Impact of anticoagulants and centrifugal forces on plasma neutrophil extracellular traps assay outcomes.

The oral-immune axis: neutrophil extracellular traps mediate the link between periodontitis and autoimmune diseases.

Neutrophil extracellular traps enhance platinum resistance in ovarian cancer via SHP-1 activation.

Triple-negative breast cancer (TNBC) continues to exhibit a poor response to both immunotherapy and endocrine therapy, primarily due to its complex genetic heterogeneity and tumor immune microenvironment (TIME). Neutrophil extracellular traps (NETs) are involved in neutrophil development and degranulation in treatment-resistant tumors, particularly TNBC and metastatic cases. Using a combination of single-cell RNA sequencing (scRNA-seq) datasets, bulk mRNA sequencing data from The Cancer Genome Atlas (TCGA) cohort, microarray data from the METABRIC and GEO databases, and epigenetic sequencing resources, we comprehensively investigated the functional characteristics and prognostic potential of NET-positive tumor-associated neutrophils (NET+ TAN) in TNBC. A six-gene panel derived from NET+ TAN has been identified as a reliable diagnostic biomarker for the survival of patients with TNBC and has been validated across several independent cohorts. Notably, our findings indicate that NET+ TAN facilitate T cell reprogramming in TNBC, thereby establishing a suppressive TIME and promoting tumor progression. This study provides a comprehensive perspective on the significance of NETs in patients with TNBC, with the aim of offering genetic and epigenetic insights into the mechanisms by which NETs may transition from cold to hot tumor phenotype, as well as valuable recommendations for therapeutic strategies.

SiO2 oral exposure promotes the crosstalk between NETs and PANoptosis of lung epithelial cells by driving NOX1/ROS, triggering pneumonia in mice.

Targeting leukocytes, neutrophil extracellular traps and cytokines: A conceptual review to prevent primary graft dysfunction after lung transplantation.

Fucoidan alleviates chemotherapy-induced steatohepatitis by regulating the gut-liver axis.

Multifunctional hydrogel delivery of mesenchymal stem cell secretome suppresses neutrophil extracellular trap formation and promotes diabetic wound healing via PGE2/BMAL1 pathway.

A novel risk model incorporating MMP9, TLR8, and LILRB2 drives neutrophil extracellular trap formation and promotes immune evasion in glioma.

Neutrophil extracellular traps-inhibited nanoplatform achieves comprehensive treatment of rheumatoid arthritis at different stages.

Sanguinarine inhibits deep venous thrombosis by suppressing platelets TLR4 and reducing the release of platelets-derived HMGB1.

Spatiotemporal multi-omics profiling of secondary brain injury after intracerebral hemorrhage in an optimized autologous blood-induced mouse model with human tissue validation.

Harnessing retinoic acid metabolism-related genes to identify lung adenocarcinoma subtype and establish a risk model for predicting prognosis and drug therapy response.

Neutrophil extracellular traps (NETs) contribute to chronic obstructive pulmonary disease (COPD) pathogenesis by amplifying airway inflammation. Gasdermin D (GSDMD)-mediated pyroptosis is a critical driver of COPD progression. This study provides insights into COPD pathogenesis and provides a theoretical basis for potential therapeutic targets. Mice were exposed to cigarette smoke (CS) for 16 weeks to establish a COPD model. In vitro, alveolar macrophages (AMs) (MH-S) and alveolar epithelial cells (MLE-12) were treated with cigarette smoke extract (CSE). Subsequently, NETs were isolated from phorbol-12-myristate-13-acetate (PMA)-stimulated neutrophils. Lung histopathology, inflammatory markers, and pyroptosis-related proteins were analyzed. Co-immunoprecipitation analysis was used to verify the binding of GSDMD and ubiquitin molecules in cells. Interventions included DNase1 to degrade NET and GSDMD knockdown. In CS-exposed mice, NETs increased the levels of proinflammatory cells and mediators, and lung structure was further disrupted. Pyroptosis of AMs was increased, while phagocytosis of AMs was inhibited. However, treatment with DNAse1 partially reversed the results caused by CS exposure and NET induction. Consistently, NETs aggravated inflammatory response and pyroptosis in the CSE-induced MH-S cell model. Furthermore, NETs significantly caused an increase in ROS, which promoted the activation of GSDMD deubiquitination and subsequent pyroptosis pathway in AMs. DNase1 treatment or GSDMD silencing attenuated pyroptosis, reduced inflammatory mediators, and improved lung function. NETs aggravated CS-induced lung inflammation and injury by activating GSDMD to promote pyroptosis in AMs. Targeting GSDMD or NETs represents a novel therapeutic strategy for COPD. © 2026 The Pathological Society of Great Britain and Ireland.

Neutrophil extracellular traps (NETs) are considered as a potential mechanism contributing to inflammation in pregnancy complications. To date, no "gold standard" for NET quantification has been established. The content of NETs varies depending on the detection method. The level of NETs and their components in pregnant women with preeclampsia was assessed using cytological and immunofluorescence methods, ELISA, and flow cytometry. All applied methods showed significantly elevated levels of both NETs and their individual components in women with preeclampsia.

