To explore the involvement of neutrophil dysregulation in Cronkhite-Canada syndrome (CCS) and to identify serum biomarkers for its diagnosis and disease activity assessment. We performed comprehensive serum proteomic analysis using data-independent acquisition (DIA) on samples from patients with active CCS (aCCS) and healthy controls (HCs). Candidate proteins were further evaluated by parallel reaction monitoring (PRM). An independent validation cohort including cases with aCCS, remitting CCS (rCCS), and HCs was then assessed using enzyme-linked immunosorbent assay (ELISA). In addition, previously generated transcriptomic data and immunofluorescence staining of colonic polyps were used to assess local neutrophil extracellular trap (NET)-related immune changes in intestinal tissues. Proteomic screening identified 362 differentially expressed proteins, and bioinformatic analysis consistently highlighted pathways related to neutrophil activation, acute inflammation, and NET formation. Key neutrophil-associated proteins, including myeloperoxidase (MPO), lipocalin-2 (LCN2), and matrix metalloproteinase-9 (MMP-9), were significantly upregulated. PRM validated seven upregulated candidate proteins. In the independent validation cohort, compared with HCs, serum MPO-DNA complexes, LCN2, and MMP-9 were significantly elevated in aCCS patients, whereas MPO-DNA complexes and MMP-9 were significantly reduced in rCCS cases. Tissue-level transcriptomic and immunofluorescence analyses further supported NET-related immune activation in CCS colonic polyps. Receiver operating characteristic curve analysis demonstrated favorable diagnostic performance for these biomarkers. Our findings support the involvement of neutrophil dysregulation and NET-associated pathways in CCS pathobiology. Serum MPO-DNA complexes, LCN2, and MMP-9 may serve as promising non-invasive biomarkers for diagnosis and disease activity monitoring. Further studies incorporating disease control cohorts and mechanistic validation are warranted.

