多癌种肿瘤相关静脉血栓多组学标志物筛查与功能研究

Cancer patients are at increased risk of deep vein thrombosis and pulmonary embolism. The incidence among different groups of cancer patients varies considerably depending on clinical factors, the most important being tumor entity and stage. Biomarkers have been specifically investigated for their capacity of predicting venous thromboembolism (VTE) during the course of disease. Parameters of blood count analysis (elevated leukocyte and platelet count and decreased hemoglobin) have turned out to be useful in risk prediction. Associations between elevated levels and future VTE have been found for d-dimer, prothrombin fragment 1+2, and soluble P-selectin and also for clotting factor VIII and the thrombin generation potential. The results for tissue factor–bearing microparticles are heterogeneous: an association with occurrence of VTE in pancreatic cancer might be present, whereas in other cancer entities, such as glioblastoma, colorectal, or gastric carcinoma, this could not be confirmed. Risk assessment models were developed that include clinical and laboratory markers. In the high-risk categories, patient groups with up to a >20% VTE rate within 6 months can be identified. A further improvement in risk stratification would allow better identification of patients for primary VTE prevention using indirect or novel direct anticoagulants.

Cancer patients have an increased risk of venous thromboembolism (VTE). In this review, we summarize common and cancer type–specific pathways of VTE in cancer patients. Increased levels of leukocytes, platelets, and tissue factor–positive (TF+) microvesicles (MVs) are all potential factors that alone or in combination increase cancer-associated thrombosis. Patients with lung or colorectal cancer often exhibit leukocytosis. Neutrophils could increase VTE in cancer patients by releasing neutrophil extracellular traps whereas monocytes may express TF. Thrombocytosis is often observed in gastrointestinal, lung, breast, and ovarian cancer and this could decrease the threshold required for VTE. Soluble P-selectin has been identified as a biomarker of cancer-associated thrombosis in a general cancer population and may reflect activation of the endothelium. P-selectin expression by the endothelium may enhance VTE by increasing the recruitment of leukocytes. Studies in patients with pancreatic or brain cancer suggest that elevated levels of PAI-1 may contribute to VTE. Although elevated levels of TF+ MVs have been observed in patients with different types of cancer, an association between TF+ MVs and VTE has been observed only in pancreatic cancer. Podoplanin expression is associated with VTE in patients with brain cancer and may activate platelets. Future studies should measure multiple biomarkers in each cancer type to determine whether combinations of biomarkers can be used as predictors of VTE. A better understanding of the pathways that increase VTE in cancer patients may lead to the development of new therapies to reduce the morbidity and mortality associated with thrombosis.

… There are very few studies assessing the longitudinal changes in cancer-associated VTE biomarkers in cancer patients. A recent longitudinal study over six months in patients with …

Abstract Venous thromboembolism (VTE) is a main contributor to morbidity and mortality in cancer patients. Biomarkers with the potential to predict cancer-associated VTE are continually sought. Of these, markers of thrombin generation present a likely option. The present systematic review examines the ability of three widely used biomarkers of thrombin generation: prothrombin fragment 1.2 (F1.2), thrombin-antithrombin complex (TAT), and ex vivo thrombin generation, to predict VTE in both solid and hematologic adult cancer patients. Relevant studies were identified in the PubMed and Embase databases, and the review conformed to the Preferred Reporting Items for Systematic Reviews and Meta-Analysis guidelines. Each study was evaluated using the quality assessment tool from the National Heart, Lung, and Blood Institute. The review protocol was published on PROSPERO with identifier CRD42022362339. In total, 24 papers were included in the review: 11 reporting data on F1.2, 9 on TAT, and 12 on ex vivo thrombin generation. The quality ratings of the included studies varied from good ( n  = 13), fair ( n  = 8), to poor ( n  = 3) with a high heterogenicity. However, F1.2, TAT complex, and ex vivo thrombin generation were all found to be associated with the development of VTE. This association was most pronounced for F1.2. Furthermore, the determination of F1.2 was able to improve the precision of several established risk assessment scores. In conclusion, markers of thrombin generation were found to be elevated in cancer patients with VTE, and particularly, F1.2 was found to be a promising predictor of cancer-associated VTE.

MicroRNAs (miRNAs) are small noncoding RNAs with gene regulatory functions and are commonly dysregulated in disease states. As miRNAs are relatively stable, easily measured, and accessible from plasma or other body fluids, they are promising biomarkers for the diagnosis and prediction of cancer and cardiovascular diseases. Venous thromboembolism (VTE) is the third most common cardiovascular disease worldwide with high morbidity and mortality. The suggested roles of miRNAs in regulating the pathophysiology of VTE and as VTE biomarkers are nowadays more evidenced. Patients with cancer are at increased risk of developing VTE compared to the general population. However, current risk prediction models for cancer-associated thrombosis (CAT) perform suboptimally, and novel biomarkers are therefore urgently needed to identify which patients may benefit the most from thromboprophylaxis. This review will first discuss how miRNAs mechanistically contribute to the pathophysiology of VTE. Next, the potential use of miRNAs as predictive biomarkers for VTE in subjects without cancer is reviewed, followed by an in-depth focus on CAT. Several of the identified miRNAs in CAT were found to be differentially regulated in VTE as well, giving clues on the pathophysiology of CAT. We propose that subsequent studies should be adequately sized to determine which panel of miRNAs best predicts VTE and CAT. Thereafter, validation studies using comparable patient populations are required to ultimately unveil whether miRNAs-as standalone or incorporated into existing risk models-are promising valuable VTE and CAT biomarkers.

Abstract Background and Aims Patients with cancer have increased risk of venous thromboembolism (VTE) and bleeding. Inflammatory and cardiac biomarkers may predict these complications, but their role remains unclear. This study examined associations between two inflammatory-related markers (C-reactive protein and growth differentiation factor-15) and two cardiac markers [N-terminal pro-B-type natriuretic peptide and high-sensitivity troponin T (hs-TnT)] with VTE and clinically relevant bleeding in cancer patients. Methods A post hoc analysis of the AVERT trial, which evaluated apixaban for VTE prevention in ambulatory cancer patients with a Khorana score of ≥2, was performed. Biomarkers were measured at baseline and 1 month, with C-reactive protein also at 3 months. Fine and Gray regression, accounting for competing risk of death and adjusted for age and advanced cancer, estimated subdistribution hazard ratios (SHRs) for VTE and clinically relevant bleeding. Results Of 574 patients, 514 provided baseline samples. One- and 3-month samples were available from 454 and 447, and 378 and 364, patients without prior VTE and bleeding events, respectively. Elevated baseline growth differentiation factor-15 was associated with increased VTE risk [SHR 1.36, 95% confidence interval (CI) 1.01–1.84]. N-terminal pro-B-type natriuretic peptide (SHR 1.44, 95% CI 1.08–1.92) and C-reactive protein (SHR 1.38, 95% CI 1.07–1.76) were linked to bleeding risk. Increasing high-sensitivity troponin T from baseline to 1 month was associated with higher VTE risk (SHR 1.89, 95% CI 1.14–3.16). Nomograms were developed to estimate VTE and clinically relevant bleeding risks. Conclusions Select inflammatory-related and cardiac markers were associated with VTE and bleeding risks in cancer patients, which can be determined using developed nomograms. Prospective research is needed to confirm these findings.

Abstract Background  Cancer patients have an increased risk of venous thromboembolism (VTE). Currently, the availability of highly discriminatory prediction models for VTE in cancer patients is limited. The implementation of biomarkers in prediction models might lead to refined VTE risk prediction. In this systematic review and meta-analysis, we aimed to evaluate candidate biomarkers and their association with cancer-associated VTE. Methods  We searched Medline, EMBASE, and Cochrane Central for studies that evaluated biomarkers in adult cancer patients from inception to September 2022. We included studies reporting on VTE after a cancer diagnosis with biomarker measurements performed at a defined time point. Median/mean differences (for continuous measures) and odds ratios (for dichotomous measures) with 95% confidence intervals were estimated and pooled using random-effects models. Results  We included 113 studies in the systematic review. Of these, 50 studies were included in the meta-analysis. We identified two biomarkers at cancer diagnosis (factor VIII and time to peak thrombin), three biomarkers pre-chemotherapy (D-dimer, fibrinogen, and mean platelet volume), and one biomarker preoperatively (platelet count) that had significant median or mean differences. Additionally, we found that hemoglobin <100 g/L and white blood count >11 × 10 9 /L were significantly associated with future VTE risk only when measured at cancer diagnosis. Pre-chemotherapy neutrophil-to-lymphocyte ratio ≥3 and preoperative platelet count ≥400 × 10 9 /L were also found to be associated with future VTE risk. Conclusion  In conclusion, this study identified nine candidate blood biomarkers that may help in optimizing VTE prediction in cancer patients that should be further explored in future studies.

Venous thromboembolism (VTE; deep venous thrombosis and pulmonary embolism) is associated with a poor prognosis in most malignancies and is a major cause of death among cancer patients. Universal anticoagulation for primary thromboprophylaxis in the outpatient setting is precluded by potential bleeding complications, especially without sufficient evidence that all patients would benefit from such prophylaxis. Therefore, appropriately targeting cancer patients for thromboprophylaxis is key to reducing morbidity and perhaps mortality. Predictive biomarkers could aid in identifying patients at high risk for VTE. Possible biomarkers for VTE include C-reactive protein, platelet and leukocyte counts, D-dimer and prothrombin fragment 1+2, procoagulant factor VIII, tissue factor, and soluble P-selectin. Evidence is emerging to support the use of risk assessment models in selecting appropriate candidates for primary thromboprophylaxis in the cancer setting. Further studies are needed to optimize these models and determine utility in reducing morbidity and mortality from cancer-associated thromboembolism.