Heart failure (HF) is a prevalent cardiovascular condition among the elderly population, with an incidence rate that continues to rise annually, highlighting the urgent need for effective therapeutic interventions. Sustained activation of Toll-like receptor 4 (TLR4) may contribute to left ventricular dysfunction and adverse cardiac remodeling through the induction of myocardial inflammation and oxidative stress - pathological processes that closely align with the hallmark features of HF. Preclinical studies in animal models have demonstrated that TLR4 deficiency improves cardiac function in aged mice; however, the precise role and underlying mechanisms of TLR4 in human HF remain poorly understood. This study aims to test the central hypothesis that TLR4 serves as a critical molecular link between chronic inflammation and the pathophysiology of HF. HF was induced in 18-month-old male C57BL/6J mice via continuous subcutaneous infusion of isoproterenol (ISO, 30 mg/kg/day) over a period of 3 weeks. Thereafter, mice received daily intraperitoneal injections of the TLR4 inhibitor TAK-242 (2 mg/kg), deoxyribonuclease I (DNase I, 5 mg/kg), or the peptidylarginine deiminase 4 (PAD4) inhibitor GSK484 (4 mg/kg) for 7 consecutive days. Cardiac function was assessed using a ultrasound imaging system. HE staining and Masson staining were employed to evaluate myocardial pathological changes and collagen deposition. ELISA was performed to measure serum levels of myeloperoxidase-DNA (MPO-DNA), neutrophil elastase-DNA (NE-DNA), cTnI, NT-proBNP, IL-1beta, IL-6 and TNF-alpha. Immunofluorescence staining was performed to detect the co-localization levels of Ly6G with myeloperoxidase (MPO) and citrullinated histone H3 (cit-H3) in myocardial tissue, in order to assess the formation level of neutrophil extracellular traps (NETs). Western blot were utilized to determine the expression level of TLR4 protein. The expression of TLR4 was significantly upregulated in the myocardial tissue of aged HF mice. Inhibition of TLR4 not only markedly improved cardiac function but also alleviated pathological damage to myocardial tissue and reduced collagen fiber deposition. Concurrently, it also decreased the serum levels of MPO-DNA, NE-DNA, NT-proBNP, cTnI, and inflammatory factors. Moreover, the colocalization levels of Ly6G with MPO or cit-H3 in myocardial tissue was also diminished. These findings were consistent with the effects observed following DNase I and GSK484 interventions. Targeting TLR4 can mitigate inflammatory responses and enhance cardiac function in HF mice by inhibiting NETs formation. Key words Heart failure " Cardiac function " Inflammation " Toll-like receptor 4 " Neutrophil extracellular traps.

Neutrophil-driven inflammation is central to the pathogenesis of chronic obstructive pulmonary disease (COPD). Emerging evidence suggests that Ca²⁺ signaling is critical in regulating neutrophil activation, recruitment and tissue residency. In this study, we investigated the function of Na⁺/Ca²⁺ exchanger 1 (NCX1), a Ca²⁺/cation membrane transporter, in neutrophils during COPD pathogenesis. Analysis of human specimens show that NCX1 is primarily upregulated in neutrophils from patients with mixed chronic bronchitis and emphysema. Cigarette smoke exposure induces NCX1 upregulation and promotes its reverse-mode transport activity, leading to elevated intracellular Ca²⁺ levels and enhanced NETs formation. Neutrophil-specific genetic deletion of Slc8a1 or pharmacological inhibition of NCX1 reverse transport effectively suppresses Ca²⁺ influx, NETs release, and neutrophil accumulation and retention, thereby ameliorating chronic bronchitis and emphysematous changes. Collectively, our findings identify NCX1 as a regulator of Ca²⁺-dependent NETs release in neutrophils. Targeting NCX1-mediated Ca²⁺ influx or NETs formation represents a potential therapeutic strategy for neutrophilic inflammation in COPD.

Hepatocellular carcinoma (HCC) is a highly aggressive malignancy with a prognosis often limited by an immunosuppressive tumor microenvironment (TME). While cancer-associated fibroblasts (CAFs) are known to influence tumor progression, the specific mechanisms by which they modulate immune cell recruitment and function remain poorly defined. This study investigates how CAF-derived CXCL1 orchestrates a pro-tumorigenic stroma through neutrophil manipulation. The study employed in vitro co-culture systems and in vivo murine models to evaluate the impact of CXCL1-overexpressing CAFs on HCC progression. The signaling mechanisms were assessed using CXCR2 and STAT3 inhibitors, while immune surveillance was monitored via CD8+ T cell activity and natural killer (NK) cell infiltration assays. CXCL1 secreted by CAFs was found to activate neutrophils via the CXCR2-STAT3 signaling axis. This activation induces the formation of neutrophil extracellular traps (NETs), which directly facilitate tumor proliferation and metastasis. Furthermore, CXCL1-driven NETs established a potent immunosuppressive TME by significantly impairing CD8+ T cell activity and reducing NK cell infiltration. Conversely, neutralizing CXCL1 or inhibiting the CXCR2-STAT3 pathway successfully suppressed NET formation, restored anti-tumor immune cell functionality, and mitigated tumor growth. The CXCL1-CXCR2-STAT3 axis is a critical driver of stromal-immune interactions in HCC. By mediating neutrophil recruitment and NET-driven immune evasion, CAFs create a permissive environment for tumor escalation. Targeting this signaling pathway represents a promising therapeutic strategy to disrupt pro-tumorigenic neutrophil mechanisms and reinvigorate the host's anti-tumor immunity.