Objective: To investigate the effect and underlying mechanism by which taurocholic acid (TCA) promotes colorectal cancer liver metastasis (CRLM) through regulation of neutrophil extracellular trap (NET) formation. Methods: A total of 20 CRLM patients and 20 non-metastatic colorectal cancer (non-mCRC) patients admitted to Qilu Hospital of Shandong University from 2021 to 2022 were retrospectively included. The age of the CRLM patients was (57±12) years, including 11 males and 9 females; the age of the non-mCRC patients was (60±9) years, including 15 males and 5 females. Non-targeted metabolomics was employed to identify differential serum metabolites associated with liver metastasis, and the distinguish efficacy of TCA was evaluated by receiver operating characteristic (ROC) curve analysis. Neutrophils from healthy donors were isolated by density gradient centrifugation. The differentiated HL-60 (dHL-60) model was established by inducing the acute promyelocytic leukemia cell line HL-60 to differentiate into neutrophil-like cells with all-trans retinoic acid (ATRA). The expression characteristics of carcinoembryonic antigen-related cell adhesion molecule 8 (CD66b) and integrin subunit alpha M (CD11b) were detected by flow cytometry. The cell morphology and the proportion of live cells were assessed respectively by wright-giemsa staining and trypan blue staining. In neutrophils and dHL-60 models, cells were divided into a negative control group, a phorbol 12-myristate 13-acetate (PMA, 0.5 μmol/L) positive control group and different concentrations of TCA treatment groups (0.01, 0.1, 1, 10 and 100 μmol/L). The formation of the NET reticular structure after stimulation was observed by Sytox Green staining, and the content of double-stranded deoxyribonucleic acid (ds-DNA) released after stimulation was quantitatively evaluated by PicoGreen. Western blotting was used to detect the expression levels of p44/42 mitogen-activated protein kinase (p44/42 MAPK) and its phosphorylated form (p-p44/42 MAPK), mammalian target of rapamycin (mTOR) and its phosphorylated form (p-mTOR), and peptidylarginine deiminase 4 (PAD4) in dHL-60 following stimulation. NET induced by TCA were divided into a control group, a low-dose NET group (0.1 μg/ml) and a high-dose NET group (0.3 μg/ml) according to dose, and co-cultured respectively with colorectal cancer (CRC) cell lines DLD1 and HCT116. Transwell assay was used to analyze the effect of TCA-induced NET on the migration behavior of CRC cells. The expression levels of epithelial cadherin (E-cadherin), neural cadherin (N-cadherin), and vascular endothelial growth factor A (VEGFA) were detected by Western blotting in CRC cells after co-culture. Comparisons of measurement data between groups were performed using the independent samples t-test, Mann-Whitney U test, and one-way ANOVA. Comparisons of enumeration data between groups were performed using the χ2 test or Fisher's exact test. Results: The relative abundance of 64 metabolites, including TCA [M (Q1, Q3)] [4.445 (1.669, 9.579) vs 0.956 (0.649, 1.372), P<0.01], in the serum metabolic profile of CRLM patients was higher than that of non-mCRC patients (all P<0.05), while the relative abundance of 27 metabolites was lower than that of non-mCRC patients (all P<0.05). In the ROC analysis distinguishing the 2 sample groups, TCA achieved an area under the curve (AUC) of 0.873 (95%CI: 0.741-0.984). In cell identification, both peripheral blood neutrophils and dHL-60 models displayed typical neutrophil morphology: nuclear lobulation and cytoplasm rich in fine neutral granules; the positive rates of CD66b/CD11b were>90%, and the cell viability was>90%. In vitro stimulation of peripheral blood neutrophils for 1 h or 3 h, when the TCA stimulation concentration was greater than 0.01 μmol/L, the formation of NET was more than that of the negative control group (all P<0.05). After intervention of dHL-60 for 4 h and 6 h, when the TCA stimulation concentration was greater than 0.1 μmol/L, the NET reticular structure was more than that of the negative control (all P<0.05). When the TCA stimulation concentration was greater than 10 μmol/L, the ds-DNA release was higher than that of the negative control group (P<0.001). The expression levels of p44/42 MAPK, p-p44/42 MAPK, mTOR, p-mTOR, and PAD4 in the TCA treatment group were higher than those in the negative control group (P<0.001). In co-culture experiments of NET and CRC cells, after high-dose NET treatment of DLD1 cells, the expression of E-cadherin was lower than that of the control group (P<0.001). After high and low-dose NET treatment of HCT116 cells, the expression of E-cadherin was lower than that of the control group (both P<0.001). In both CRC cell lines, after high and low-doses of NET intervention, the expression of N-cadherin and VEGFA was higher than that of the control group (all P<0.001), and their migration and invasion abilities were higher than those of the control group (all P<0.001). Conclusions: CRLM patients exhibit distinct serum metabolic profiles, among which high-abundance TCA can induce NET release and thereby promote CRC cell metastasis. This process is associated with the activation of the p44/42 MAPK/mTOR signaling pathways in neutrophils and the epithelial-mesenchymal transition in CRC cells.

Invasive pulmonary aspergillosis (IPA) poses a serious threat to immunocompromised hosts, with limited therapeutic options highlighting the need for novel strategies. Coptis chinensis Franch. (CCF), a traditional Chinese herb containing antimicrobial alkaloids like berberine, was investigated for its therapeutic efficacy and immunological effects in a murine IPA model. Immunosuppressed female KM mice infected with Aspergillus fumigatus AF293 were treated with CCF or amphotericin B (AmB). CCF significantly improved survival, reduced fungal burden, and alleviated lung pathology, without inducing hepatotoxicity or nephrotoxicity. Transcriptomic profiling revealed a time-dependent immune response. Complement-related pathways were enriched at 2 days post-infection, whereas neutrophil recruitment and NET-related pathways became more prominent by day 4. Hub gene analysis identified Syk, Rac2, Ncf1, and Cybb as key nodes associated with the NADPH oxidase complex. Western blot and inhibitor experiments further supported the involvement of this pathway in CCF-mediated protection. Additionally, 16S rDNA sequencing indicated enrichment of Clostridium species in the gut microbiota of CCF-treated mice, which was positively correlated with the expression of NADPH oxidase-related genes, suggesting a potential gut-lung association. In conclusion, these findings support the antifungal efficacy of CCF in IPA and suggest that its protective effects may involve coordinated changes in complement-related responses, NADPH oxidase-associated neutrophil activity, and gut microbiota composition.