Cancer-associated thrombosis (including venous thromboembolism (VTE) and arterial events) is highly consequential for patients with cancer and is associated with worsened survival. …

Abstract Venous thromboembolism (VTE) is a frequent complication in cancer patients. Platelet activation is thought to be involved in cancer-associated VTE. Here, we determined the association between evolving markers of platelet activation (soluble P-selectin [sP-selectin], soluble CD40 ligand [sCD40L], thrombospondin-1 [TSP-1] and platelet factor-4 [PF-4]) and the development of cancer-associated VTE. A nested matched case–control study was applied within a cohort of 1779 patients with different types of cancer that had been included in the Vienna Cancer and Thrombosis Study (CATS), a prospective, observational study on patients with newly diagnosed or progressive cancer after remission. Primary endpoint is symptomatic VTE during a maximum follow-up of 2 years. Cases (patients who developed VTE during follow-up) were matched in a 1:2 ratio to controls without VTE during follow-up with respect to tumor type, stage and time of observation in the study. In total, 131 VTE cases were compared to 262 controls. In logistic regression analysis, only sP-selectin was associated with risk of VTE. The odds ratios (OR) per double increase of sP-selectin, sCD40L, TSP-1 and PF-4 were 1.66 (95% confidence interval: 1.17–2.35, p = 0.005), 1.04 (0.89–1.21, p = 0.635), 1.09 (0.90–1.32, p = 0.360) and 1.03 (0.87–1.21, p = 0.737), respectively. In conclusion, sP-selectin, but not sCD40L, TSP-1 or PF-4 were associated with risk of VTE in cancer patients in this nested case–control study.

… TF MP has also been extensively studied in glioblastoma multiforme (GBM), another malignancy associated with a high rate of venous thrombosis, and different levels of expression …

BACKGROUND The mechanisms surrounding cancer-associated venous thromboembolism (VTE) are not well characterized. AVERT, a randomized placebo controlled thromboprophylaxis study in ambulatory cancer patients, provides a unique opportunity to gain insights into thrombotic mechanism(s). METHODS All available citrated platelet-free plasma samples collected at the point of randomization from individuals enrolled in the AVERT study were evaluated for the expression of D-dimer, soluble P-selectin (sP- selectin), active plasminogen activator inhibitor 1 (aPAI-1), clot lysis time (CLT) and activated factor XIa-C1 inhibitor complex (FXIa-C1). We compared the differential expression of sP-selectin, aPAI-1, CLT and FXIa-C1 among individual tumor types with normal controls. We evaluated the impact of disease type (hematologic versus solid organ malignancy) and stage (metastatic versus non-metastatic) on individual biomarker expression. RESULTS We included 449 AVERT participants in this analysis. Baseline expression of the selected thrombosis biomarkers differed significantly by individual tumor type compared with normal controls. Levels of aPAI-1, CLT, FXIa-C1 and sP-selectin were significantly elevated in individuals with lymphoma compared to individuals with non-metastatic solid organ malignancies (p<0.05). Individuals with metastatic solid organ disease had elevated levels of D-dimer and sP-selectin compared to those with non-metastatic disease (p<0.05). CONCLUSION Among a cohort of ambulatory patients at intermediate to high risk of VTE, these exploratory findings suggest that baseline activation of coagulation and fibrinolysis pathways vary significantly by tumor type and disease stage.

… -associated venous thromboembolism in a prospective observational study. Cancer 2005;104:… High platelet count associated with venous thromboembolism in cancer patients: results …

BACKGROUND Venous thromboembolism is a common complication of cancer, but the risk of developing venous thromboembolism varies greatly among individuals and depends on numerous factors, including type of cancer. We aimed to develop and externally validate a clinical prediction model for cancer-associated venous thromboembolism. METHODS We used data from the prospective Vienna Cancer and Thrombosis Study (CATS) cohort (n=1423) to select prognostic variables for inclusion in the model. We then validated the model in the prospective Multinational Cohort Study to Identify Cancer Patients at High Risk of Venous Thromboembolism (MICA) cohort (n=832). We calculated c-indices to show how the predicted incidence of objectively confirmed venous thromboembolism at 6 months compared with the cumulative 6-month incidences observed in both cohorts. FINDINGS Two variables were selected for inclusion in the final clinical prediction model: tumour-site risk category (low or intermediate vs high vs very high) and continuous D-dimer concentrations. The multivariable subdistribution hazard ratios were 1·96 (95% CI 1·41-2·72; p=0·0001) for high or very high versus low or intermediate and 1·32 (95% CI 1·12-1·56; p=0·001) per doubling of D-dimer concentration. The cross-validated c-indices of the final model were 0·66 (95% CI 0·63-0·67) in CATS and 0·68 (0·62-0·74) in MICA. The clinical prediction model was adequately calibrated in both cohorts. INTERPRETATION An externally validated clinical prediction model incorporating only one clinical factor (tumour-site category) and one biomarker (D-dimer) predicted the risk of venous thromboembolism in ambulatory patients with solid cancers. This simple model is a considerable improvement on previous models for predicting cancer-associated venous thromboembolism, and could aid physicians in selection of patients who will likely benefit from thromboprophylaxis. FUNDING Austrian Science Fund, Austrian National Bank Memorial Fund, and participating hospitals.

Abstract Background The relationship between cancer and venous thromboembolic disease (VTD) are complex because the activated coagulation factors are not only involved in thrombosis but also in malignant processes, such as angiogenesis and metastasis. Objective To compare phenotypes of extracellular vesicles (EVs), and levels of D-dimer, soluble P-selectin (sP-selectin) and antigenic tissue factor (TF) between unprovoked VTD patients, who did not develop cancer during one-year follow-up, and those with advanced stage of cancer but not associated with VTD. Methods A prospective study in which we included 138 unprovoked VTD patients and 67 advanced cancer patients, who did not develop thrombosis. Levels of EVs of different cellular origin (platelet, endothelium and leukocyte), EVs positive for tissue factor (TF) and P-selectin glycoprotein ligand 1 were quantified by flow cytometry. D-dimer, soluble P-selectin (sP-selectin) and antigenic TF were determined by ELISA. Results TF-positive EVs, D-dimer, and sP-selectin were markedly elevated in unprovoked VTD patients compared to cancer patients without association with thrombosis. Conclusions Levels of TF-positive EVs, D-dimer and sP-selectin are able to discriminate between unprovoked VTD patients not related to cancer and cancer patients not associated with VTD. These results could lead to the application of EVs as biomarkers of both diseases. Key messages: Circulating EVs, specifically TF-positive EVs, in combination with plasmatic markers of hypercoagulable states, such as D-dimer, sP-selectin and antigen TF, are able to discriminate between cancer patients without thrombosis and patients with unprovoked VTD. Research fields could be opened. Future studies will assess if these biomarkers together serve as predicting thrombotic events in cancer populations

Supplemental Digital Content is available in the text. Objective: Pancreatic cancer activates coagulation and increases risk of venous thromboembolism (VTE). We aimed at characterizing the association of hemostatic biomarkers and VTE with mortality and chemotherapy response. Approach and Results: Pancreatic cancer patients (N=145) were included in a prospective, observational cohort study (CATS [Vienna Cancer and Thrombosis Study]). Hemostatic biomarkers (D-dimer, extracellular vesicle–tissue factor activity, prothrombin fragment 1+2, fibrinogen, factor VIII, PAI-1 [plasminogen activator inhibitor 1], sP-selectin [soluble P-selectin], thrombin generation assay) were measured at inclusion. The impact of VTE on overall survival/progression-free survival (OS/PFS) was evaluated by multistate modeling. The association of biomarkers with OS was analyzed by Cox-regression and with PFS and disease control rate in patients initiating palliative chemotherapy (n=95) by Cox-regression and logistic regression. Multivariable analysis included stage, grade, sex, age, performance status, VTE (time-dependent), vascular infiltration/compression, and tumor marker levels (carbohydrate-antigen 19-9, carcinoembryonic antigen). VTE occurrence was associated with shorter OS (transition hazard ratio, 3.40 [95% CI, 2.05–5.64]) and shorter PFS (transition hazard ratio, 2.10 [1.16–3.79]). Median post-VTE OS/PFS in months was 5.5 [2.2–6.5] and 3.0 [1.5–3.9], compared with 13.4 [9.7–16.6] and 7.5 [5.9–9.8] in patients without VTE (both P<0.001). D-dimer, extracellular vesicle–tissue factor activity, PAI-1, and sP-selectin were associated with increased mortality (hazard ratio per doubling, 1.27 [1.00–1.61]; 1.63 [1.14–2.36]; 1.25 [1.06–1.47]; 1.52 [1.05–2.20]). In patients initiating palliative chemotherapy, higher D-dimer predicted shorter PFS (hazard ratio per doubling, 1.27 [1.01–1.60]) and lower disease control rate (odds ratio per doubling, 0.59 [0.36–0.98]). Conclusions: VTE diagnosis is associated with shorter OS and PFS. Higher baseline levels of D-dimer, extracellular vesicle–tissue factor activity, PAI-1, and sP-selectin were independently prognostic for increased mortality, and D-dimer predicted response to palliative chemotherapy.

Abstract Cancer-associated thrombosis affects between 1 and 20% of all patients diagnosed with cancer and is associated with significant morbidity and a poorer prognosis. Risk assessment scores exist which include the measurement of biomarkers, and which aim to identify patients at a higher risk of developing thrombotic events, but these are poor predictors and rarely used in routine clinical practice. VEGF is a potent angiogenic factor, produced by tumour cells, and released by platelets and is essential for tumour growth and progression. It also plays a role in the promotion of thrombosis through platelet activation and adhesion, and by inducing the expression of tissue factor. Therefore, the potential of VEGF to be used as a biomarker to predict cancer-associated thrombosis requires further investigation. This study reviewed the published literature to determine whether circulating VEGF levels are associated with increased risk of venous thromboembolism in patients with cancer. PubMed and OVID databases were systematically searched according to PRISMA guidelines for relevant papers using the keywords “cancer” AND “thrombosis” AND “VEGF” up to July 2023. Inclusion and exclusion criteria were applied. Seven papers (1,528 participants) were identified and included in the meta-analysis, three of which (922 participants) measured VEGF before a thrombotic event, and the remaining four (606 participants) measured VEGF at the time of the thrombosis. Our results showed that although plasma and serum VEGF tended to be higher in those who subsequently developed thrombosis than those who did not (mean difference 70.2 pg/mL for serum, and 11.44 pg/mL for plasma VEGF, 95% CI −2.39–25.73, p = 0.10), this was not found to be statistically significant. However, analysis of VEGF following blood sampling at the time of thrombosis showed a stronger statistically significant association between increased VEGF levels and presence of thrombosis (mean difference 117.02 pg/mL for serum, and 116.6 pg/mL for plasma VEGF, 95% CI 55.42–190.82, p = 0.0004). Based on current studies, whilst it is increased at the time of thrombosis, VEGF is not effective as a predictive biomarker of CAT.

