Platelet Activation Research Articles

Antithrombotic Properties of Anthocyanin and Protein Extracts from Sweet Potatoes (Ipomoea batatas L.): Effect of Enzymatic Inactivation Treatments.

Cardiovascular diseases are a leading cause of death worldwide, with thrombosis playing a major role. Synthetic antithrombotic therapies often have adverse effects, highlighting the need for safer, natural alternatives. This study investigates the antithrombotic potential (antiplatelet and anticoagulant activities) and hemocompatibility of protein and anthocyanin extracts from sweet potato byproducts. Since polyphenol oxidase (PPO) can impact on the biological activity of these extracts, treatments to inactivate PPO were applied with ascorbic acid immersion and bleaching. Anthocyanin extract (1mg/mL) inhibited platelet aggregation induced by ADP, collagen, and PMA. Both anthocyanin and protein extract prolonged coagulation times (aPTT, thrombin time, and fibrinogen assay) and exhibited no toxicity in hemolysis and cytotoxicity analyses. Additionally, anthocyanins reduced key platelet activation markers (P-selectin, CD63, fibrinogen). Ascorbic acid pre-treatment against PPO enhanced the biological activity of anthocyanins. This study demonstrates that sweet potato anthocyanins and proteins possess significant antithrombotic properties, offering potential as sustainable, natural alternatives to synthetic thrombosis treatments. Further studies are needed to explore their bioavailability, in vivo effects, and clinical applications.

Proteomics of arterial thrombi in acute limb ischemia.

Acute limb ischemia (ALI) is characterized bya sudden decrease in limb perfusion due to arterial occlusion. Without urgent revascularization, patients are at risk of ischemic damage and amputation. This study uses novel proteomic techniques to investigate the molecular architecture of ALI thrombi, identifying key proteins that may influence coagulation dynamics and fibrinolysis resistance. Arterial thromboemboli (n = 12) collected after revascularization procedures were analyzed and compared to in vitro clots (n = 10) generated from healthy donor blood via tissue factor‒induced coagulation. Proteins were identified and quantified via liquid chromatography‒mass spectrometry (LC-MS/MS). A comprehensive literature review of the most abundant proteins allowed for categorization based on their functional roles in fibrinolysis, red blood cell (RBC) degradation, complement activation, and platelet activation. Compared to in vitro clots, ALI clots contained 141 proteins with significantly increased abundance (fold change) and 38 with decreased abundance (p < 0.05). These include 17 fibrinolysis regulators, 8 RBC-related proteins, 6 complement proteins, and 36 platelet regulators. The antifibrinolytic protein vitronectin (VTN) was strikingly enriched (1067-fold), suggesting a substantial role in fibrinolysis resistance and clot stability. Scavengers of heme/hemoglobin, thromboinflammatory complement proteins, and platelet activators were highly abundant. This proof-of-concept study introduces novel proteomic methods for arterial thrombus analysis and identifies key proteins involved in ALI pathology. Our findings reveal a delicate balance between antifibrinolytic and profibrinolytic proteins, offering potential therapeutic targets to enhance thrombolysis and improve ALI management.

Fetuin-A increases thrombosis risk in non-alcoholic fatty liver disease by binding to TLR-4 on platelets.