Neutrophil extracellular traps (NETs) are increasingly recognized as key effectors of innate immune activation and drivers of inflammatory tissue injury. Beyond their established role in host defense, experimental and translational studies suggest that NETs may contribute to pain amplification through mechanisms that affect the peripheral, central, and cardiovascular systems. This review synthesizes current research on the role of NETs as pain amplifiers, with a special focus on their involvement in microvascular dysfunction, immune-mediated ischemia, neuro-immune crosstalk, and autonomic dysregulation. By influencing endothelial integrity and microcirculatory function, NET-driven processes serve as common pathophysiological factors connecting cardiovascular vulnerability and neural tissue stress. Persistent NET-associated inflammatory signaling may also contribute to central sensitization and maladaptive brain&amp;ndash;heart interactions. Within this integrated framework, pain is understood not only as a localized symptom but also as a potential clinical indicator of systemic neuro-immune and neurovascular stress along the brain-heart axis. Framing NETs as upstream modifiers of neuro-immune and neurovascular disease offers a unifying perspective on pain, cardiovascular issues, and central nervous system vulnerability. This mechanism-based view may enhance risk assessment and highlight NET-related pathways as potential targets for personalized treatment in patients with complex brain&amp;ndash;heart disease interactions.

Arrhythmogenic cardiomyopathy (ACM) is a heritable nonischemic cardiomyopathy and a leading cause of sudden cardiac death. Although inflammation is a pathological hallmark of ACM, the contribution of peptidylarginine deiminase 4 (PAD4)-dependent neutrophil extracellular trap (NET) formation and myeloperoxidase (MPO) to disease progression remains poorly defined. To define the role of PAD4-dependent NETosis and MPO signaling in ACM disease progression homozygous desmoglein-2 mutant ( Dsg2 mut/mut ) mice were utilized. We employed genetic and pharmacological approaches to determine the efficacy of targeting PAD4 and MPO on cardiac function, arrhythmogenic burden, myocardial fibrosis, inflammatory signaling, and gap junction integrity. Cardiac phenotyping included echocardiography, electrocardiography, histology, inflammatory profiling, and biochemical assays. Markers of PAD4-dependent NETosis were elevated in Dsg2 mut/mut hearts as early as 4 weeks of age, prior to cardiac dysfunction. Genetic deletion of Pad4 significantly preserved left ventricular function, reduced ectopics, attenuated myocardial fibrosis, and suppressed proinflammatory and profibrotic cytokines. MPO levels were increased in Dsg2 mut/mut hearts, and genetic ablation of Mpo preserved cardiac function, reduced arrhythmic burden, prevented myocardial fibrosis, and restored connexin-43 phosphorylation and localization. Furthermore, pharmacological MPO-inhibition improved cardiac function, reduced arrhythmias, and attenuated inflammatory signaling, though myocardial fibrosis was not fully prevented. Notably, hearts from patients with ACM demonstrated increased MPO signal in both cardiomyocytes and non-cardiomyocyte populations compared with donor controls. PAD4-dependent NETosis and MPO signaling are key drivers of inflammation, fibrosis, and arrhythmogenesis in early disease onset in ACM. Targeting neutrophil-mediated pathways represents a promising therapeutic strategy to mitigate disease progression in ACM. What Is New?: PAD4-dependent NET formation is activated early in ACM and directly contributes to myocardial inflammation, fibrosis, arrhythmias, and cardiac dysfunction. Genetic ablation of Pad4 or Mpo preserves cardiac function, reduces arrhythmogenic burden, and attenuates proinflammatory and profibrotic signaling in a Dsg2 mutant model of ACM. Pharmacological inhibition of MPO improves cardiac function and electrical stability, identifying neutrophil-derived pathways as modifiable drivers of disease.What Are the Clinical Implications?: Neutrophil-mediated inflammation represents a clinically relevant mechanism in ACM that may be targeted without global immunosuppression.MPO inhibition may offer a novel disease-modifying strategy to reduce arrhythmias and preserve cardiac function in patients with ACM.Neutrophil- and NET-associated biomarkers may improve early risk stratification and therapeutic decision-making in genetically susceptible individuals.