Simple Summary The duration of anticoagulant treatment for venous thromboembolism in cancer patients is not well defined. The decision to suspend or re-introduce anticoagulation is based on an individual assessment between thrombotic and hemorrhagic risk. In this study, three biomarkers have shown the ability to help the clinician in decision making. Although these results should be evaluated in further studies, they open up a possibility of change in the management of cancer associated thrombosis patients. Abstract The most appropriate duration of anticoagulant treatment for cancer-associated venous thromboembolism (CAT) remains unclear. We have conducted a prospective multicenter study in CAT patients with more than 6 months of anticoagulant treatment to predict the risk of venous thromboembolism (VTE) recurrence after anticoagulation discontinuation. Blood samples were obtained when patients stopped the anticoagulation, at 21 days and at 90 days. In each sample we assessed different coagulation-related biomarkers: D-dimer (DD), high-sensitivity C-reactive protein (hs-CRP), P-selectin (PS), phospholipids, soluble tissue factor, factor VIII and the thrombin generation test. It was evaluated 325 CAT patients and 166 patients were included in the study, mean age 64 ± 17 years. VTE recurrence until 6 months after stopping anticoagulation treatment was 9.87% [95% confidence interval (CI): 6–15]. The biomarkers sub-distribution hazard ratios were 6.32 for ratio DD basal/DD 21 days > 2 (95% CI: 1.82–21.90), 6.36 for hs-CRP > 4.5 (95% CI: 1.73–23.40) and 5.58 for PS > 40 (95% CI: 1.46–21.30) after 21 days of stopping anticoagulation. This is the first study that has identified the DD ratio, hs-CRP and PS as potential biomarkers of VTE recurrence in cancer patients after the discontinuation of anticoagulation treatment. A risk-adapted strategy may allow the identification of the optimal time to withdraw the anticoagulation in each CAT patient.

Venous thromboembolism (VTE) represents a significant health concern because of its high morbidity and mortality and is moreover characterized by high rates of recurrence. It would be useful to know biomarkers that enable early identification of patients at high or low risk of primary and recurrent VTE. Various established and novel biomarkers associated with VTE have been investigated with regard to their potential for predicting primary or recurrent VTE, for facilitating the diagnosis and for optimizing the clinical management of VTE. In this review, data on selected biomarkers (D-Dimer, soluble P-selectin, coagulation factor VIII, inflammatory markers and thrombin generation) having procoagulant properties or reflecting a prothrombotic state are summarized, and their role in clinical application is discussed.

… To date, to our knowledge, there are no studies that have examined the association of biomarkers in coagulation and fibrinolytic pathways with bleeding and deep vein thrombosis (DVT…

Comprehensive protein analyses of plasma are made possible by high-throughput proteomic screens, which may help find new therapeutic targets and diagnostic biomarkers. Patients with cancer are frequently affected by venous thromboembolism (VTE). The limited predictive accuracy of current VTE risk assessment tools highlights the need for new, more targeted biomarkers. Although coagulation biomarkers for the diagnosis, prognosis, and treatment of VTE have been investigated, none of them have the necessary clinical validation or diagnostic accuracy. Proteomics holds the potential to uncover new biomarkers and thrombotic pathways that impact the risk of thrombosis. This review explores the fundamental methods used in proteomics and focuses on particular biomarkers found in VTE and cancer-associated thrombosis.

Thrombosis remains a major cause of morbidity and mortality in cancer patients. Existing risk models fail to reliably predict venous thromboembolism (VTE), underscoring the need for more accurate predictive models. In this study, we conducted a high-throughput proteomic analysis of 1,105 plasma proteins from newly diagnosed lung and gastric cancer patients prospectively monitored for VTE development. Utilizing a Bayesian probabilistic machine learning approach, we developed a predictive model incorporating 11 protein biomarkers and five clinical parameters (age, sex, history of VTE, body mass index, and hemoglobin), which significantly outperformed the Khorana prediction model. Orthogonal validation in an external placebo cohort from a phase III trial confirmed the model’s predictive power. Further investigation into the mechanistic role of CD200 receptor 1 (CD200R1), a checkpoint receptor limiting leukocyte inflammatory response that contributed strongly to the model, showed that reduced concentrations in plasma correlated with higher D-dimer and thrombosis risk. In CD200R1-deficient mice, elevated thrombin-antithrombin complexes confirmed a prothrombotic state characterized by increased interleukin-17A (IL-17A) and endothelial inflammation. Administration of anti-IL-17A antibodies to CD200R1 deficient mice normalized thrombin generation in vivo, and a meta-analysis of human clinical IL-17A inhibitory antibody studies confirmed antithrombotic activity. These findings improve the prediction of thrombosis in cancer and highlight the utility of plasma proteomics to identify unanticipated mechanistic insights and therapeutic targets in thrombo-inflammatory disease.

The plasma levels of pro- and anticoagulant proteins are important markers for venous thrombosis (VT) risk and can be affected by both genetic and acquired factors, including cancer. Generally, these markers are measured using activity- or antibody-based assays. Targeted proteomics with stable-isotope-labeled internal standards has proven adept at the rapid, multiplex, and precise quantification of proteins in complex biological samples such as plasma. We used liquid chromatography coupled to multiple reaction monitoring (MRM) mass spectrometry to evaluate the concentrations of 31 coagulation- and fibrinolysis-related proteins in plasma from 25 healthy controls, 25 patients with VT, and 25 patients with VT who were also diagnosed with cancer. The concentration level of 1 to 3 proteotypic peptides per protein was determined, and all samples were previously characterized using traditional antibody- or activity-based methods. When comparing the conventional and the MRM strategies, the mean Pearson correlation for the 13 proteins (covered by 36 target peptides) shared between the 2 approaches was 0.77, indicating a good correlation. Additionally, MRM offers higher sensitivity (mean regression slope, 0.81), higher multiplicity in a single run, and good ability to leverage all measurements to discriminate groups using unsupervised clustering, which identified vitamin K antagonist users as well as patients with VT and cancer. The data collected using MRM show that the combination of coagulation factor levels yields signature information on VT and cancer, which was not obvious from a single measurement. These results encourage the further validation and investigation of MRM in profiling protein signature of disease.

Simple Summary The risk of venous thromboembolism in cancer is nine times higher than in the general population and the second leading cause of death in these patients. Tissue factor and downstream plasmatic coagulation cascade are largely responsible for the risk of thrombosis in cancer. In recent years, it has been increasingly recognised that platelets also play a central role in tumour growth and cancer-associated thrombosis. The underlying molecular mechanisms are largely unknown. In order to comprehensively investigate the biochemical changes in platelets from cancers with high risk of thrombosis, we examined the platelet proteome of brain and lung cancer patients in comparison to sex and age-matched healthy controls. However, we only found alterations in lung cancer, where some of these platelet proteins directly promote thrombosis. One example is the increased amount of the enzyme protein disulfide isomerase, which is clinically investigated as an antithrombotic drug target of the plant-based flavonol quercetin. Abstract In order to comprehensively expose cancer-related biochemical changes, we compared the platelet proteome of two types of cancer with a high risk of thrombosis (22 patients with brain cancer, 19 with lung cancer) to 41 matched healthy controls using unbiased two-dimensional differential in-gel electrophoresis. The examined platelet proteome was unchanged in patients with brain cancer, but considerably affected in lung cancer with 15 significantly altered proteins. Amongst these, the endoplasmic reticulum (ER) proteins calreticulin (CALR), endoplasmic reticulum chaperone BiP (HSPA5) and protein disulfide-isomerase (P4HB) were significantly elevated. Accelerated conversion of the fibrin stabilising factor XIII was detected in platelets of patients with lung cancer by elevated levels of a coagulation factor XIII (F13A1) 55 kDa fragment. A significant correlation of this F13A1 cleavage product with plasma levels of the plasmin–α-2-antiplasmin complex and D-dimer suggests its enhanced degradation by the fibrinolytic system. Protein association network analysis showed that lung cancer-related proteins were involved in platelet degranulation and upregulated ER protein processing. As a possible outcome, plasma FVIII, an immediate end product for ER-mediated glycosylation, correlated significantly with the ER-executing chaperones CALR and HSPA5. These new data on the differential behaviour of platelets in various cancers revealed F13A1 and ER chaperones as potential novel diagnostic and therapeutic targets in lung cancer patients.

Background: Existing risk models for cancer-associated thrombosis (CAT) show suboptimal performance in selective high-risk populations with cancer. Affinity-based plasma proteomics offer a novel approach for detecting CAT risk. Objectives: To identify plasma biomarkers for CAT using proximity extension assays in an advanced cancer cohort. Patients/Methods: We performed a nested case-control study using the Olink Explore HT panel. The final cohort included 57 patients with CAT and 113 matched control patients from five selected cancer types who had samples collected between cancer diagnosis and chemotherapy initiation. Random survival forest model was used to assess non-linear associations with CAT in 5,416 normalized protein expressions and 8 clinical variables. Evaluation metrics averaged across bootstrapped out-of-bag test sets included time-dependent receiver operation characteristic curve (TD-ROC), calibration plot, and cumulative incidence in high- versus low-risk predicted groups. We used SHapley Additive exPlanation (SHAP) for feature interpretability. We performed overrepresentation analysis (ORA) and gene set enrichment analysis (GSEA) to assess biological pathway plausibility. Results: Our internally validated model predicted early thrombotic events well (TD-ROC 0.83 at 30 days and 0.73 at 90 days), but the discrimination waned with follow-up time (0.67 at 180 days). Calibration followed a similar pattern. In ORA and GSEA, important proteins were observed in hemostatic pathways including platelet activation, fibrin clot formation, and complement cascade regulation. Conclusion: Affinity-based plasma proteomics can be used as a novel strategy to identify biomarkers of CAT. External validation with larger sample size in a cohort setting is required for risk prediction models.