Fetuin-A, a liver-derived heterodimeric plasma glycoprotein, exhibits abnormal elevation in non-alcoholic fatty liver disease (NAFLD). Plasma fetuin-A levels correlate closely with both morbidity and mortality associated with cardiovascular diseases. However, the precise influence of fetuin-A on NAFLD-related platelet activation and thrombosis remains to be elucidated. Fetuin-A directly amplified agonist-induced platelet aggregation, dense granule adenosine triphosphate release, P-selectin exposure, integrin αIIbβ3 activation, spreading, and clot retraction. Mechanistically, fetuin-A bound to platelet Toll-like receptor-4 (TLR-4), activating TLR-4/MyD88, SFK/PI3 K/AKT, cGMP/PKG, and mitogen-activated protein kinase signalling pathways to enhance platelet activation. TLR-4 specific antagonist TAK-242 and TLR-4-deficient mice confirmed the TLR-4 dependence of these effects. Oral administration of firsocostat, rosuvastatin, or pioglitazone demonstrated efficacy in alleviating thrombosis formation in NAFLD mice by reducing fetuin-A levels and attenuating platelet hyperreactivity. Notably, the fetuin-A-inhibiting antibody potently suppressed platelet activation and inhibited thrombosis formation in NAFLD mice. Administration of this antibody attenuated thromboembolism and microvascular thrombosis in NAFLD mice, thereby safeguarding the lung, heart, and brain from exacerbated tissue infarcts. Finally, a positive correlation between plasma fetuin-A concentration and platelet aggregation was observed in NAFLD patients. Fetuin-A emerges as a positive regulator of platelet hyperreactivity in NAFLD. Acting via TLR-4-dependent signalling pathways, plasma fetuin-A directly amplifies platelet activation and promotes in vivo thrombosis. Firsocostat, rosuvastatin, and pioglitazone abrogate these enhancing effects by reducing fetuin-A levels. The fetuin-A-inhibiting antibody presents potential therapeutic advantages to prevent thrombotic complications in NAFLD.

Exploring the potential biomarkers between stroke and obstructive sleep apnea by WGCNA and machine learning.

Ischemic stroke (IS) and obstructive sleep apnea (OSA) are highly prevalent disorders with significant societal and individual burdens. OSA exacerbates stroke outcomes, elevates recurrent stroke risk, and impedes functional recovery. Identifying shared biomarkers and elucidating the molecular mechanisms linking IS and OSA have been critical for developing targeted therapies and improving patient prognosis. Transcriptomic data for IS and OSA were obtained from the GEO database (GSE58294, GSE135917, GSE38792, and GSE22255). After batch-effect correction, weighted gene co-expression network analysis (WGCNA) and differential expression analysis were performed to identify disease-associated genes. Functional enrichment analysis and a protein-protein interaction network construction were conducted. Advanced machine learning algorithms-Least Absolute Shrinkage and Selection Operator (LASSO) regression and random forests-were applied to screen hub genes, followed by validation of their diagnostic performance. Patients were stratified into high-and low-expression groups based on hub genes levels, and gene set enrichment analysis (GSEA) was performed to characterize pathway activity. Integration of WGCNA and differential expression analysis revealed 112 shared differentially expressed genes (DEGs) significantly associated with IS and OSA. Enrichment analysis implicated these DEGs in critical processes, including protein ubiquitination, fatty acid metabolism, cell proliferation and apoptosis, autophagy, cyclooxygenase pathway, and chromatin remodeling. Machine learning identified DUSP1 as a central hub gene, with significantly elevated expression in both IS and OSA. Diagnostic validation demonstrated robust performance for DUSP1 (AUCs: 1.000 in GSE58294, 0.885 in GSE135917, 0.718 in GSE22255), though variability was observed in GSE38792 (AUC: 0.487). GSEA highlighted distinct pathway signatures: high DUSP1 expression correlated with activation of ribosome, spliceosome, and nucleocytoplasmic transport pathway, while suppressing Ras/Rap1 signaling, platelet activation, PI3K-AKT signaling, IL-17 signaling, and immune-related pathways (e.g., Fc gamma R-mediated phagocytosis, cytokine-cytokine receptor interaction, and B cell receptor signaling pathway). Through integrative bioinformatics and machine learning, this study identifies DUSP1 as a novel hub gene linking IS and OSA. Functional annotation reveal its involvement in shared biological pathways, offering mechanistic insights into disease pathogenesis and highlighting DUSP1 as a potential therapeutic target.

LRRC8 complexes are ATP release channels regulating platelet activation and arterial thrombosis.

LRRC8 complexes are ATP release channels regulating platelet activation and arterial thrombosis.

Direct Interleukin-6 Inhibition Blunts Arterial Thrombosis by Reducing Collagen-Mediated Platelet Activation.