Antiphospholipid syndrome (APS) is an acquired systemic autoimmune thrombo-inflammatory disorder characterized by venous, arterial, and microvascular thrombosis and/or pregnancy morbidity in the setting of persistent antiphospholipid antibodies. Cardiovascular disease in APS extends well beyond classic thrombosis and includes ischemic stroke, myocardial infarction, venous thromboembolism, valvular heart disease, pulmonary embolism, accelerated atherosclerosis, and microvascular ischemic injury. Mechanistically, APS is increasingly understood as a disorder of immunothrombosis, in which antiphospholipid antibodies-particularly anti-β2-glycoprotein I antibodies-promote endothelial dysfunction, tissue factor expression, platelet activation, complement amplification, and neutrophil extracellular trap formation. In parallel, oxidized low-density lipoprotein-β2-glycoprotein I immune complexes may link autoimmunity with plaque formation and atherothrombosis. Clinical cardiovascular risk is shaped not only by antibody profile, including lupus anticoagulant positivity, double/triple positivity, and high titers, but also by traditional atherosclerotic risk factors, systemic lupus erythematosus, hematologic manifestations, and prior thrombotic phenotype. Recent advances in classification, particularly the 2023 American College of Rheumatology/European League Against Rheumatism criteria, better capture macrovascular, microvascular, and valvular domains relevant to cardiovascular practice, although classification should not substitute for diagnosis. For secondary prevention, vitamin K antagonist therapy remains the cornerstone of thrombotic APS management. Randomized trials of direct oral anticoagulants have demonstrated excess recurrent thrombosis, especially arterial events, in high-risk APS.

Neutrophil extracellular traps (NETs) extrusion in infection and diseases: A hallway for diagnosis and prognosis.

Neutrophils are usually the first cells recruited to sites of injury or infection where they mount antimicrobial responses, including the release of neutrophil extracellular traps (NETs). Toll-like receptors play a major role in the recognition of pathogen-associated molecular patterns and all but TLR3 are expressed in neutrophils. Numerous studies have reported that LPS can trigger NET formation; in nearly all cases, however, the ligand was not purified enough to target only TLR4. There also exist isolated reports on the ability of other TLRs to induce NETs. Here we comprehensively revisited the issue of TLR-elicited NET generation using ultrapure ligands. We now report that in humans, NETs are only induced following TLR2 ligation whereas engagement of other TLRs is ineffective (despite eliciting other cellular responses). A widely used (but incompletely purified) LPS preparation potently induced NET generation by binding TLR4, TLR2, and possibly other receptors, confirming previous data from other groups. By contrast, murine NETs are formed upon either TLR2 or TLR4 engagement. Mechanistically, TLR2-triggered NET formation is controlled by signaling kinases that are mobilized early (TAK1, MEK, p38 MAPK) or belatedly (Syk, PI3K, PLCγ2); acts through endogenous factors that bind the RAGE receptor; and involves PAD4 as well as endogenous reactive oxygen species, whereas elastase is dispensable. Conversely, we provide evidence that TLR4 negatively regulates NET formation in humans. Our study shows the surprisingly restricted repertoire of TLRs that can elicit NET formation in humans, and further illustrates how this emblematic neutrophil response differs between humans and rodents.