Cancer patients have an eleven-fold increased risk of venous thromboembolism (VTE) compared to the general population. In this hypothesis-generating study we investigated plasma protein levels in colorectal cancer patients with and without thrombosis using targeted absolute quantification of 269 plasma proteins. We included samples from 142 patients with stage III/IV colorectal cancer from MICA - a multinational prospective cohort study, and from 98 patients from CATS - a prospective cohort study with stage III/IV colorectal cancer. The primary outcome was objectively confirmed symptomatic or incidental deep vein thrombosis or pulmonary embolism during a 6-month follow-up period. In MICA, 11 (7.7%) developed VTE; in CATS 6 patients (6.1%) developed VTE within 6 months, and 10 (10.2%) within 2 years. Six differentially abundant proteins (DAPs) were identified in MICA: APOB100, CD5L, IGHG1, IGHM, PRG4, and TF. A model using these six proteins achieved a c-statistic of 0.687 (95% CI: 0.658-0.717) by internal cross-validation with 100 repeats and random 80% training sets. The optimism-corrected c-statistics was 0.675 (95% CI: 0.648-0.701). Using any subset of the DAPs yielded a c-statistics >0.67, with the best model reaching 0.768 (95% CI: 0.746-0.790). This outperformed the Khorana, modified-Vienna, PROTECHT, and CONKO models. The six DAPs had also predictive value in CATS with c-statistics at 0.70 (95% CI:0.671-0.728) for the 6-month, and 0.73 (95% CI:0.702-0.728) for 2-year follow-up. Two proteins showed inverted patterns between cohorts, likely due to chemotherapy. Our findings call for further investigation into the proteins identified, warranting further validation in larger, standardized studies.

Objective Non-small cell lung cancer (NSCLC) patients present a high incidence of venous thromboembolism (VTE) with poor prognosis. It is crucial to identify and diagnose VTE early. The study aimed to identify potential protein biomarkers and mechanism of VTE in NSCLC patients via proteomics research. Methods Proteomic analysis of the human plasma was performed through data-independent acquisition mass spectrometry for 20 NSCLC patients with VTE, and 15 NSCLC patients without VTE. Significantly differentially expressed proteins were analyzed by multiple bioinformatics method for further biomarker analysis. Results A total of 280 differentially expressed proteins were identified in VTE and non-VTE patients, where 42 were upregulated and 238 were downregulated. These proteins were involved in acute-phase response, cytokine production, neutrophil migration and other biological processes related to VTE and inflammation. Five proteins including SAA1, S100A8, LBP, HP and LDHB had significant change between VTE and non-VTE patients, with the area under the curve (AUC) were 0.8067, 0.8308, 0.7767, 0.8021, 0.8533, respectively. Conclusions SAA1, S100A8, LBP, HP and LDHB may serve as potential plasma biomarkers for diagnosis VTE in NSCLC patients.

Simple Summary The most life-threatening events in colorectal cancer (CRC) are metastasis and thrombosis. Platelets can play a role in these outcomes via the release of medium-sized extracellular vesicles (mEVs). Thus, we aimed to study the EVs released from activated platelets of CRC patients and healthy controls (HS) for their size composition, protein content, and the capacity to influence the expression of genes involved in malignancy and the synthesis of a prothrombotic lipid mediator such as thromboxane (TX)A2. Our findings show that the protein content of thrombin-stimulated mEVs is modulated in CRC. Its evaluation may represent a noninvasive tool to discriminate patients from healthy subjects. Moreover, our findings show that characterizing the regulation of the expression of promalignant genes and prothrombotic phenotypes in cancer cells by the crosstalk with platelet mEVs could provide prognostic information on cancer patients, which could help in developing an appropriate anticancer strategy. Abstract Background: Platelet–cancer cell interactions modulate tumor metastasis and thrombosis in cancer. Platelet-derived extracellular vesicles (EVs) can contribute to these outcomes. Methods: We characterized the medium-sized EVs (mEVs) released by thrombin-stimulated platelets of colorectal cancer (CRC) patients and healthy subjects (HS) on the capacity to induce epithelial-mesenchymal transition (EMT)-related genes and cyclooxygenase (COX)-2(PTGS2), and thromboxane (TX)B2 production in cocultures with four colorectal cancer cell lines. Platelet-derived mEVs were assessed for their size distribution and proteomics signature. Results: The mEV population released from thrombin-activated platelets of CRC patients had a different size distribution vs. HS. Platelet-derived mEVs from CRC patients, but not from HS, upregulated EMT marker genes, such as TWIST1 and VIM, and downregulated CDH1. PTGS2 was also upregulated. In cocultures of platelet-derived mEVs with cancer cells, TXB2 generation was enhanced. The proteomics profile of mEVs released from activated platelets of CRC patients revealed that 119 proteins were downregulated and 89 upregulated vs. HS. Conclusions: We show that mEVs released from thrombin-activated platelets of CRC patients have distinct features (size distribution and proteomics cargo) vs. HS and promote prometastatic and prothrombotic phenotypes in cancer cells. The analysis of platelet-derived mEVs from CRC patients could provide valuable information for developing an appropriate treatment plan.

Introduction Clinical risk assessment scores can help risk-stratify patients for cancer-associated thrombosis (CAT); however, their performance in individual cancer subtype remains suboptimal. While previous studies have explored the utility of single plasma biomarkers such as D-dimer or P-selectin, we lack a validated, non-invasive biomarker signature panel. Our current study evaluates a plasma proteomic assay and machine learning model for predicting CAT. Methods We performed a nested case control study from 1,694 patients with newly diagnosed solid tumor malignancy who had plasma sample collection from 2011-2023 at the Harris Health System. Patients with early-stage disease, samples taken before cancer diagnosis or after chemotherapy initiation, or inadequate follow-up were excluded. Five common cancer types were selected (breast, colorectal, lung, pancreatic, gastroesophageal) to form an eligible cohort of 312 patients. Incidence density sampling was used with 1:2 matching on cancer type, stage, and treatment to form a final analytic cohort of 170 patients (57 CAT vs 113 controls). We performed proteomic profiling using the Olink Explore HT, which uses the proximity extension assay to measure plasma proteins as normalized protein expression (NPX) values. Random Survival Forest (RSF) model was used to predict time to CAT, where NPX values (5,416 proteins) and clinical variables (age, sex, race, ethnicity, cancer type, stage, diagnosis year) were trained in 100 bootstrapped iterations with resample. Internal validation with time dependent receiver operating characteristic (TD-ROC) was assessed at days 30, 90, and 180, using out-of-bag (OOB) test samples. Top proteins identified by nonparametric permutation variable importance (VIMP) and stably present in multiple iterations were retained for pathway enrichment analysis using the Reactome Over-Representation Analysis with p-value <0.05 and FDR q-value <0.1. Results Among 170 patients, the median age was 54 (IQR 47-60), 64% were female. Race and ethnicity included 62% Hispanic, 12% Non-Hispanic (NH) White, 18% NH Black, and 7% NH Asian. There were 42 breast, 51 colorectal, 35 lung, 22 pancreatic, and 20 gastroesophageal cancers; 53% were metastatic; and all received chemotherapy after sample collection. The 57 CAT events included 23 pulmonary embolism, 12 lower extremity deep vein thrombosis (DVT), 18 upper extremity DVT, and 4 splanchnic vein thrombosis. Median time to CAT was 154 days (IQR 70-262). There were 7, 19, and 31 events by 30, 90, and 180 days, respectively. Among 5,416 proteins, the NPX values ranged from -11.43 to +12.40 (IQR -0.50 to +0.52). There were no significant differences by batch or storage time. For internal validation, the RSF model achieved a bootstrapped mean TD-ROC of 0.82 (95% CI 0.54-0.98), 0.71 (95% CI 0.57-0.88), and 0.67 (95% CI 0.56-0.79) at 30, 90, and 180 days, respectively. Approximately 35% were classified as high-risk. In the OOB test sets, the observed average VTE incidence in the high-risk group were 13%, 21%, and 26% at 30d, 90d, and 180d, respectively. In comparison, the observed average VTE incidence in the low-risk group was 1%, 8%, and 15% at the same time points. Top important proteins included KANK1, F9, CABP4, PRND, and GFPT2, among others. In pathway enrichment analyses, the top 20% proteomic features were over-represented in neutrophil degranulation, extracellular matrix organization, platelet cytosolic calcium and degranulation, integrin cell surface interactions, regulation of complement cascade, and formation of fibrin clot. Conclusion Individual cancer patients have unique prothrombotic expression profiles in the plasma that are prognostic for future CAT occurrence. As demonstrated in the pathway enrichment analyses from the top RSF features, the relationship between coagulation cascade, neutrophil, platelet, complement, and extracellular matrix is complex and non-linear. A survival tree-based machine learning model incorporating all plasma proteomic signatures can predict short-term CAT occurrence with higher accuracy than long-term (0.82 vs 0.67). Instead of single target assays, panel-based plasma proteomic assays may complement existing clinical risk scores and further differentiate the thrombotic risk profiles among individual cancer subtypes. External and prospective cohort validations are needed to ensure reproducibility and generalizability.

Background Cancer patients have an increased risk of developing venous thromboembolism, with up to 30% dying within a month of their development. Some cancer cells are known to induce platelet aggregation, and this interaction is understood to contribute to thrombosis and haematogenous metastasis. Many researchers have reported on extracellular vesicles (EVs) released from platelets. However, less is known about how cancer cells’ EVs may affect platelet function. Here EVs released by triple-negative breast cancer (TNBC) cell line variants were extensively investigated in this regard. Methods EVs were separated from conditioned media of TNBC Hs578T and Hs578Ts(i) 8 cells using filtration and ultracentrifugation and were characterised by nanoparticle tracking analysis, immunoblots, and transmission electron microscopy. Blood samples from consenting donors were procured, and their platelets collected by differential centrifugation. Light transmission aggregometry and optical microscopy evaluated the potential interaction of TNBC cells and their EVs with platelets. Global proteomic analysis was performed on the EVs, by in-solution digestion and mass spectrometry. Data analysis included the use of Perseus, FunRich, and Vesiclepedia. Immunoblotting was used as a secondary method to investigate some key EV cargo proteins identified by the global proteomics approach. Results Both TNBC cell variants induced platelet aggregation. Increasing cell numbers significantly reduced the time taken for platelet aggregation to occur. EVs released by the cells also resulted in platelet aggregation. The time to induce platelet aggregation was EV dose-dependent. Proteomics profiling and immunoblotting of the EVs’ cargo identified candidate proteins (including uPAR and PDGFRβ) that may be involved during this process. Conclusions TNBC cells induce platelet aggregation. Furthermore, the cell-free EVs induced this undesirable effect. A number of EV cargo proteins were identified that may be relevant as therapeutic targets.