Recent clinical trials demonstrated a reduction in biomarkers of thrombosis and inflammation in patients with very high cardiovascular risk treated with the anti-IL (interleukin)-6 monoclonal antibody ziltivekimab. However, if and how direct IL-6 inhibition exerts antithrombotic effects remains unknown. This translational project aimed to investigate the effect of direct IL-6 inhibition on experimental arterial thrombus formation and its underlying cellular mechanisms. Three-month-old C57BL/6J male and female mice received very low dose lipopolysaccharide for 4 weeks; in addition to lipopolysaccharide, during the fourth week, mice were randomized to receive either anti-mouse IL-6 monoclonal antibody 200 μg or IgG1 isotype control. Thrombosis of the right common carotid artery was induced by endothelial-targeted laser injury. Coagulation factors and platelet reactivity were assessed in treated mice and controls. Platelets were isolated from whole blood and their reactivity to different chemical stimuli was measured by fluorescence-activated cell sorting. Additionally, whole blood samples from patients with a history of percutaneous coronary intervention were incubated ex vivo with either ziltivekimab biosimilar or IgG1 isotype control. Platelet reactivity at rest and in response to diverse chemical stimuli was quantified by fluorescence-activated cell sorting. Mice with low-grade chronic inflammation treated with anti-IL-6 monoclonal antibody displayed significantly blunted thrombus formation, without any significant difference in coagulation factors. Ex vivo stimulation with Collagen-rP (collagen-related peptide) significantly activated platelets isolated from control mice but not those obtained from mice treated with anti-IL-6 monoclonal antibody. Similarly, platelet reactivity from patients with previous percutaneous coronary intervention fell significantly after ex vivo treatment with ziltivekimab biosimilar. Direct IL-6 inhibition blunts thrombus formation by reducing collagen-induced platelet activation. These findings offer a potential mechanistic explanation for the results observed in the RESCUE trial and support the rationale of the ongoing ZEUS trial (Ziltivekimab Cardiovascular Outcome Study).

Platelet Function in Patients with Disseminated Intravascular Coagulation: Potential Markers for Improving DIC Diagnosis?

Disseminated intravascular coagulation (DIC) is a severe complication often associated with critical illness. DIC is characterized by an uncontrolled systemic activation of the hemostatic system, leading to substantial consumption of platelets and coagulation factors. The diagnosis of DIC relies on a combination of clinical findings and laboratory results, yet DIC remains challenging to confirm, especially in early stages. This systematic review investigates the reported associations between platelet function and DIC and evaluates the potential of using platelet function markers as a supplement for DIC diagnosis. PubMed and Embase were searched for relevant literature. Human studies, which included patients with DIC and assessed platelet function using dynamic platelet function assays or soluble markers, were included. In total, 24 studies met the inclusion criteria. We found that DIC patients generally exhibit increased platelet activation in vivo, indicated by elevated plasma levels of soluble markers, while ex vivo platelet aggregation was consistently reduced compared to non-DIC patients and healthy controls; however, not all studies adjusted their results for platelet count. Soluble P-selectin was the most frequently studied plasma marker and was consistently increased in DIC patients; this was most pronounced when adjusted for platelet count. However, there was considerable heterogeneity between studies regarding both study design, patient populations, platelet function assessment, and DIC diagnosis, which complicates the comparison of findings across studies. Future studies accounting for low platelet counts in dynamic function tests are necessary to assess the role of platelet aggregation in relation to DIC.

Platelet Microvesicles as a Predictor of Thromboembolic Complications in Patients With Ovarian Cancer

BACKGROUND: Platelet microvesicles are a promising marker of oncological process activity and dynamics, which allows predicting complications in patients with cancer to guide further management. AIM: This work aimed to evaluate the role of platelet microvesicles in complication development in patients with ovarian cancer. MATERIAL AND METHODS: The prognostic role of platelet microvesicles in complication development was studied in 71 patients with ovarian cancer at the Department of Oncology of A.I. Burnasyan Federal Medical Biophysical Center (FMBC, Moscow, 2020–2023). To compare the hemostasis parameters in healthy volunteers and patients with thrombosis, a sample of patients (n = 100) in the Department of Gastric Surgery of A.I. Burnasyan FMBC during the postoperative period and a control group of healthy volunteers (donors from the Department of Blood Transfusion, n = 50) were formed. We performed thromboelastography and determined the general tendency of coagulation, functional activity of platelets and fibrinogen, activity of fibrinolysis and physical and mechanical properties of the formed clots. Platelet aggregation was evaluated using a laser platelet aggregation analyzer. Cytofluorimetric analysis was performed on a standard flow cytometer using specialized software. The obtained data were processed using Statistica 10.0 statistical package. The normality of distribution of the actual data was determined using the Shapiro–Wilk test. The median and interquartile range were used to describe the groups. Analysis of variance was performed using the Kruskal–Wallis (for independent observations) and Friedman (for repeated observations) tests. RESULTS: Patients with cancer had significantly increased levels of microvesicles in peripheral blood (43.8 × 106 per mL of plasma; p = 0.0001) compared with surgical patients. High levels of microvesicles are accompanied by platelet hyperaggregation: low platelet aggregation rates corresponded to low microvesicle levels (p = 0.001), and patients with fatal thromboembolic complications and patients with ovarian cancer were predominantly in the group with high platelet microvesicle levels (35 × 106 per mL of plasma, p = 0.0001). When evaluated as a mortality predictor, the level of microvesicles in patients with ovarian cancer was found to have a sensitivity of 61.5% (54.7–82.3) and a specificity of 93.6% (83.5–98.1). CONCLUSION: Patients with ovarian cancer have high levels of circulating procoagulant microvesicles.