Neutrophils rapidly deploy phagocytosis, degranulation, and neutrophil extracellular trap (NET) formation to control infections, yet exaggerated NET formation contributes to tissue injury in inflammatory disease. Because NETosis is tightly linked to the cellular redox environment, we developed MorphoMapping, an imaging flow cytometry-based pipeline that resolves neutrophil morphotypes at single-cell resolution using purely morphology-derived features. Applying MorphoMapping to cytokine-primed primary human neutrophils exposed to the heme-targeted redox modulator WF10 (tetrachlorodecaoxide) revealed a compact, treatment-specific morphological state and a reproducible redistribution of cells away from NETotic, H3Cit-associated morphotypes. Instead, WF10 enriched nuclear morphologies compatible with non-NETotic termination programs. These data indicate that defined heme-dependent redox modulation suppresses NETotic neutrophil states under priming conditions and establish MorphoMapping as a scalable approach to quantify redox-sensitive neutrophil state shifts in suspension.

Breast cancer recurrence remains a clinical challenge. The period after the treatment of the primary tumor while cancer cells that evaded initial treatment lay dormant, provides a unique window of opportunity for interventions to prevent recurrence. Specific modifiable factors such as consumption of high fat diets or elevated circulating cholesterol are associated with decreased time to recurrence. Mechanistically, oxidized cholesterol and lipid species have been implicated in the regulation of the tumor microenvironment. This suggests that consumption of food prepared under oxidizing conditions such as pan-frying, may be an underappreciated risk. Using murine models of mammary cancer dormancy, we found that a diet enriched with fat from fried, cured bacon (cfBF) decreased dormancy latency times. Resulting lesions had fewer mast cells (MCs). Loss of MCs alone resulted in reemergence from dormancy. Elevated expression of a MC gene signature in breast tumors was associated with improved progression free and overall survival, highlighting the human relevance of these findings. MCs are a major source of tissue histamine, and lesions from mice fed cfBF had decreased concentrations. Importantly, antagonists of the histamine receptor 2 (H 2 R) sparked reemergence from dormancy. H 2 R antagonists are over-the-counter drugs are taken to alleviate gastroesophageal reflux disease. Chronic treatment of mice with H 2 R-antagonists sensitized platelets towards activation and crosstalk with neutrophils, and subsequent formation of neutrophil extracellular traps (NETs). The loss of platelet or NETosis activity mitigated the H 2 R-antagonist stimulated reemergence from dormancy. Therefore, we establish a novel metastatic axis which links diet to recurrence via MCs, histaminergic signaling and NETosis: Diet -- MC -- H 2 R -- ( decreased ) Platelet Activity -- ( decreased ) Neutrophil-NETosis -- ( decreased ) Reemergence from Dormancy. Our data reveal several potential intervention strategies: lifestyle, MC stabilization, histaminergic signaling, and neutrophil and platelet activity.