Pancreatic cancer\nis the third leading cause of cancer-related\ndeath in the USA. Despite extensive research, minimal improvements\nin patient outcomes have been achieved. Early identification of treatment\nresponse and metastasis would be valuable to determine the appropriate\ntherapeutic course for patients. In this work, we isolated exosomes\nfrom the serum of 10 patients with locally advanced pancreatic cancer\nat serial time points over a course of therapy, and quantitative analysis\nwas performed using the iTRAQ method. We detected approximately 700–800\nexosomal proteins per sample, several of which have been implicated\nin metastasis and treatment resistance. We compared the exosomal proteome\nof patients at different time points during treatment to healthy controls\nand identified eight proteins that show global treatment-specific\nchanges. We then tested the effect of patient-derived exosomes on\nthe migration of tumor cells and found that patient-derived exosomes,\nbut not healthy controls, induce cell migration, supporting their\nrole in metastasis. Our data show that exosomes can be reliably extracted\nfrom patient serum and analyzed for protein content. The differential\nloading of exosomes during a course of therapy suggests that exosomes\nmay provide novel insights into the development of treatment resistance\nand metastasis.

… cancers and normal controls, and to develop useful tumor markers of gastric cancer by quantitative proteomic … of patients in an advanced cancer group, early cancer group, and normal …

Key Points • We applied proteomic profiling to patients with suspected HIT, thus analyzing a large number of potential proteins.• Our analysis provided evidence supporting the potential of soluble P-selectin as a promising new biomarker in HIT.

Background Venous thromboembolism (VTE) has been associated with several modifiable factors (MFs) and cardiovascular comorbidities. However, the mechanisms are largely unknown. Objectives We aimed to decipher proteomic pathways underlying the associations of VTE with MFs and cardiovascular comorbidities. Methods A 2-stage network Mendelian randomization analysis was conducted to explore the associations between 15 MFs, 1151 blood proteins, and VTE using data from a genome-wide meta-analysis including 81 190 cases of VTE. We used protein data from 35 559 individuals as the discovery analysis, and from 2 independent studies including 10 708 and 54 219 participants as the replication analyses. Based on the identified proteins, we assessed the druggability and examined the cardiovascular pleiotropy. Results The network Mendelian randomization analyses identified 10 MF–VTE, 86 MF–protein, and 34 protein–VTE associations. These associations were overall consistent in the replication analyses. Thirty-eight pathways with directionally consistent direct and indirect effects in the MF–protein–VTE pathway were identified. Low-density lipoprotein receptor–related protein 12 (LRP12: 34.3%-58.1%) and coagulation factor (F)XI (20.6%-39.6%) mediated most of the associations between 3 obesity indicators and VTE. Likewise, coagulation FXI mediated most of the smoking–VTE association (40%; 95% CI, 20%-60%) and insomnia–VTE association (27%; 95% CI, 5%-49%). Many VTE-associated proteins were highly druggable for thrombotic conditions. Five proteins (interleukin-6 receptor subunit alpha, LRP12, prothrombin, angiopoietin-1, and low-density lipoprotein receptor–related protein 4) were associated with VTE and its cardiovascular comorbidities. Conclusion This study suggests that coagulation FXI, a druggable target, is an important mediator of the associations of obesity, smoking, and insomnia with VTE risk.

… thrombus into the pulmonary artery, which has a mortality rate of up to 30% [3]. We developed a proteomic … or serum as a primary source of proteomic biomarkers, such as its accessibility…

It has been reported that 476 proteins can be detected in plasma fibrin clots from patients with venous thromboembolism. Plasma fibrin clots proteomic composition in relation to their properties has not been studied in acute pulmonary embolism (PE). Clots generated from plasma of 20 PE patients and 20 healthy controls were assessed using mass spectrometry, clot permeability (Ks), and clot lysis time (CLT). The proteomic composition of plasma fibrin clots from acute PE patients differed from that of control subjects in regard to 198 clot-bound proteins. In the acute PE group, we observed increased clot-bound fibrinogen, apolipoprotein B-100, platelet glycoprotein Ib, lipopolysaccharide-binding protein, and histones H3 + 4 and reduced fibronectin, α2-antiplasmin, α2-macroglobulin, factor (F)XIII, histidine-rich glycoprotein, antithrombin, von Willebrand Factor, plasminogen, and prothrombin. Among PE patients, low Ks (≤3.83 × 10-9 cm2) was associated with increased clot-bound C-reactive protein, kininogen-1, protein S, β-2-microglobulin, and thromboxane-A synthase when compared with patients having Ks > 3.83 × 10-9 cm2. Ks correlated inversely with FIX and FV, thrombin-activatable fibrinolysis inhibitor, complement C1s, C7, C8, and apolipoprotein A-I. The specific protein composition in plasma fibrin clots from acute PE patients is associated with denser clot formation. Several proteins unrelated to the coagulation system can modulate fibrin phenotype in acute thrombotic states. SIGNIFICANCE: Our study significantly advances the field of thrombosis and hemostasis. The plasma fibrin clot proteomics findings fill the gap of knowledge about the presence and the role of other proteins to the plasma fibrin clot in the acute phase of pulmonary embolism, aside fibrinogen, which is the main component of fibrin. The reported methodology, which involves the sample preparation using Multienzyme Digestion-Filter Aided Sample Preparation (MED FASP), data acquisition with the Quadrupole-Orbitrap mass spectrometer, and data analysis using the advanced tools such as MaxQuant, Total Protein Approach and Perseus, allows to gain not only the qualitative, but also the quantitative insights into the microworld of proteins entangled among the fibrin network. By comparing the clots formed from plasma of patients with acute pulmonary embolism with the clots from healthy control, we provide the specific protein composition associated with unfavorable clot properties observed in this disease. Moreover, our findings emphasize that several proteins unrelated to the coagulation system, can modulate fibrin phenotype in acute thrombotic states.

Cancer is associated with a systemic hypercoagulable state and an increased risk of acute ischemic stroke (AIS), yet the mechanisms underlying cancer-associated arterial thrombosis remain poorly defined. We conducted comprehensive analyses of blood and thrombi in patients with AIS and Cancer (AIS-Cancer), AIS without cancer, and cancer without AIS. AIS-Cancer showed the highest neutrophil activation, whereas biomarkers of systemic inflammation, endothelial dysfunction, and peripheral platelet phenotypes (proaggregant and procoagulant platelets and AV+ platelet-derived microvesicles) were similar across groups. In contrast, thrombi retrieved from large vessels in AIS-Cancer were predominantly platelet-rich and contained fibrin, neutrophils and neutrophil extracellular traps, whereas thrombi from non-cancer AIS were mainly red blood cell-rich. Platelets within AIS-Cancer thrombi exhibited a distinct procoagulant phenotype elicited in vitro with plasma from affected patients. Together, these findings suggest that cancer reprograms arterial thrombus formation toward a procoagulant and NET-rich phenotype, providing a rationale for therapies targeting procoagulant platelet activity in cancer-associated stroke.

… Collectively, platelet-derived mitochondrial transfer represents an emerging layer of intercellular communication that may link thrombosis, metabolism, and metastasis, and it offers …

Several studies have suggested a role for blood coagulation proteins in tumour progression. Herein, we discuss (1) the activation of the blood clotting cascade in the tumour microenvironment and its impact on primary tumour growth; (2) the intravascular activation of blood coagulation and its impact on tumour metastasis and cancer-associated thrombosis; and (3) antitumour therapies that target blood-coagulation-associated proteins. Expression levels of the clotting initiator protein TF (tissue factor) have been correlated with tumour cell aggressiveness. Simultaneous TF expression and PS (phosphatidylserine) exposure by tumour cells promote the extravascular activation of blood coagulation. The generation of blood coagulation enzymes in the tumour microenvironment may trigger the activation of PARs (protease-activated receptors). In particular, PAR1 and PAR2 have been associated with many aspects of tumour biology. The procoagulant activity of circulating tumour cells favours metastasis, whereas the release of TF-bearing MVs (microvesicles) into the circulation has been correlated with cancer-associated thrombosis. Given the role of coagulation proteins in tumour progression, it has been proposed that they could be targets for the development of new antitumour therapies.

… We believe that TF contributes to tumor angiogenesis via both clotting-… cycle of tumor growth and clot formation. VEGF also increases vascular permeability that leads to plasma protein …

Cancer is associated with an increased risk of venous thromboembolism (VTE); the exact mechanisms for the induction of VTE remain to be fully elucidated, but it is widely acknowledged that tissue factor (TF)‐bearing microparticles (TF‐MPs) may play a significant role. However, TF‐MPs have yet to be accepted as a genuine biomarker for cancer‐associated VTE, as the presence of elevated TF‐MP levels is not always accompanied by thrombosis; interestingly, in certain cases, particularly in pancreatic cancer, VTE seems to be more likely in the context of acute inflammation. Although several potential mechanisms for the development of VTE in cancer have been postulated, this review explores the homeostatic disruption of TF‐MPs, as the main reservoir of bloodborne TF, in the context of cancer and inflammation, and considers the abrogated responses of the activated endothelium and mononuclear phagocyte system in mediating this disruption.