HMGB1-Platelet Interactions: Mechanisms and Targeted Therapy Strategies.

Platelets serve not only as crucial hemostatic components but also as pivotal regulators of inflammatory responses, capable of interacting with diverse cell types and secreting abundant extracellular factors. Accumulating evidence demonstrates that high mobility group box 1 (HMGB1), a DNA-binding protein and critical inflammatory mediator, plays multifaceted roles in disease progression, with platelets being one cellular source of circulating HMGB1. Under pathological conditions, platelets release HMGB1 into the extracellular matrix, establishing bidirectional communication between platelets and other immune cells. Moreover, HMGB1 reciprocally activates platelets through Toll-like receptors (TLRs) and receptor for advanced glycation end-products (RAGE), facilitating platelet activation and subsequent release of regulatory factors that drive inflammation-associated pathological thrombosis. In this review, we systematically characterize the HMGB1-platelet axis and elucidate its context-dependent roles in specific disease states. The mechanistic interplay between HMGB1 signaling and platelet pathophysiology is discussed, particularly its implications for disease progression. Furthermore, we critically evaluate therapeutic strategies targeting HMGB1 developed over the past decade, while proposing future directions for dual-target interventions that simultaneously modulate HMGB1 and platelet activity to combat inflammation-driven thrombotic disorders.

Platelet Metrics In Type 2 Diabetes: A Practical Tool For Assessing Vascular Complications

Background: Chronic metabolic disorder Type 2 Diabetes Mellitus (T2DM) is characterized by hyperglycemia, which is associated with a range of complications, particularly vascular complications that significantly contribute to morbidity and mortality among patients. The ability to predict these complications early is crucial for preventive strategies and patient management. This study provides an overview of the practicality of platelet parameters for predicting vascular complications in type 2 diabetes patients, highlighting their importance in diabetes management and the potential benefits of incorporating them into clinical practice as accessible and cost-effective biomarkers for early risk assessment and improved patient management in the context of a global health challenge. Method: The study included 250 T2DM patients and 125 non-diabetic individuals. Participants were selected using convenience sampling from outpatient and inpatient departments. The diabetic group was further categorized based on the presence or absence of vascular complications. Laboratory analyses were conducted to determine HbA1C levels and platelet parameters. Data were analyzed using SPSS version 21.0, applying non-parametric methods to evaluate platelet indices, HbA1c levels, and demographic variables. Result: Elevated HbA1c, platelet count, MPV (Mean Platelet Volume), and PDW (Platelet Distribution Width) values were observed in T2DM patients with vascular complications, distinguishing them from T2DM patients without such complications and healthy subjects. The patient with Diabetes Mellitus and vascular complications exhibited an MPV of 14.44 fL, PDW of 19.21%, and a platelet count of 488.8 x 10^3/µL. These values surpass the normal ranges, suggesting heightened platelet activity, size variability, and a prothrombotic state. Conclusion: The integration of MPV and PDW platelet indices could be an economical approach to T2DM vascular risk management, potentially bettering healthcare in rural settings.

Effects of Artificial Lung and Roller Pump on Platelet Morphology and Activation During Simulated Extracorporeal Circulation.