This review summarizes current understanding of platelet-endothelial contributions to thrombosis, emphasizing molecular crosstalk [von Willebrand factor (VWF)/ADAMTS13 balance, P-selectin, platelet glycoprotein VI (GPVI), integrins, extracellular vesicles, neutrophil extracellular traps (NETs)], high-risk clinical settings, and translational advances. Highlighting GPVI-directed therapeutics, the VWF/ADAMTS13 axis in COVID-19, and opportunities and challenges for targeting the platelet-endothelial interface. Clinical and translational studies support the safety and potential efficacy of targeting platelet-endothelial interfaces. GPVI inhibitors (Glenzocimab, Revacept) have advanced through phase I/II studies with reassuring bleeding profiles and suggest benefit in ischemic stroke and lesion-directed settings. Direct interruption of platelet-VWF interactions (Caplacizumab) is established in immune thrombotic thrombocytopenic purpura (TTP), while studies show a persistent VWF/ADAMTS13 imbalance in severe COVID-19 and inflammatory states linked to microthrombosis and worse outcomes. Antiadhesion strategies (P-selectin blockade) and modulators of immunothrombosis (NET inhibitors, targeting extracellular vesicle) are also in evaluation. Targeting platelet-endothelial crosstalk has potential to reduce pathologic thrombosis while preserving hemostasis. Clinical proof of principle exists for focused approaches (anti-VWF in TTP; P-selectin blockade in vaso-occlusion; emerging GPVI inhibitors). Priorities are: defining disease contexts and timing where interface targeting is effective; validating biomarkers (VWF/ADAMTS13 ratio, soluble P-selectin, platelet activation signatures) for patient selection; and conducting adequately powered trials with rigorous bleeding endpoints.

Heart failure (HF) is a prevalent cardiovascular condition among the elderly population, with an incidence rate that continues to rise annually, highlighting the urgent need for effective therapeutic interventions. Sustained activation of Toll-like receptor 4 (TLR4) may contribute to left ventricular dysfunction and adverse cardiac remodeling through the induction of myocardial inflammation and oxidative stress – pathological processes that closely align with the hallmark features of HF. Preclinical studies in animal models have demonstrated that TLR4 deficiency improves cardiac function in aged mice; however, the precise role and underlying mechanisms of TLR4 in human HF remain poorly understood. This study aims to test the central hypothesis that TLR4 serves as a critical molecular link between chronic inflammation and the pathophysiology of HF. HF was induced in 18-month-old male C57BL/6J mice via continuous subcutaneous infusion of isoproterenol (ISO, 30 mg/kg/day) over a period of 3 weeks. Thereafter, mice received daily intraperitoneal injections of the TLR4 inhibitor TAK-242 (2 mg/kg), deoxyribonuclease I (DNase I, 5 mg/kg), or the peptidylarginine deiminase 4 (PAD4) inhibitor GSK484 (4 mg/kg) for 7 consecutive days. Cardiac function was assessed using a ultrasound imaging system. HE staining and Masson staining were employed to evaluate myocardial pathological changes and collagen deposition. ELISA was performed to measure serum levels of myeloperoxidase-DNA (MPO-DNA), neutrophil elastase-DNA (NE-DNA), cTnI, NT-proBNP, IL-1β, IL-6 and TNF-α. Immunofluorescence staining was performed to detect the co-localization levels of Ly6G with myeloperoxidase (MPO) and citrullinated histone H3 (cit-H3) in myocardial tissue, in order to assess the formation level of neutrophil extracellular traps (NETs). Western blot were utilized to determine the expression level of TLR4 protein. The expression of TLR4 was significantly upregulated in the myocardial tissue of aged HF mice. Inhibition of TLR4 not only markedly improved cardiac function but also alleviated pathological damage to myocardial tissue and reduced collagen fiber deposition. Concurrently, it also decreased the serum levels of MPO-DNA, NE-DNA, NT-proBNP, cTnI, and inflammatory factors. Moreover, the colocalization levels of Ly6G with MPO or cit-H3 in myocardial tissue was also diminished. These findings were consistent with the effects observed following DNase I and GSK484 interventions. Targeting TLR4 can mitigate inflammatory responses and enhance cardiac function in HF mice by inhibiting NETs formation.