Blood coagulation appears to play an important role in the occurrence of cancer and its effects may be twofold. First, in patients with cancer, blood coagulation is activated in the direction of a prothrombotic state. Second, a procoagulant environment may promote cancer in different ways. In this chapter we discuss some of the mechanisms that may be involved in this interplay between coagulation and cancer. Blood coagulation proteins interact with cells in the vasculature to maintain hemostasis. However, many proteins that are involved in coagulation and anticoagulation, as well as fibrinolysis, are also found in extravascular tissues. In different organs, these proteins may be involved in cell-signaling mechanisms, through interaction with cell receptors like protease-activated receptors (PARs). Such interactions may drive inflammation, angiogenesis and cell proliferation. The potential procarcinogenic actions of proteases like thrombin may be counteracted by the anticoagulant and anti-inflammatory actions of the protein C-thrombomodulin mechanism. In the blood of cancer patients, the balance is usually shifted towards a procoagulant direction. The resulting excess thrombin- and fibrin-forming activity promotes venous thrombosis and may in the extravascular compartment stimulate cancer progression. The activation of platelets and their interaction with leukocytes may propagate this process. In addition to the therapeutic modulation of the prothrombotic environment, the induction of specific anticoagulant proteins including thrombomodulin may have effects on tumor growth or dissemination, but the nature of these effects still remains hard to predict. The interplay between cancer and blood coagulation merits further experimental and clinical research.

A prothrombotic state is frequently observed in patients with cancer and contributes to the risks of venous thromboembolism (VTE), arterial thromboembolism (ATE), tumor progression, and death. Altered ex vivo properties of plasma clot formation and lysis have been observed in patients with cancer. The aim of this prospective study was to comprehensively characterize the relationship between plasma clot properties, inflammation, hypercoagulability, thrombotic complications, and mortality in patients with cancer using a tissue-factor-based turbidimetric assay of clot formation and lysis. Turbidity parameters were determined in 815 patients with newly-diagnosed or recurrent cancer and 97 healthy controls. Patients were followed-up for two years and rates of VTE (n=72 events), ATE (n=21 events), and death (n=304 events) were assessed. Compared to controls, cancer patients’ turbidity profiles showed an increased clot formation potential and higher resistance towards fibrinolysis. Elevated biomarkers of inflammation and hemostasis, such as C-reactive protein, FVIII, and thrombin generation explained substantial amounts of variation in turbidity parameters. In a prospective analysis, altered parameters of clot formation identified cancer patients at high risk of ATE (Hazard ratio (HR) per doubling of peak absorbance: 4.43, 95% CI: 1.50-13.07, p=0.007) and death (HR per doubling of peak absorbance: 2.73, 2.00-3.72, p<0.0001); these findings were independent of other prognostic covariates. Contrarily, turbidity parameters were not associated with risk of VTE (HR per doubling of peak absorbance: 1.15, 0.66-2.01, p=0.62). We conclude that patients with cancer have altered ex vivo properties of clot formation which predict risks of ATE and mortality but not VTE.

… It is important to note however that, overall, breast cancer is associated with a relatively low thrombosis risk, and also that the chemotherapy in this study was administered as an …

… patients with localized cancer [4]… thrombosis may be the first sign of a malignant tumor [5]. Finally, activation of the coagulation cascade and aggregation of blood platelets around cancer …

… This study shows that a positive TpP level is a predictor of tumor thrombosis in HCC, which … between plasma D-dimer and TpP levels with tumor thrombosis. Positive plasma levels of …

… both thrombogenesis and tumor progression. Besides … tumor, and anticancer therapies increase the thrombotic risk in these patients. Furthermore, biological factors, including the cancer …

… Such results lead to a hypothesis that tumor cells may produce soluble sub… of tumors permit tumor products to enter the systemic circulation continuously and/or allow plasma clotting …

Simple Summary The interactions between circulating tumor cells (CTCs) and plasma proteins are critical for hematogenous metastasis, and understanding the molecular mechanisms behind these interactions can improve liquid-biopsy-based diagnostics and cancer therapies. This review summarizes recent literature on the surface molecules of CTCs that interact with coagulation proteins and their biological and clinical relevance. It also discusses future research directions to expand our knowledge of the CTC interactome, which can lead to the discovery of new molecular markers for diagnostics and additional targets for cancer therapies. Abstract Cancer metastasis is a complex process. After their intravasation into the circulation, the cancer cells are exposed to a harsh environment of physical and biochemical hazards. Whether circulating tumor cells (CTCs) survive and escape from blood flow defines their ability to metastasize. CTCs sense their environment with surface-exposed receptors. The recognition of corresponding ligands, e.g., fibrinogen, by integrins can induce intracellular signaling processes driving CTCs’ survival. Other receptors, such as tissue factor (TF), enable CTCs to induce coagulation. Cancer-associated thrombosis (CAT) is adversely connected to patients’ outcome. However, cancer cells have also the ability to inhibit coagulation, e.g., through expressing thrombomodulin (TM) or heparan sulfate (HS), an activator of antithrombin (AT). To that extent, individual CTCs can interact with plasma proteins, and whether these interactions are connected to metastasis or clinical symptoms such as CAT is largely unknown. In the present review, we discuss the biological and clinical relevance of cancer-cell-expressed surface molecules and their interaction with plasma proteins. We aim to encourage future research to expand our knowledge of the CTC interactome, as this may not only yield new molecular markers improving liquid-biopsy-based diagnostics but also additional targets for better cancer therapies.

We have previously proposed that intravascular thrombosis and subsequent vasoocclusion contribute to the development of pseudopalisading necrosis, a pathologic hallmark that distinguishes glioblastoma (WHO grade 4) from lower grade astrocytomas. To better understand the potential prothrombotic mechanisms underlying the formation of these structures that drive tumor angiogenesis, we investigated tissue factor (TF), a potent procoagulant protein known to be overexpressed in astrocytomas. We hypothesized that PTEN loss and tumor hypoxia, which characterize glioblastoma but not lower grade astrocytomas, could up-regulate TF expression and cause intravascular thrombotic occlusion. We examined the effect of PTEN restoration and hypoxia on TF expression and plasma coagulation using a human glioma cell line containing an inducible wt-PTEN cDNA. Cell exposure to hypoxia (1% O(2)) markedly increased TF expression, whereas restoration of wt-PTEN caused decreased cellular TF. The latter effect was at least partially dependent on PTEN's protein phosphatase activity. Hypoxic cells accelerated plasma clotting in tilt tube assays and this effect was prevented by both inhibitory antibodies to TF and plasma lacking factor VII, implicating TF-dependent mechanisms. To further examine the genetic events leading to TF up-regulation during progression of astrocytomas, we investigated its expression in a series of human astrocytes sequentially infected with E6/E7/human telomerase, Ras, and Akt. Cells transformed with Akt showed the greatest incremental increase in hypoxia-induced TF expression and secretion. Together, our results show that PTEN loss and hypoxia up-regulate TF expression and promote plasma clotting by glioma cells, suggesting that these mechanisms may underlie intravascular thrombosis and pseudopalisading necrosis in glioblastoma.

… As a result, a microenvironment promoting tumor growth is … cancer-thrombosis connection exists, by which cancer cells support clot formation, and clotting proteins support cancer …

… For the measurement of MP-associated TF activity (MP-TF activity), MP were washed more extensively to reduce contamination with plasma proteins (0.5% in the final MP preparation) …

Tissue factor (TF) is the primary initiator of the coagulation cascade, though its effects extend well beyond hemostasis. When TF binds to Factor VII, the resulting TF:FVIIa complex can proteolytically cleave transmembrane G protein-coupled protease-activated receptors (PARs). In addition to activating PARs, TF:FVIIa complex can also activate receptor tyrosine kinases (RTKs) and integrins. These signaling pathways are utilized by tumors to increase cell proliferation, angiogenesis, metastasis, and cancer stem-like cell maintenance. Herein, we review in detail the regulation of TF expression, mechanisms of TF signaling, their pathological consequences, and how it is being targeted in experimental cancer therapeutics.

… These and other accompanying changes in the structure of the vessel wall trigger vascular permeability, extravasation of plasma proteins, microhemorrhage, extravascular clotting, and …

Venous and arterial thromboses, called as cancer-associated thromboembolism (CAT), are common complications in cancer patients that are associated with high mortality. The cell-surface glycoprotein tissue factor (TF) initiates the extrinsic blood coagulation cascade. TF is overexpressed in cancer cells and is a component of extracellular vesicles (EVs). Shedding of TF+EVs from cancer cells followed by association with coagulation factor VII (fVII) can trigger the blood coagulation cascade, followed by cancer-associated venous thromboembolism in some cancer types. Secretion of TF is controlled by multiple mechanisms of TF+EV biogenesis. The procoagulant function of TF is regulated via its conformational change. Thus, multiple steps participate in the elevation of plasma procoagulant activity. Whether cancer cell-derived TF is maximally active in the blood is unclear. Numerous mechanisms other than TF+EVs have been proposed as possible causes of CAT. In this review, we focused on a wide variety of regulatory and shedding mechanisms for TF, including the effect of SARS-CoV-2, to provide a broad overview for its role in CAT. Furthermore, we present the current technical issues in studying the relationship between CAT and TF.

… Background: Tissue factor (TF)-bearing microparticles (MP) from different origins are … in the pathogenesis of cancer-associated thrombosis. However, the role of circulating tumor cell-…

Tumours are capable of activating blood coagulation through the expression of procoagulant molecules such as tissue factor, cancer procoagulant and hepsin. Initiation of the clotting cascade results in the generation of the activated serine proteases factor VIIa, factor Xa and thrombin. These proteases act via protease-activated receptors and tissue factor to alter gene expression, thereby modulating tumour cell growth, invasion, metastasis and angiogenesis.

… to stimulate synthesis of coagulation initiators by host endothelial cells or monocytes/macrophages. Such an indirect mechanism may explain systemic coagulation activation in the type …

Routine coagulation tests are not widely accepted diagnostic criteria of trauma-induced hypercoagulopathy (TIH) due to insensitivity. Lymphatic vessels drain approximately 10% of the interstitial fluid into the lymphatic system and form lymph. The purpose of this study was to identify the potential lymph biomarkers for TIH. Eighteen male Sprague-Dawley rats were randomly assigned to the sham (non-fractured rats with sham surgery and vehicle treatment), the VEH (fractured rats with vehicle treatment) and the CLO (fractured rats with clopidogrel treatment) group. Thoracic duct lymph was obtained to perform proteomics and untargeted metabolomics. A total of 1207 proteins and 16,695 metabolites were identified. The top 5 GO terms of lymph proteomics indicated that oxidative stress and innate immunity were closely associated with TIH and antithrombotic therapy. The top 5 GO terms of lymph metabolomics showed that homocystine and lysophosphatidylcholine were the differential expressed metabolites (DEMs) between the sham and VEH groups, while cholic acid, docosahexaenoic acid, N1-Methyl-2-pyridone-5-carboxamide, isoleucine and testosterone are the DEMs between the VEH and CLO group. This study presents the first proteomic and metabolomic profiling of lymph after TIH and antithrombotic therapy, and predicts the possible lymph biomarkers for TIH.