Prolonged extracorporeal circulation time is known to cause intraoperative and postoperative bleeding, but the underlying mechanism is not fully understood. In this study, we analyzed the effects of artificial lung and roller pump on platelet morphology and activation using a simulated extracorporeal circulatory circuit. Blood was drawn from healthy volunteers in the presence of unfractionated heparin as an anticoagulant and recirculated for 180 min at 0.75 L/min with the circuit under a room temperature. After the samples were collected, platelet counts, platelet surface markers, and platelet activation markers were measured. When whole blood was circulated for 180 min, platelet counts, platelet sizes, and expressions of glycoprotein Ibα (CD42b) and glycoprotein IIb (CD41) significantly decreased after recirculation. In addition, dense bodies and α-granules within platelets were also reduced after recirculation. Adhesion to collagen and platelet aggregation were significantly reduced after recirculation compared to pro-circulation. When platelets were stimulated with A23187 pro- and post-extracorporeal circulation, the expression of P-selectin was significantly lower in CD42b- platelets than in CD42b+ platelets. On the other hand, phosphatidylserine (PS) exposure in CD42b- platelets was higher than in CD42b+ platelets with or without stimulation. It was suggested that artificial lungs and roller pumps may reduce platelet activity by causing cleavage of platelet membrane glycoproteins and release of granules within platelets.

Platelet methyltransferase-like protein 4-mediated mitochondrial DNA metabolic disorder exacerbates oral mucosal immunopathology in hypoxia

Hypoxemia is a common pathological state characterized by low oxygen saturation in the blood. This condition compromises mucosal barrier integrity particularly in the gut and oral cavity. However, the mechanisms underlying this association remain unclear. This study used periodontitis as a model to investigate the role of platelet activation in oral mucosal immunopathology under hypoxic conditions. Hypoxia upregulated methyltransferase-like protein 4 (METTL4) expression in platelets, resulting in N6-methyladenine modification of mitochondrial DNA (mtDNA). This modification impaired mitochondrial transcriptional factor A-dependent cytosolic mtDNA degradation, leading to cytosolic mtDNA accumulation. Excess cytosolic mt-DNA aberrantly activated the cGAS-STING pathway in platelets. This resulted in excessive platelet activation and neutrophil extracellular trap formation that ultimately exacerbated periodontitis. Targeting platelet METTL4 and its downstream pathways offers a potential strategy for managing oral mucosa immunopathology. Further research is needed to examine its broader implications for mucosal inflammation under hypoxic conditions.

The impact of nanostructuring on the hemocompatibility of polysulfobetaine (PSB) coated hydrogel surfaces†

A series of nanostructured polysulfobetaine (PSB) hydrogel-coated surfaces were fabricated and tested for hemocompatibility in contact with human blood. PSB films were grafted onto SiO2-coated silicon wafers or Au/quartz via photochemically induced polymerization of a sulfobetaine-based monomer (SBMA, [2-(methacryloylamino)propyl]dimethyl(3-sulfopropyl)ammonium hydroxide). An anodized aluminum oxide (AAO) membrane and latex beads (LB) were used as sacrificial template structures to synthesize polysulfobetaine nanowires (PSBAAO) and hyperporous (PSBLB) networks, respectively. Two soft sacrificial templates, a liquid crystalline medium (LC) and amide-based non-ionic deep eutectic solvent (ni-DESs) providing one-dimensional ordered arrays and flickering clusters, respectively, were utilized to grow nanofibrous (PSBLC) and mesoporous (PSBDES) polysulfobetaine film. Selective dissolution of the sacrificial templates affords the transposed pattern of the template with long-range periodicity from nano to micro scale (20 to 400 nm). Electron micrograph studies revealed nanostructured materials in the form of wires (198 ± 5 nm), cavities (300 nm) and fibers (20 ± 2 nm) when AAO, LB and LC-medium were used as templates, while the polymer films prepared from ni-DESs (PSBDES), water (PSBWAT) and methanol (PSBMeOH) were devoid of any noticeable topographical features. PSB-coated surfaces (except for PSBLB) inhibited non-specific adhesion of protein and biomolecules when presented with purified human proteins, i.e., albumin, fibrinogen, hemoglobin, or human plasma, down to 20–125 ng cm−2 as shown by the QCM studies. Interestingly, the hierarchical nanostructures in polymer films (PSBAAO and PSBLC) resisted the adsorption of albumin and hemoglobin (<20 ng cm−2), even at 50 mg mL−1 concentration. The hemocompatibility of the PSB nanostructures, analyzed after contact with human whole blood for one hour on the PSBAAO and PSBLC, revealed reduced complement activation, quantified as the generation of C3bc fragments and terminal complement sC5b-9 complex formation, in comparison to acrylate glass. The nanowires of PSBAAO showed significantly lower MPO release than the PSBWAT-onto surface, whereas no difference in platelet activation was seen between the surfaces. Compactly organized nanowires and fibers increase the water of hydration layers to strengthen the antifouling and hemocompatibility features, demonstrating the bio-inert nature of the PSB nanostructures. The inherent gelation (hydrophilicity) afforded by the PSB has substantial implications in designing bio-inert surfaces for hemocompatible devices.

