Abstract 522: Activated FXI Regulates the Catalytic Activity of Adamts13 by Removing the CUB Domains

2017 ◽  
Vol 37 (suppl_1) ◽  
Author(s):  
Cristina Puy ◽  
Kathleen S Garland ◽  
Toshiaki Shirai ◽  
Stephanie E. Reitsma ◽  
Jevgenia Zilberman-Rudenko ◽  
...  
Keyword(s):  
Thrombus Formation ◽  
Adamts13 Activity ◽  
Binding Interaction ◽  
Vein Thrombosis ◽  
Cub Domain ◽  
A Cell ◽  
Von Willebrand ◽  
Deep Vein ◽  
Willebrand Factor

Background: ADAMTS13 cleaves and inactivates von Willebrand factor (VWF), which binds collagen, facilitating platelet adhesion under vascular injury. But is still uncertain how ADAMTS13 activity is regulated. Thrombin and plasmin have been shown to cleave ADAMTS13. Based on the fact that elevated levels of FXI is an independent risk factor for deep vein thrombosis and ischemic stroke, we hypothesize that FXIa inactivates ADAMTS13 leading to platelet aggregation and thrombus formation. Aim: To determine the functional role of inactivation of ADAMTS13 by FXIa. Methods and Results: Recombinant ADAMTS13 (250 nM) was incubated with FXIa (50 nM) for increasing times (0-3 hours) at 37 o C before being analyzed by western blot using an anti-ADAMTS13 antibody against the two CUB domains (C-terminal) or against the metalloproteinase (MET) domain (N-terminal). Our results show that FXIa caused the disappearance of the ADAMTS13 band (~200 kDa) and the appearance of a band at ~150 kDa when the samples were analyzed with the anti-MET antibody and a ~50 kDa band when the samples were analyzed with the anti-CUB antibody. The presence of aprotinin, which inhibits FXIa activity, blocked the degradation of ADAMTS13. Kallikrein or FXIIa were unable to cleave ADAMTS13. Using a cell surface immunoassay we observed that after incubation with FXIa, the detection of the CUB domain from ADAMTS13 was lost from endothelial cells surface. The incubation of ADAMTS13 with FXIa caused an increase in ADAMTS13 activity as measured by a fluorogenic substrate (FRETS). Conclusion: ADAMTS13 circulates in a closed conformation, which is maintained by a CUB-spacer domain binding interaction. ADAMTS13 becomes conformationally activated through interaction of its CUBs domains with VWF. Here we show that FXIa-mediated deletion of ADAMTS13-CUB domains enhances its capacity to cleave FRETS and blocks the interaction with VWF. Our results suggest that FXIa may limit ADAMTS13-mediated VWF inactivation.

Role of clotting factor VIII in effect of von Willebrand factor on occurrence of deep-vein thrombosis

The Lancet ◽  
1995 ◽  
Vol 345 (8943) ◽  
pp. 152-155 ◽  
Author(s):  
T. Koster ◽  
J.P. Vandenbroucke ◽  
F.R. Rosendaal ◽  
E. Briët ◽  
F.R. Rosendaal ◽  
...  
Keyword(s):  
Deep Vein Thrombosis ◽  
Factor Viii ◽  
Von Willebrand Factor ◽  
Clotting Factor ◽  
Vein Thrombosis ◽  
Von Willebrand ◽  
Deep Vein ◽  
Willebrand Factor

scholarly journals von Willebrand Factor Is a Critical Mediator of Deep Vein Thrombosis in a Mouse Model of Diet-Induced Obesity

2020 ◽  
Vol 40 (12) ◽  
pp. 2860-2874 ◽  
Author(s):  
Alison Michels ◽  
Courtney N. Dwyer ◽  
Jeffrey Mewburn ◽  
Kate Nesbitt ◽  
Charlotte Kawecki ◽  
...  
Keyword(s):  
Inferior Vena ◽  
Vena Cava ◽  
Obese Mice ◽  
Vein Thrombosis ◽  
Diet Induced Obesity ◽  
Von Willebrand ◽  
Deep Vein ◽  
Willebrand Factor ◽  
Vena Cava Stenosis