Background Venous thromboembolism (VTE) is a complex thrombotic disorder that constitutes a major source of mortality and morbidity. To improve understanding of the cause of VTE, we conducted a metabolomic analysis in a case‐control study including 240 incident VTE cases and 6963 controls nested within 3 large prospective population‐based cohorts, the Nurses’ Health Study, the Nurses’ Health Study II, and the Health Professionals Follow‐Up Study. Methods and Results For each individual, we measured 211 metabolites and collected detailed information on lifestyle factors. We performed logistic regression and enrichment analysis to identify metabolites and biological categories associated with incident VTE risk, accounting for key confounders, such as age, sex, smoking, alcohol consumption, body mass index, and comorbid diseases (eg, cancers). We performed analyses of all VTEs and separate analyses of pulmonary embolism. Using the basic model controlling for age, sex, and primary disease, we identified 60 nominally significant VTE‐ or pulmonary embolism–associated metabolites (P<0.05). These metabolites were enriched for diacylglycerols (P permutation<0.05). However, after controlling for multiple testing, only 1 metabolite (C5 carnitine; odds ratio, 1.25; 95% confidence interval, 1.10–1.41; P corrected=0.03) remained significantly associated with VTE. After further adjustment for body mass index, no metabolites were significantly associated with disease after accounting for multiple testing, and no metabolite classes were enriched for nominally significant associations. Conclusions Although our findings suggest that circulating metabolites may influence the risk of incident VTE, the associations we observed were confounded by body mass index. Larger studies involving additional individuals and with broader metabolomics coverage are needed to confirm our findings.

Deep vein thrombosis (DVT) is a serious health issue that often leads to considerable morbidity and mortality. Diagnosis of DVT in a clinical setting, however, presents considerable challenges. The fusion of metabolomics techniques and machine learning methods has led to high diagnostic and prognostic accuracy for various pathological conditions. This study explored the synergistic potential of dual-platform metabolomics (specifically, gas chromatography–mass spectrometry (GC-MS) and liquid chromatography–mass spectrometry (LC-MS)) to expand the detection of metabolites and improve the precision of DVT diagnosis. Sixty-one differential metabolites were identified in serum from DVT patients: 22 from GC-MS and 39 from LC-MS. Among these, five key metabolites were highlighted by SHapley Additive exPlanations (SHAP)-guided feature engineering and then used to develop a stacking diagnostic model. Additionally, a user-friendly interface application system was developed to streamline and automate the application of the diagnostic model, enhancing its practicality and accessibility for clinical use. This work showed that the integration of dual-platform metabolomics with a stacking machine learning model enables faster and more accurate diagnosis of DVT in clinical environments.

In recent years, the field of venous thromboembolism has undergone numerous innovations, starting from the recent discoveries on the role of biomarkers, passing through the role of metabolomics in expanding our knowledge on pathogenic mechanisms, which have opened up new therapeutic targets. A variety of studies have contributed to characterizing the metabolic phenotype that occurs in venous thromboembolism, identifying numerous pathways that are altered in this setting. Among these pathways are the metabolism of carnitine, tryptophan, purine, and fatty acids. Furthermore, new evidence has emerged with the recent COVID-19 pandemic. Hypercoagulability phenomena induced by this viral infection appear to be related to altered von Willebrand factor activity, alteration of the renin–angiotensin–aldosterone system, and dysregulation of both innate and adaptive immunity. This is the first literature review that brings together the most recent evidence regarding biomarkers, metabolomics, and COVID-19 in the field of venous thromboembolism, while also mentioning current therapeutic protocols.

Deep vein thrombosis (DVT) is a significant contributor to cardiovascular morbidity and mortality, which can be classified as muscular calf vein thrombosis (MCVT) or popliteal vein thrombosis (PVTE). This study aimed to evaluate the differential metabolites of DVT using ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Metabolic profiles were analyzed, and binary logistic regression was used to build a diagnostic model to identify possible potential biomarkers for distinguishing DVT and its subtypes. The area under the curve (AUC) of the receiver operating characteristic (ROC) curves was employed to evaluate the diagnostic performance of these possible potential biomarkers. In this study, eight, three, and six differential metabolites associated with DVT, MCVT, and PVTE were identified. Enrichment analysis of differential metabolites revealed that DVT was mainly associated with energy metabolism, purine metabolism, and amino acid metabolism. Diagnostic models were developed based on differential metabolites; the diagnostic performance of DVT, MCVT, and PVTE was excellent. Our findings revealed distinct metabolic profiles for DVT, MCVT, and PVTE. Despite the small sample size and the inclusion of only Asian populations, this study paves the way for future large-sample multicenter follow-up studies.

Dipsaci Radix may possess antithrombotic properties, and one of its primary active ingredients is Asperosaponin VI. However, the antithrombotic effects and pharmacological mechanisms of Asperosaponin VI remain unclear. An in vivo experimental study has demonstrated the antithrombotic activity of Asperosaponin VI. Asperosaponin VI also exhibits anticoagulant properties. Asperosaponin VI significantly hindered collagen adrenergic-induced acute pulmonary thrombosis in mice and enhanced their survival rate. This hinders the formation of acute pulmonary embolisms induced by adenosine diphosphate (ADP) and decreases recovery time. A comprehensive strategy that combines metabolomics, network pharmacology, molecular docking, and experimental validation has the potential to reveal the antithrombotic mechanisms of Asperosaponin VI. Metabolomic evidence suggests that Asperosaponin VI may influence platelet aggregation and the production of anti-inflammatory metabolites through the regulation of pathways such as phenylalanine and arachidonic acid metabolism, thereby inhibiting thrombosis. Network pharmacology identified the pharmacological targets of Asperosaponin VI and indicated that it treats thrombi by partially regulating the signaling pathways related to inflammation and platelet aggregation. Asperosaponin VI showed strong binding affinity for F2, PTPRC, JUN, STAT3, SRC, AKT1. The antiplatelet aggregation activity of Asperosaponin VI was validated based on the metabolomic and network pharmacology results. Asperosaponin VI inhibits platelet aggregation induced by ADP, AA, and collagen. Therefore, Asperosaponin VI exerts antithrombotic effects through antiplatelet aggregation. Therefore, Asperosaponin VI is a promising antithrombotic agent.

Deep vein thrombosis (DVT) of the lower extremities is one of the most common peripheral vascular diseases, with significant complications and sequelae. Metabolomics aims to identify small molecules in biological samples. It can serve as a promising method for screening compounds that can be used for early disease detection, diagnosis, treatment response prediction, and prognosis. In addition, high-throughput metabolomics screening can yield significant insights into the pathophysiological pathways of DVT. Currently, the metabolomic profiles of DVT have yielded inconsistent expression patterns. This article examines the recent advancements in metabolomic studies of DVT and analyzes the factors that may influence the results.

Deep vein thrombosis (DVT) is a leading cause of morbidity and mortality after trauma. Here, we integrate plasma metabolomics and proteomics to evaluate the metabolic alterations and their function in up to 680 individuals with and without DVT after trauma (pt-DVT). We identify 28 metabolites and 2 clinical parameter clusters associated with pt-DVT. Then, we develop a panel of 9 metabolites (hexadecanedioic acid, pyruvic acid, L-Carnitine, serotonin, PE(P-18:1(11Z)/18:2(9Z,12Z)), 3-Hydroxycapric acid, 5,6-DHET, 3-Methoxybenzenepropanoic acid and pentanenitrile) that can predict pt-DVT with high performance, which can be verified in an independent cohort. Furthermore, the integration analysis of metabolomics and proteomics data indicates that the upregulation of glycolysis/gluconeogenesis-TCA cycle may promote thrombosis by regulating ROS levels in red blood cells, suggesting that interfering with this process might be potential therapeutic strategies for pt-DVT. Together, our study comprehensively delineates the metabolic and hematological dysregulations for pt-DVT, and provides potential biomarkers for early detection. Deep vein thrombosis after trauma (pt-DVT) patients undergo metabolic changes but a comprehensive characterization in humans is lacking. Here the authors integrate metabolome and proteome to reveal pt-DVT-related metabolic alterations and their function, and reveal potential biomarkers.

Abstract Thrombosis is a relatively common human disease, being the third cause of mortality worldwide. Alterations in hemostasis, endothelial processes and blood flow are the major features involved in the pathophysiology of the disease, whose molecular basis is not yet fully understood due to the complexity and numerous influences that might lead to blood clotting. The activity and the role of blood metabolites in thrombosis are attracting great attention in the quest of understanding the triggering off uncontrolled clotting. Metabolomics can bring insights into metabolic fingerprints of any biological sample, quantify, and measure the metabolic responses to the pathophysiological stimuli or genetic modifications, and lead to an integrated comprehension of metabolites function in health and disease. Furthermore, the identification of biomarkers can reveal metabolic pathways associated with the pathogenesis and progression of the disease. In this review, we summarized the current state of knowledge of metabolomics in arterial and venous thrombosis research by nuclear magnetic resonance spectroscopy and mass spectrometry approaches and cited and discussed the altered metabolites that have been cited in both conditions. It is worth stating that the contribution of the metabolomics in this field or research is modest and being introduced in some case studies, and expected to grow in the near future.

Essentials Risk‐stratification often fails to predict clinical deterioration in pulmonary embolism (PE). First‐ever high‐throughput metabolomics analysis of risk‐stratified PE patients. Changes in circulating metabolites reflect a compromised energy metabolism in PE. Metabolites play a key role in the pathophysiology and risk stratification of PE.