TNFα impairs platelet function by inhibiting autophagy and disrupting metabolism via Syntaxin-17 downregulation.

Platelets play a dual role in hemostasis and inflammation-associated thrombosis and hemorrhage. While the mechanisms linking inflammation to platelet dysfunction remain poorly understood, our previous work demonstrated that TNFα alters mitochondrial mass, platelet activation, and autophagy-related pathways in megakaryocytes. Here, we hypothesized that TNFα impairs platelet function by disrupting autophagy, a process critical for mitochondrial health and cellular metabolism. Using human and murine models of TNFα-driven diseases, including myeloproliferative neoplasms and rheumatoid arthritis, we found that TNFα downregulates STX17, a key mediator of autophagosome-lysosome fusion. This disruption inhibited autophagy, leading to the accumulation of dysfunctional mitochondria and reduced mitochondrial respiration. These metabolic alterations compromised platelet-driven clot contraction, a process linked to thrombotic and hemorrhagic complications. Our findings reveal a mechanism by which TNFα disrupts hemostasis through autophagy inhibition, highlighting TNFα as a critical regulator of platelet metabolism and function. This study provides new insights into inflammation-associated pathologies and suggests autophagy-targeting strategies as potential therapeutic avenues to restore hemostatic balance.

Analyzing Platelet Subpopulations by Multi-color Flow Cytometry.

Platelets, or thrombocytes, are small anucleated blood cells that play a crucial role in hemostasis and thrombosis. Defects in platelet functions can cause bleeding or thrombotic events in patients with cardiovascular diseases. Therefore, it is important to characterize platelets phenotypically to be able to assign platelet subpopulations to platelet function. Stimulation of human platelets with platelet agonists and activators induces morphological and physiological changes in platelets, accompanied by changes in the surface receptor population. This leads to functionally diverse platelet subpopulations. Classically defined platelet subpopulations are resting, aggregatory, procoagulant, and apoptotic platelets. To characterize the effect of agonists on platelet subtypes, we established an assay using multi-color flow cytometry consisting of 10 different antibodies and dyes (anti-CD62P, anti-CD63, anti-CD61, anti-CD41, anti-CD42b, anti-IntegrinαIIbβ3 (clone: PAC-1), anti-CXCR4, anti-ACKR3, Annexin V, Zombie NIR). Isolated human platelets were incubated with platelet agonists and stained with specific fluorophore-conjugated antibodies and dyes. Afterwards, they were measured by flow cytometry. This allows us to define agonist-specific subtypes within the classic four subpopulations. In conclusion, the combination of activation markers (anti-CD62P, anti-CD63, anti-IntegrinαIIbβ3 (clone: PAC-1)), inflammatory markers (anti-CXCR4, anti-ACKR3) and apoptotic markers (Annexin V, Zombie NIR) that compose the 10-color flow cytometry panel described in this manuscript opens up the possibility to define further platelet subtypes that could be linked to specific platelet function. This method can be applied in basic research on platelet function and physiology, as well as in defining new platelet subtypes in disease models and patient studies.

O-GalNAc Glycosylation Activates MBL-Mediated Complement and Coagulation Cascades to Drive Organotropic Metastasis.