Objective: Obesity is characterized by chronic low-grade inflammation and consequentially a hypercoagulable state, associating with an increased incidence of venous thromboembolism. Increased VWF (von Willebrand factor) plasma concentration and procoagulant function are independent risk factors for venous thromboembolism and are elevated in obese patients. Here, we explore the pathobiological role of VWF in obesity-associated venous thrombosis using murine models. Approach and Results: We first showed that diet-induced obese mice have increased VWF plasma levels and FVIII (factor VIII) activity compared with littermate controls. Elevated VWF levels appeared to be because of both increased synthesis and impaired clearance. Diet-induced obesity-associated venous thrombosis was assessed using the inferior vena cava-stenosis model of deep vein thrombosis. Diet-induced obese mice developed larger venous thrombi that were rich in VWF, erythrocytes, and leukocytes. Administering a polyclonal anti-VWF antibody or an anti-VWF A1 domain nanobody was protective against obesity-mediated thrombogenicity. Delayed administration (3 hours post–inferior vena cava stenosis) similarly reduced thrombus weight in diet-induced obese mice. Conclusions: This study demonstrates the critical role of VWF in the complex, thrombo-inflammatory state of obesity. It adds to the growing rationale for targeting VWF-specific interactions in thrombotic disease.

scholarly journals Soluble P-Selectin, Von Willebrand Factor, And Adamts13 Levels As Risk Factors Of Deep Vein Thrombosis In Cancer Patients Undergoing Chemotherapy

2020 ◽  
Author(s):  
Budi Setiawan ◽  
Cecilia Oktaria Permatadewi ◽  
Baringin de Samakto ◽  
Ashar Bugis ◽  
Eko Adhi Pangarsa ◽  
...  
Keyword(s):  
Risk Factors ◽  
Deep Vein Thrombosis ◽  
Cancer Patients ◽  
Plasma Levels ◽  
Vein Thrombosis ◽  
Soluble P ◽  
Von Willebrand ◽  
Deep Vein ◽  
Willebrand Factor

Abstract Background There is a high number of deep vein thrombosis (DVT) incidence among cancer patients undergoing chemotherapy. Chemotherapy-induced vascular endothelial cell activation (VECA) is marked with increasing plasma levels of von Willebrand Factor (VWF) and soluble P-selectin (sP-selectin) leading to activation of endothelial cells and coagulation cascade. The biological role of a disintegrin-like and metalloproteinase with thrombospondin type 1, motif 13 (ADAMTS13) is to control the activity of VWF. The objectives of this study is to investigate the role of sP-selectin, VWF, and ADAMTS13 as risk factors for the incidence of DVT in cancer patients undergoing chemotherapy. Methods This prospective cohort study was conducted in Dr. Kariadi hospital, Semarang Indonesia, on 40 cancer patients. Soluble P-selectin, VWF, and ADAMS13 plasma levels were determined with enzyme-linked immunosorbent assay (ELISA) method, examined before and after chemotherapy. These patients were observed for the possibility of developing DVT during three months. Results Deep vein thrombosis was confirmed in 5 patients (12.5%) after a median period of 8 weeks. In patients with DVT, sP-selectin and VWF were significantly higher, while ADAMTS13 were significantly lower compared in cancer patients without DVT. Pre- and post-chemotherapy concentration of sP selectin, VWF, and ADAMTS13 could effectively predict the incidence of DVT in cancer patients undergoing chemotherapy. The levels of sP-selectin, VWF and ADAMTS13 pre-chemotherapy with cut-off point >106.7 ng/mL, >2.99 U/mL and <0,80 U/mL, respectively, had relative risk (RR) for DVT incidence being 16 (95% CI 2,06-124,25, p=0,001); 36 (95% CI 5,21-248,65, p=0,000) and 10,5 (95% CI 1,31-84,28, p=0,015), respectively, whereas the levels of sP-selectin, VWF and ADAMTS13 post-chemotherapy with cut-off point >111.7 ng/mL, >3,06 U/mL and <0,49 U/mL, respectively, had RR for DVT incidence being 8.7 (95% CI 1,01-74,39, p=0,045); 20,4 (95% CI 2,60-159,94, p=0,004) and 26,25 (95% CI 3,50-196,48, p=0,002), respectively. Pre-chemotherapy vWF levels (cut-off value >2.99 U/mL) was found to be independently predict DVT incidence with RR 11.1 (95% CI, 1.95-62.74, p=0.007). Conclusions Plasma levels of VWF more than 2.99 U/mL pre-chemotherapy was an independent risk factor for DVT incidence, which could be performed early and helpful for thromboprophylaxis therapy.