Objective This study aims to integrate metabolomics and transcriptomics data to investigate the protective effects of umbilical cord mesenchymal stem cells (UC-MSCs) on obstetric deep vein thrombosis (DVT) and to elucidate the underlying molecular mechanisms. Methods A pregnant rat model of DVT was established using the inferior vena cava (IVC) stenosis method. The protective effects of UC-MSCs on DVT and endothelial cell injury were evaluated both in vivo and in vitro. Transcriptomic and metabolomic analyses were performed to identify differentially expressed genes (DEGs) and differentially abundant metabolites (DMs) in IVC tissues from DVT rats and those treated with UC-MSCs. Correlation analysis was conducted to associate relevant metabolites and RNAs. Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was applied to DEGs and DMs to identify significantly involved pathways. The mRNA–transcription factor regulatory network was constructed using Cytoscape software. Receiver operating characteristic (ROC) curves for immune regulatory genes and DEGs were generated with the R package pROC. The mMCP-counter algorithm was used to assess the distribution and abundance of immune cell subsets. Results The rat DVT model was established using the IVC stenosis method. Administration of UC-MSCs reduced thrombus burden, promoted angiogenesis, and mitigated hydrogen peroxide-induced endothelial injury in the DVT model. Integrated transcriptomic and metabolomic analyses revealed significant correlations between four key metabolites—pyridine, nicotinamide, L-phenylalanine, and L-leucine—and 24 interacting genes. These metabolites served as critical nodes within the regulatory network. KEGG enrichment analysis indicated that pathways such as amino acid biosynthesis and phenylalanine metabolism are implicated in the therapeutic effects of UC-MSCs on pregnancy-related DVT. Notably, the hub gene Got2 was associated with amino acid biosynthesis, while both Got2 and Maoa were involved in phenylalanine metabolism. Furthermore, seven immune-regulatory genes, including Gaa and Tlr2, demonstrated significant classification performance (area under the curve [AUC] > 0.8) in ROC curve analysis. Conclusion This study elucidates the protective mechanisms of UC-MSCs in the treatment of DVT in pregnant rats induced by the inferior vena cava stenosis model. These findings provide a scientific basis for the further evaluation and development of UC-MSCs-based therapeutic strategies for DVT during pregnancy.

The high occurrence of cancer-associated thrombosis is associated with elevated thrombin generation. Tumour cells increase the potential for thrombin generation both directly, through the expression and release of procoagulant factors, and indirectly, through signals that activate other cell types (including platelets, leukocytes and erythrocytes). Furthermore, cancer treatments can worsen these effects. Coagulation factors, including tissue factor, and inhibitors of coagulation are altered and extracellular vesicles (EVs), which can promote and support thrombin generation, are released by tumour and other cells. Some phosphatidylserine-expressing platelet subsets and platelet-derived EVs provide the surface required for the assembly of coagulation factors essential for thrombin generation in vivo. This review will explore the causes of increased thrombin production in cancer, and the availability and utility of tests and biomarkers. Increased thrombin production not only increases blood coagulation, but also promotes tumour growth and metastasis and as a consequence, thrombin and its contributors present opportunities for treatment of cancer-associated thrombosis and cancer itself.

… cancer and the high incidence of VTE in this population, we hypothesized that measurement of thrombin generation could help stratify patients with cancer … thrombin generation with a …

The association of idiopathic venous thrombosis with occult cancer is generally recognized. However, it has not been fully appreciated that thrombin generated during thrombosis can augment the malignant phenotype. Thrombin activates tumor cell adhesion to platelets, endothelial cells, and subendothelial matrix proteins; enhances tumor cell growth; increases tumor cell seeding and spontaneous metastasis; and stimulates tumor cell angiogenesis. These mechanisms are reviewed. Evidence is also presented to support the hypothesis that thrombin serves to preserve dormant tumor cells in individuals, preventing host eradication. It is proposed that tumor malignancy may be regulated by a procoagulant/anticoagulant axis.

… Advanced cancer is associated with a hypercoagulable state that is triggered by … thrombin generation is crucial for metastasis through fibrin and platelet deposition, as well as thrombin-…

BACKGROUND Thrombin generation, thrombin-antithrombin complex (TAT) levels, and prothrombin fragment 1+2 (F1+2) have shown potential as biomarkers of thromboembolic risk. The aims were to establish reference intervals for these three biomarkers and to assess the levels in patients with localized cancer compared with healthy individuals. METHODS We included 124 healthy individuals (57 females and 67 males; aged 21-66 years), 86 patients with low-stage primary lung cancer, and 57 patients with localized head and neck cancer. Thrombin generation was determined by the calibrated automated thrombogram using platelet-poor-plasma reagent containing 1 pM tissue factor and 4 μM phospholipids. TAT and F1+2 were measured using commercial enzyme-linked immunosorbent assays. Reference intervals were calculated as mean ± 1.96 × standard deviation (thrombin generation and F1+2) or 2.5th to 97.5th percentiles (TAT). RESULTS The reference intervals for thrombin generation parameters were: lag time 4.4-9.4 min, peak thrombin 46-288 nM, time-to-peak thrombin 8-15 min, and endogenous thrombin potential 554-1952 nM x min. The reference interval for TAT was ≤13 μg/l, and for F1+2 it was 47-320 pmol/l. Both low-stage primary lung cancer and head and neck cancer patients had significantly higher TAT (p <0.0001) and F1+2 (p < 0.0001) concentrations than healthy individuals. However, this was not reflected in the thrombin generation assay. CONCLUSION Reference intervals for thrombin generation, TAT as well as F1+2 were established. Patients with localized cancer had significantly elevated TAT and F1+2. TAT and F1+2 may hold potential for identifying hypercoagulation in cancer patients.

… thrombosis is thrombin generation and fibrin formation. The thrombin in circulation is rapidly inactivated by anti-thrombin… for the anti-thrombin therapy of cancer. These events can be …

… We report that thrombin generation and other coagulation parameters are elevated in cachectic but not in non-cachectic tumor-bearing animals, potentially mediated through increased …

INTRODUCTION Thromboembolic disease is a major complication in patients with pancreatic ductal adenocarcinoma (PDAC). Patients with PDAC often have altered blood cell counts, which associate with venous thromboembolism (VTE) development. The high thrombotic risk in patients with PDAC may be partially caused by pro-coagulant blood cells. METHODS The aim of this study was to compare blood-cell dependent coagulation between patients with PDAC (n=18) and healthy controls matched for age and sex (n=18). Thrombin generation (TG) was measured in whole blood (WB) and plasma. The capacity of platelets to release granules (PGRC) was measured in WB. We explored the occurrence of thromboembolic events in PDAC patients during 6-months follow-up. RESULTS Patients showed an increased endogenous thrombin potential (ETP) in WB, compared to controls. This difference was not observed in plasma, indicating a procoagulant effect of blood cells. Both in WB and plasma, the lag time was prolonged in patients compared to controls. Patients had hyperresponsive platelets, with a shorter time to peak granule release. Of the 18 PDAC patients, four developed a VTE (22%) and one an ATE (6%). A shorter lag time in whole blood, not in plasma, and an increased PGRC were associated with thromboembolic events. CONCLUSIONS Patients with PDAC have an increased and delayed WB-TG coagulation profile compared to controls. A shorter lag time in WB-TG and increased PGRC associated with the incidence of thromboembolic events. Platelets appear to be key players in thrombosis development. Measuring hemostasis in whole blood could improve thrombosis risk estimation in PDAC patients.

Patients with localized non-small cell lung cancer (NSCLC) considered unfit for surgery are at substantially increased risk of venous thromboembolism. Radiotherapy may further increase this risk. We aim to investigate the impact of stereotactic body radiotherapy (SBRT) on thrombin generation and platelet aggregation. We included 110 patients with localized NSCLC treated with SBRT. Blood samples were obtained prior to SBRT, immediately after SBRT completion, and 4-6 weeks following SBRT. Ex vivo and in vivo thrombin generations were analyzed using a calibrated automated thrombogram and commercial enzyme-linked immunosorbent assays. Platelet aggregation was evaluated using multiple electrode aggregometry. No significant differences were found in ex vivo or in vivo thrombin generation between blood samples before and immediately after SBRT treatment. Platelet aggregation was lower immediately after SBRT than before SBRT (TRAP: P = 0.04 and ASPI: P = 0.02) but remained within the reference interval. SBRT did not affect in vivo and ex vivo thrombin generation or platelet aggregation. SBRT did not cause prothrombotic changes in the coagulation in this study population of SBRT-treated patients with localized NSCLC.

OBJECTIVE To describe the relationship, through a scoping review, between thrombin generation assay (TGA) parameters and venous thromboembolism (VTE). MATERIAL AND METHODS A systematic search was carried out in the PubMed/MEDLINE, Embase, Cochrane, Virtual Health Library, Web of Science, and Scopus databases using descriptors for VTE and thrombin generation. Human studies assessing the association between TGA parameters and the occurrence of VTE were included. Screening of the title/abstract and full text was carried out independently by two researchers and divergent classifications were resolved by a third examiner. RESULTS 5204 studies were found, of which 90 were included after screening and full reading. The articles were grouped according to the study population and the factors associated with VTE: unprovoked VTE [33], cancer [20], hereditary/acquired thrombophilias [12], trauma/hospitalization [11], use of hormone replacement therapy/combined oral contraceptives [6], pregnancy [4] and use of anticoagulants [4]. The studies included were predominantly conducted in European countries between 2001 and 2024. Most of the studies were prospective cohort studies and used the Calibrated Automated Thrombogram (CAT) method to perform the TGA. The TGA parameters evaluated were mainly ETP and peak, which were elevated in patients with VTE in all the subgroups assessed. The TGA was able to predict the risk of primary and recurrent VTE and proved to be a useful tool for managing therapeutic regimens with anticoagulants in patients who suffered VTE. CONCLUSION The included studies demonstrated that TGA is a promising test in the context of VTE.

… MM treatment is also associated with increased thrombotic risk … Thus, thrombotic events in MM patients are multifactorial, … – thromboelastography (TEG), thrombin generation test (TGT) …

Introduction The Padua prediction score is a risk assessment model used to identify medical patients at high risk for venous thromboembolim (VTE).We aimed to assess the relationship …

… We sought to establish such data by looking for a correlation between thrombin generation … prostate cancer. We recently documented an increased rate of thrombin generation in such …

… , cancer, antithrombin congenital deficiency or the antiphospholipid antibody syndrome. Although only capillary blood for D-dimer testing was required, thrombosis … vein thrombosis was …

… thrombin formation, triggered by ruptured or eroded coronary atheroma. We investigated whether thrombin generation … (TF) initiated thrombin generation was assessed both computation…