Silica nanoparticles (SiNPs) are extensively produced and utilized, leading to regular human exposure in both living environmental and occupational settings. Numerous studies have confirmed the detrimental effects of SiNPs on respiratory health. Neutrophil extracellular traps (NETs), web-like networks of DNA, histones, and antimicrobial proteins that neutrophils release to trap and kill pathogens, are implicated in development of some acute and chronic lung diseases. Nevertheless, the specific role of NETs in the adverse pulmonary effects of SiNPs has not been described. Male C57BL/6 mice received intratracheal instillation of 80nm SiNPs at 3.0 and 6.0mg/kg body weight (bw), once weekly for 12 weeks, to simulate actual occupational exposure scenarios. Combined with bioinformatics analysis, this exploratory study investigated SiNPs-induced lung injury and the impact on metrics related to NETs. Subsequently, DNase I was used to explore the mitigating effect and mechanisms of promoting NETs degradation on SiNPs-induced lung injury. The DNase I was administered at a dose of 300 U per mouse via intraperitoneal injection, three times a week for 12 weeks. Exposure to SiNPs induced pulmonary injury in C57BL/6 mice. Results of bioinformatic analyses and molecular bioassay techniques revealed that NETs, integrins αLβ2 and αMβ2, CD44, TGF-β and cytokines participate in SiNPs-induced lung injury. Targeted inhibition of NETs via DNase I attenuated the overexpression of integrins, CD44 and TGF-β, thereby mitigating SiNPs-induced lung injury. Exposure to SiNPs may cause pulmonary inflammation and fibrotic injury through NETs/integrins/TGF-β and NETs/cytokines/CD44 signaling pathways. Targeting NETs degradation may offer a novel therapeutic target for SiNPs-induced lung injury and provide a potential therapeutic avenue for inflammatory and fibrotic related diseases.

Liver metastasis is a leading cause of mortality in colorectal cancer (CRC), where the inflammatory tumor microenvironment, specifically neutrophil infiltration, significantly promotes metastatic colonization. This study reveals a pro-metastatic role for alpha-1 antitrypsin (A1AT) in CRC liver metastasis via a dual mechanism involving neutrophil extracellular traps (NETs) and the transmembrane protein coiled-coil domain-containing protein 25 (CCDC25). We demonstrate that A1AT, highly expressed by a liver-metastatic CRC cell line established in a mouse model, directly induces NETs' formation. Simultaneously, intracellular A1AT binds to CCDC25, preventing its lysosomal degradation and thereby increasing its surface expression. This A1AT-mediated upregulation of CCDC25 sensitizes tumor cells to surrounding NET-DNA. Upon engagement, the ILK-RAC1-CDC42 signaling cascade activates, driving extensive cytoskeletal rearrangement and enhancing the migratory and invasive capabilities of CRC cells. Collectively, our findings elucidate a mechanism wherein tumor cells exploit the A1AT-NET-CCDC25 axis to manipulate neutrophil function and boost metastatic potential. This axis represents a critical driver of CRC liver metastasis, offering novel biomarkers and promising therapeutic targets.

Diabetic periodontitis (DP) is a prevalent clinical challenge, but the efficacy of available treatments is limited. The pathogenesis of DP is driven by hyperglycemia-induced neutrophil dysregulation, which leads to excessive formation and impaired clearance of neutrophil extracellular traps (NETs), thereby amplifying inflammation and tissue destruction. Cerium oxide nanoparticles (CeNPs) represent a promising therapeutic approach because they mimic multiple enzymes to simultaneously suppress NETosis and facilitate NET degradation. However, the nonspecific effects of unmodified CeNPs within the periodontal niche limit their regulatory precision and therapeutic potential for DP. In this study, we engineered neutrophil-elastase-binding peptide (NEBP)-modified CeNPs (NEBP-PEG-CeNPs) via a DSPE-PEG2000 coating to actively target activated neutrophils. Comprehensive characterization confirmed that this surface modification preserved the multienzyme mimetic activities of CeNPs, enabling the effective regulation of both NET formation and degradation. Using neutrophil models from both human and murine sources, we demonstrated that NEBP conjugation significantly increased cellular internalization by activated neutrophils and potently reduced NETs accumulation under high-glucose conditions. Furthermore, in a DP animal model, NEBP-PEG-CeNPs exhibited superior periodontal tissue preservation and anti-inflammatory effects because of their active targeting capability and increased bioavailability for NET regulation. This study presents a feasible and targeted nanotherapeutic strategy for DP management and provides deep insights into the rational design of CeNPs for the treatment of inflammatory diseases.