The role of platelet von Willebrand factor in platelet adhesion and thrombus formation: a study of 34 patients with various subtypes of type I von Willebrand disease

1994 ◽  
Vol 86 (2) ◽  
pp. 327-332 ◽  
Author(s):  
Edith Fressinaud ◽  
Augusto B. Federici ◽  
Giancarlo Castaman ◽  
Chantal Rothschild ◽  
Francesco Rodeghiero ◽  
...  
Keyword(s):  
Von Willebrand Factor ◽  
Platelet Adhesion ◽  
Thrombus Formation ◽  
Von Willebrand Disease ◽  
Type I ◽  
Von Willebrand ◽  
Willebrand Factor

scholarly journals The distal carboxyl-terminal domains of ADAMTS13 are required for regulation of in vivo thrombus formation

Blood ◽  
2009 ◽  
Vol 113 (21) ◽  
pp. 5323-5329 ◽  
Author(s):  
Fumiaki Banno ◽  
Anil K. Chauhan ◽  
Koichi Kokame ◽  
Jin Yang ◽  
Shigeki Miyata ◽  
...  
Keyword(s):  
Shear Rate ◽  
Genetic Background ◽  
Thrombus Formation ◽  
Short Form ◽  
Von Willebrand ◽  
Willebrand Factor ◽  
Terminal Domains

Abstract ADAMTS13 is a multidomain protease that limits platelet thrombogenesis through the cleavage of von Willebrand factor (VWF). We previously identified 2 types of mouse Adamts13 gene: the 129/Sv-strain Adamts13 gene encodes the long-form ADAMTS13 having the same domains as human ADAMTS13, whereas the C57BL/6-strain Adamts13 gene encodes the short-form ADAMTS13 lacking the distal C-terminal domains. To assess the physiologic significance of the distal C-terminal domains of ADAMTS13, we generated and analyzed 129/Sv-genetic background congenic mice (Adamts13S/S) that carry the short-form ADAMTS13. Similar to wild-type 129/Sv mice (Adamts13L/L), Adamts13S/S did not have ultralarge VWF multimers in plasma, in contrast to 129/Sv-genetic background ADAMTS13-deficient mice (Adamts13−/−). However, in vitro thrombogenesis under flow at a shear rate of 5000 s−1 was accelerated in Adamts13S/S compared with Adamts13L/L. Both in vivo thrombus formation in ferric chloride–injured arterioles and thrombocytopenia induced by collagen plus epinephrine challenge were more dramatic in Adamts13S/S than in Adamts13L/L but less than in Adamts13−/−. These results suggested that the C-terminally truncated ADAMTS13 exhibited decreased activity in the cleavage of VWF under high shear rate. Role of the C-terminal domains may become increasingly important under prothrombotic conditions.

Synergic effect of GPIBA and von Willebrand factor in pathogenesis of deep vein thrombosis

Vascular ◽  
2020 ◽  
Vol 28 (3) ◽  
pp. 309-313
Author(s):  
Da Li ◽  
Xiaosong Zhang ◽  
Honggang Zhang ◽  
Xiaoqiang Li
Keyword(s):  
Cardiovascular Disease ◽  
Deep Vein Thrombosis ◽  
Membrane Protein ◽  
Von Willebrand Factor ◽  
Integrin Subunit ◽  
Dynamic Changes ◽  
Vein Thrombosis ◽  
Von Willebrand ◽  
Deep Vein ◽  
Willebrand Factor