Liver metastasis is prevalent among patients with neuroendocrine prostate cancer (NEPC) and other types of neuroendocrine (NE) cancers, featuring with an aggressive clinical course and a dismal prognosis. However, the cellular and molecular mechanisms underlying liver-specific metastatic tropism in NE cancers remain poorly understood. Intriguingly, it is found that NEPC liver metastatic foci are frequently associated with thrombi. NEPC cells express an aberrantly elevated level of glycosyltransferase Galnt9. Notably, the Galnt9-mediated O-GalNAc glycosylation on the cell membrane of NE cancer cells, particularly on the adhesion molecule Annexin A2, activates the mannose-binding lectin (MBL) complement signaling in the liver. This cascade stimulates platelet activation and thrombus formation, ultimately facilitating hepatic metastasis of NEPC. Inhibition of O-GalNAc glycosylation or knockdown of Galnt9 demonstrates efficacy in restraining the liver metastasis of NEPC, small cell lung cancer (SCLC), and colorectal neuroendocrine cancer. These findings identify Galnt9-mediated O-GalNAc glycosylation as a targetable mechanism driving liver metastasis through activation of MBL complement and coagulation cascades across a broad spectrum of NE cancers.

Platelets in preeclampsia: an observational study of indices associated with aspirin nonresponsiveness, activation and transcriptional landscape

BackgroundPlatelets play critical roles in the pathogenesis of preeclampsia, including thrombosis, endothelial dysfunction and inflammation. However, preeclampsia-associated changes in platelet gene expression and activation at the maternal–foetal interface remain unknown. Moreover, aspirin nonresponsiveness in high-risk pregnancies underscores the need for low-cost biomarkers to identify nonresponders. Nevertheless, the changes of platelet indices in women who develop preeclampsia despite aspirin prophylaxis have not yet been evaluated. In this study, we aimed to investigate the changes in platelet indices associated with aspirin nonresponsiveness, activation state and transcriptional landscape in preeclampsia.MethodsPlatelet indices were compared between aspirin-responsive and nonresponsive women. Logistic regression analysis was performed to determine the associations between platelet indices and aspirin nonresponsiveness. Opal immunofluorescence staining was performed to evaluate the expression of platelet-specific (CD42b) and activation (CD62P) markers in placental villous and decidual tissues. RNA sequencing (RNA-seq) was performed to investigate the transcriptomic profile of platelets.ResultsA decrease in platelet count (PC) during the second trimester as well as an increase in mean platelet volume (MPV) and a lower PC/MPV ratio in the third trimester were significantly associated with the subsequent development of aspirin nonresponsiveness. We observed significantly greater expression of CD62P in the placental villous and CD42b in the decidua of the preeclamptic group than in those of the nonpreeclamptic group. Colocalization analysis of CD42b and CD62P revealed that the preeclamptic placenta and decidua presented significant platelet activation. RNA-seq analysis revealed a total of 20, 618 and 1819 transcripts in the peripheral blood, placental villous and decidua of preeclamptic women, respectively. Functional analysis revealed that the PI3K-Akt and Wnt signalling pathways were significantly enriched in the placental villous and decidua of preeclamptic patients, respectively. RT‒qPCR analysis confirmed the upregulation of FKBP5, LAMA5, FZD5 and FGG mRNA expression in preeclampsia.ConclusionsOur findings suggest that PC in the second trimester and PC, MPV and PC/MPV ratio in the third trimester may be useful for assessing aspirin nonresponsiveness in women at high risk of preeclampsia. Furthermore, our findings demonstrate that preeclampsia is associated with increased platelet activation and significant enrichment of signalling pathways involved in platelet activation.