Staphylococcus aureus is an opportunistic pathogen that forms robust biofilms. This poses several challenges to the host in successfully clearing the pathogen, thus leading to chronic infections. S. aureus biofilms combat neutrophils by releasing pore-forming toxins called leukocidins. This leads to the induction of neutrophil extracellular traps (NETs) and neutrophil death. NETs are primarily composed of host DNA laced with several potent antimicrobials, including histones. While NETs entrap microbes, the nuclease produced by S. aureus degrades NETs, potentially releasing NET-associated antimicrobials. It is still unknown how S. aureus can persist in a chronic infection despite the presence of several NET-associated antimicrobials. Our studies revealed that histone H3 is sequestered by S. aureus-derived extracellular DNA (eDNA) of the biofilm matrix. This resulted in the architectural modification of the biofilm due to the formation of eDNA-H3 aggregates and protection of S. aureus within the biofilm from the potential bactericidal effects of H3.

This study investigates how cholesterol metabolism promotes metastasis in triple-negative breast cancer (TNBC). By integrating public databases, we identified a positive correlation between the cholesterol metabolism transcription factor SREBP2 and the membrane receptor CCDC25 in TNBC, and their co-high expression was strongly associated with poor patient prognosis. Mechanistically, we found that SREBP2 directly binds to the promoter region of CCDC25 and activates its transcription, upregulating its expression. Functionally, the SREBP2-CCDC25 axis enhanced TNBC cells migration and invasion and promoted neutrophil extracellular traps (NETs) formation. In vivo, SREBP2 overexpression accelerated lung metastasis in TNBC, increased levels of NETs markers and elevated CCDC25 expression in metastatic lesions. Importantly, pharmacological inhibition of the SCAP-SREBP2 pathway with Fatostatin or Lycorine suppressed CCDC25 expression, reduced NETs formation, and attenuated metastasis. Collectively, these findings define a cholesterol-driven SCAP-SREBP2-CCDC25-NETs axis that promotes TNBC metastasis and highlight this pathway as a potential therapeutic target.

Acute lung injury (ALI) and chronic inflammatory lung disorders constitute significant clinical burdens with high morbidity and mortality. Neutrophils are commonly found in lung diseases and serve as pivotal effector cells governing pulmonary inflammation. MicroRNAs (miRNAs) have emerged as critical regulators of neutrophil biology and inflammatory responses. This review summarizes current understanding of miRNA-mediated regulation of neutrophil dynamics, including granulopoiesis, recruitment, and neutrophil extracellular trap (NET) formation, and delineates the contributions of the miRNA-neutrophil axis to the pathogenesis of ALI, asthma, pulmonary fibrosis, and lung cancer. This review describes the dual roles of miRNAs, acting either as promoters or suppressors in neutrophil biology and inflammatory lung diseases, and highlights mechanistic insights into miRNA-mediated pathways influencing neutrophil-driven inflammation and tissue injury. Furthermore, we explore the therapy potential of targeting the miRNA-neutrophil axis, evaluating miRNA-based therapeutics as novel strategies to modulate neutrophil-driven pathology in lung diseases. By elucidating the miRNAs-neutrophil axis as an integrative conceptual framework, this review aims to spotlight promising avenues for developing therapies for debilitating acute and chronic respiratory disorders.

Acute liver failure (ALF) represents a critical clinical challenge with limited therapeutic options beyond liver transplantation, urgently requires novel therapeutic strategies. Here, we show that a cinnamic acid derivative, CA7, confers significant protection in mouse models of ALF. The therapeutic benefits of CA7 were evidenced by the preservation of hepatic architecture, reduction of mitochondrial damage and apoptosis, and enhanced survival rates. Using an integrated approach combining single-cell RNA sequencing and functional validation, we found that neutrophils are prominently involved in the hepatic response to CA7 treatment and represent a major cellular population through which CA7 exerts its protective effects. Mechanistically, CA7 exerts its potent hepatoprotective effects by selectively suppressing reactive oxygen species production, thereby inhibiting neutrophil extracellular trap formation, a pivotal trigger of hepatic oxidative injury. This intervention attenuated neutrophil recruitment and activation, restored redox homeostasis, and suppressed activation of the NF-κB-NLRP3-IL-1β inflammatory signaling cascade. As a consequence, CA7 effectively alleviated cytokine storm-associated inflammatory responses, prevented mitochondrial dysfunction and protected hepatocyte from apoptosis. Together, these coordinated actions underlie the robust protective effects of CA7 in ALF. Our findings suggest that CA7 acts as a promising therapeutic candidate for ALF and highlight reactive oxygen species (ROS)-dependent NETosis as an important pathogenic process in life-threatening ALF.