Objectives In cardiovascular disease, deep vein thrombosis is one of the vital symptoms causing pulmonary thromboembolism. However, the pathogenesis of deep vein thrombosis is still not clear. One of the critical factors leading to deep vein thrombosis is the platelet aggregation that is mediated by a set of key genes including platelet membrane protein coded by platelet glycoprotein Ib alpha chain (GPIBA). Methods Deep vein thrombosis model was established according to the previous protocol, and venous blood and thrombi were collected for further analysis. Results The dynamic changes of GPIBA and coagulation factor, von Willebrand factor, were observed in deep vein thrombosis models. Meanwhile, critical proteins participating in adhesion and binding of platelets such as epithelial membrane protein 2 (EMP2), vascular cell adhesion protein 1 (VCAM1), immunoreceptor tyrosine-based activation motif 1 (ITAM1), integrin subunit alpha M (ITGAM), or fibronectin were also differentially expressed in deep vein thrombosis models. Conclusions Application of heparin could reverse these dynamic changes in deep vein thrombosis models. Thus, we explained the potential synergic role of GPIBA and von Willebrand factor in regulating the occurrence of deep vein thrombosis and provide therapeutic target against cardiovascular disease.

Evaluation of ADAMTS13 Activity and Inflammatory Markers in Deep Venous Thrombosis Patients

Blood ◽  
2011 ◽  
Vol 118 (21) ◽  
pp. 5246-5246
Author(s):  
Bruna de Moraes Mazetto ◽  
Fernanda A. Orsi ◽  
Aline Barnabé ◽  
Erich V de Paula ◽  
Joyce M Annichino-Bizzacchi
Keyword(s):  
Inflammatory Markers ◽  
Conflicts Of Interest ◽  
Adamts13 Activity ◽  
Vein Thrombosis ◽  
Outpatient Unit ◽  
Inflammatory Conditions ◽  
Significant Difference ◽  
Von Willebrand ◽  
D Dimer

Abstract Abstract 5246 BACKGROUND: The role of inflammation on the pathophysiology of arterial thrombosis has been extensively studied. However, the relationship between inflammation and deep vein thrombosis (DVT) is not completely understood. Conflicting reports have been published about the levels of proteins involved in the inflammatory response in the context of DVT. Changes in ADAMTS13 levels have been reported in other inflammatory conditions such as sepsis, and this protease could be involved in the interplay between hemostasis and inflammation. OBJECTIVE: To evaluate ADAMTS13 activity in DVT patients and its association with inflammatory markers (IL-6, IL-8, CRP, TNF-α), D-dimer and von Willebrand Factor (VWF) levels. METHODOLOGY: Thirty-eight DVT patients, from 6 months to five years after the diagnosis of DVT, followed at the outpatient unit of thrombotic diseases from University of Campinas and 38 healthy volunteers selected as controls were included in the study. ADAMTS13 activity was determined by the residual binding of VWF to collagen. VWF, IL-6, IL-8 and TNF-alpha levels were determined by ELISA, and D-dimer levels was determined by a turbidimetric method. RESULTS: In this study population DVT was triggered by transient risk factors, particularly the use of oral contraceptives. No patient presented renal, hepatic or malignant disease. Median ADAMTS13 activity was not statistically different between patients (median: 98.2%; range: 70–146) and controls (median: 96.1%; range: 66–117; p=0.35). IL-6 and TNF-alfa levels were also higher in patients (median: 1.0pg/mL and 2.3pg/mL) compared to controls (median: 0.64pg/mL and 1.7pg/mL, p=0,01 and p=0,009). In addition, VWF and D-dimer levels were also significantly higher in patients (all P<0.01). No significant difference could be demonstrated between IL-8 and CRP levels from patients and controls. No significant correlation between ADAMTS13 levels and other inflammatory markers could be demonstrated. VWF levels correlated significantly with IL-8 (r=-0.4635, p< 0.0001). CONCLUSIONS: This study reinforces the role of inflammation in the pathogenesis of DVT. The correlation of VWF and IL-8 supports the notion that IL-8 stimulates the secretion of VWF. The role of ADAMTS13 in the context of DVT is yet to be determined. Disclosures: No relevant conflicts of interest to declare.

Altered thrombus formation in von Willebrand factor–deficient mice expressing von Willebrand factor variants with defective binding to collagen or GPIIbIIIa

Blood ◽  
2008 ◽  
Vol 112 (3) ◽  
pp. 603-609 ◽  
Author(s):  
Isabelle Marx ◽  
Olivier D. Christophe ◽  
Peter J. Lenting ