitigating immunopathological tissue damage during Chlamydia infection. Full article

Background: Postbiotics, including cell-free supernatants and their fractions, have emerged as a safe and effective alternative to live probiotics for managing intestinal inflammation. This study investigated the protective effects of low-molecular-weight fractions (<3 kDa) of the probiotic Limosilactobacillus fermentum CECT5716 (LMW-LF) in a murine model of experimental colitis. Methods: Male C57BL/6J mice were orally administered LMW-LF for 10 days prior to colitis induction with 3% dextran sodium sulfate (DSS) for 5 days. Colonic damage was assessed via the Disease Activity Index (DAI), histology, and immunofluorescence (Ocln and Ki67). Immune cell populations were analyzed by flow cytometry, while mucosal gene expression and gut microbiota composition were evaluated using RT-qPCR and 16S rRNA sequencing, respectively. Results: LMW-LF administration significantly attenuated clinical symptoms and macroscopic colonic damage. Treatment restored epithelial barrier integrity by upregulating tight junction proteins (Tjp1) and mucin genes (Muc1-3) while normalizing DSS-induced epithelial hyperproliferation. Immunologically, LMW-LF reduced pro-inflammatory monocyte infiltration; downregulated Il6, Tnfa, and Ifng; and promoted an immunoregulatory phenotype by enhancing Ampk expression and partially restoring regulatory T cell (Treg) populations. Furthermore, LMW-LF reshaped the gut microbiota by increasing alpha diversity and promoting the enrichment of beneficial taxa, specifically Akkermansia muciniphila, which correlated with improved mucus layer preservation. Conclusions: LMW-LF is an active fraction acting across the host–microbiota axis. By integrating epithelial protection, immunomodulation, and microbial reshaping, it represents a promising dietary strategy for the management of Inflammatory Bowel Diseases. Full article

Background: Sepsis, a life-threatening organ dysfunction caused by dysregulated host responses to infection, frequently involves impaired macrophage efferocytosis that leads to apoptotic cell accumulation, secondary necrosis, and persistent inflammation. Early prognostic stratification remains challenging, as current biomarkers lack sufficient specificity and sensitivity, underscoring the urgent need for novel prognosis-related indicators. Methods: We integrated bulk transcriptomic data from a discovery cohort (GSE205672) and an independent validation cohort (GSE133822) with single-cell RNA-seq profiles of early- and late-stage sepsis (GSE167363, GSE175453). WGCNA and five consensus machine-learning algorithms were combined to screen core efferocytosis-associated genes, and expression was validated via qPCR in PBMCs from sepsis patients and CLP-induced septic mice. Results: ADAP2 was identified as the core gene achieving strict consensus across all five algorithms, with early upregulation and late depletion in sepsis, predominant expression in monocytes/macrophages—particularly M1-like and IFN-responsive subsets—and a significant correlation with efferocytosis scores and immune cell infiltration. Its expression was negatively correlated with sepsis severity (SOFA score) and showed a trend toward worse survival in patients with low ADAP2 levels. Conclusions: This multi-dimensional transcriptomic study establishes ADAP2 as a candidate biomarker with potential prognostic value in sepsis, closely linked to macrophage efferocytosis. These findings may aid early risk stratification and inform macrophage-directed immunotherapies, although prospective validation and functional studies are required. Full article

Background: Recruitment and activation of monocyte-derived macrophages (MoMFs) sustain cholangitis in primary biliary cholangitis (PBC), but whether MoMFs amplify inflammation through ferroptosis remains unclear. We defined ferroptotic programs in MoMFs and evaluated the calpain/ACSL4 axis as a regulatory and therapeutic node. Methods: We analysed a public human liver single-cell RNA sequencing (scRNA-seq) dataset and examined MoMF-associated ACSL4 and 4-hydroxynonenal (4-HNE) signals in CD11b⁺CD68⁺ cells by multiplex immunofluorescence. We used a 2OA–BSA-induced PBC-like mouse model to assess liver injury, inflammation and ferroptosis-related markers and tested Liproxstatin-1 (Lip-1), rosiglitazone (ROSI) or the calpain inhibitor PD150606. Bone marrow-derived macrophages (BMDMs) from control and PBC mice were profiled and challenged with RSL3, with or without Ferrostatin-1 (Fer-1), ROSI or PD150606. Results: MoMFs were expanded in PBC livers and showed the strongest induction of ferroptosis signatures, centered on ACSL4, with enhanced inflammatory crosstalk with cholangiocytes. Human PBC tissues showed increased CD11b⁺CD68⁺ cells positive for ACSL4 or 4-HNE. In PBC-like mice, malondialdehyde (MDA) increased and glutathione (GSH) decreased, and macrophages showed greater colocalization with ferroptosis markers; Lip-1, ROSI or PD150606 improved liver biochemistry, reduced inflammation scores and limited macrophage infiltration. PBC-derived BMDMs upregulated ACSL4 and CAPN1/2 and were more sensitive to RSL3; Fer-1, ROSI or PD150606 attenuated ferroptosis-associated molecular changes. Conclusions: MoMF ferroptosis is prominently engaged in PBC, and our findings implicate a pharmacologically tractable calpain/ACSL4 axis that may contribute to macrophage ferroptotic susceptibility and inflammatory liver injury. Full article