As central effector cells in innate immunity, neutrophils are rapidly recruited to tumors following radiotherapy (RT). However, their frequent polarization toward the pro-tumor N2 phenotype and the formation of metastasis-promoting neutrophil extracellular traps (NETs) significantly limit treatment outcomes. To reprogram these RT-recruited tumor-associated neutrophils (TANs), we develop injectable hydrogel microspheres (HMPs) co-loaded with a TLR4 agonist (lipopolysaccharide) and a PAD4 inhibitor (GSK484). The resulting L/G@HMPs effectively redirect TAN polarization toward the anti-tumor N1 phenotype, inhibit NET formation, and extend neutrophil lifespan beyond 72h. In murine tumor models, the combination of RT and L/G@HMPs triggers robust innate and adaptive immune responses, marked by substantial accumulation of N1-polarized TANs and CD8+ T cells within tumors, leading to potent tumor eradication. This study presents a hydrogel-based strategy that concurrently modulates neutrophil lifespan, phenotype, and NETs, thereby transforming tumor-promoting neutrophils into anti-tumor allies for enhanced radio-immunotherapy.

Hepatocyte-derived LRG1 primes the liver for metastasis and impairs immunotherapy.

Sepsis disrupts the physiological balance between coagulation and fibrinolysis, resulting in a state in which fibrin formation exceeds fibrin removal and drives microvascular thrombosis, organ failure, and mortality. Although an early burst of endothelial tissue-type plasminogen activator (t-PA) may transiently increase plasmin generation, this phase is rapidly eclipsed by sustained upregulation of plasminogen activator inhibitor-1 (PAI-1), dysregulated activation of thrombin-activatable fibrinolysis inhibitor, depletion of endogenous anticoagulants, and progressive endotheliopathy. Beyond inhibitor excess, emerging evidence indicates that a quantitative defect in plasminogen is a central contributor to fibrinolytic insufficiency. Neutrophil extracellular traps (NETs) contain elastase, which cleaves plasminogen into inactive fragments, reducing functional plasminogen availability and impairing fibrin-bound plasmin generation. When functional plasminogen falls below rate-limiting levels, fibrin surfaces cannot efficiently support plasmin formation, resulting in persistent microvascular fibrin deposition despite elevated D-dimer concentrations. This NET-plasminogen axis links immunothrombosis to the "fibrinolytic insufficiency phenotype observed in sepsis-induced coagulopathy and overt disseminated intravascular coagulation (DIC)." Clinically, hypofibrinolysis is characterized by high D-dimers, elevated PAI-1, reduced plasmin generation, and low fibrinolytic activity on viscoelastic testing. Multimodal assessment integrating biomarker panels and viscoelastic assays, including t-PA- or urokinase-challenged protocols, may improve risk stratification. Therapeutic strategies largely targeted coagulation; however, persistent hypofibrinolysis limits their effectiveness. Translational data demonstrate that plasminogen supplementation restores functional plasminogen levels and normalizes plasmin generation in septic patients and in experimental DIC, providing proof of concept for fibrinolysis-directed therapy. Future progress requires standardized definitions, functional fibrinolytic phenotyping, and phenotype-guided clinical trials to restore the coagulo-fibrinolytic balance in sepsis.