Capillaroscopy in Systemic Autoimmune Rheumatic Diseases: A Clinical Tool Linking Diagnosis and Pathogenesis

Systemic autoimmune rheumatic diseases (SARDs) including systemic lupus erythematosus (SLE), systemic sclerosis (SSc), and idiopathic inflammatory myopathies (IIMs) are multisystemic, potentially life-threatening autoimmune diseases. These diseases are associated with the highest frequency of disease-associated morbidity and mortality among rheumatic diseases, largely because their complex pathophysiology remains poorly and incompletely understood. Mortality in SARDs is associated with profound vascular dysfunction ranging from cardiovascular disease to more discreet localized vascular complications including Raynaud’s phenomenon (RP), digital ulcers, and pulmonary arterial hypertension (PAH). These vascular complications stem from damage to endothelial cells caused by immune complex deposition, platelet activation, autoantibodies that promote thrombosis (e.g., antiphospholipid antibodies), and immune dysregulation. The relationship between endothelial cell dysfunction and SARDs was first recognized by Maurice Raynaud in the 19th-century, particularly in the context of localized digital ischemia and gangrene. RP is a frequently-encountered problem in clinical practice, with a prevalence in the general population ranging from approximately 5–18%. While most cases of RP are not associated with SARDs, patients with SARDs commonly experience RP. This underscores the importance of vasculopathy related to endothelial dysfunction in the pathogenesis of SARDs. RP is often the earliest presenting feature in up to 20% of patients with SARDs. Indeed, greater than 95% of patients with SSc experience RP.11 Patients with SLE, IIMs including anti‑synthetase syndrome (ASyS), and Sjögren’s disease are also commonly affected. Hence, a closer evaluation for microvascular changes is paramount in the clinical assessment of patients with SARDs. This article will review how nailfold video capillaroscopy is emerging as a valuable point-of-care tool for diagnosis and risk stratification by providing a window into the underlying endothelial dysfunction observed in these conditions.

Platelet-leukocyte aggregates in cardiovascular disease: prognostic significance and therapeutic potential.

Initially recognised for their role in vascular haemostasis, platelets are now understood to be critical regulators of inflammation and immune responses through complex interactions with immune cells. Key to this role is the formation of platelet-leukocyte aggregates (PLAs), with platelet-monocyte aggregates (PMAs) representing the most thoroughly studied type in cardiovascular disease. PLAs form through the binding of platelet surface P-selectin to P-selectin glycoprotein ligand on leukocytes, an early interaction that may trigger broader inflammatory cascades. Recent studies link circulating PMAs with in vivo platelet activation and elevated PMA levels in patients with high thrombogenic risk, such as those with diabetes and acute myocardial infarction. Furthermore, PMA levels correlate with disease severity and long-term cardiovascular outcomes, highlighting their potential as prognostic biomarkers for adverse cardiovascular events. Therapeutic strategies, such as antiplatelet agents and P-selectin antagonists, have demonstrated efficacy in the inhibition of PLAs formation. However, the development of selective inhibitors that preserve the haemostatic and immune functions of platelets remains an unmet clinical need. Despite significant progress, the precise biological functions and underlying mechanisms of PLAs are not fully understood. This review integrates preclinical and clinical data to provide a comprehensive overview of platelet-leukocyte biology, with a focus on the prognostic role and therapeutic potential of PLAs in cardiovascular disease.

Metabolic insights into the warfarin-mango interaction: A pilot study integrating clinical observations and metabolomics

Background and purposeWarfarin is a widely prescribed oral anticoagulant for the prevention and treatment of thromboembolic events, frequently used in patients with atrial fibrillation. However, its effectiveness is often challenged by a narrow therapeutic range and significant inter-patient variability in dosage requirements and treatment responses. Drug interactions remain a critical concern, as they heighten the risk of supratherapeutic anticoagulation. Reports of interactions between warfarin and mango have documented cases of elevated international normalized ratio (INR) following mango consumption, although the underlying molecular mechanisms remain unclear.Experimental approachThis study investigates the molecular basis of the warfarin-mango interaction using proton nuclear magnetic resonance (1H-NMR)-based metabolomics. In a pre-post design study, plasma samples were collected from patients on long-term warfarin therapy (>6 months) who exhibited supratherapeutic INR levels after consuming mango. After a two-week discontinuation of mango consumption, additional plasma samples were collected once INR levels returned to the therapeutic range.Key results and conclusionThis is the first study to utilize 1H-NMR metabolomics to explore warfarin-mango interactions, integrating clinical observations with metabolic insights. Findings suggest that a reduction in glycerol 3-phosphate may impair glycolysis, disrupting platelet activation and contributing to the elevated INR levels observed in all patients. These results underscore the potential for 1H-NMR metabolomics to elucidate drug-food interactions, advancing personalized anticoagulant management and improving patient safety.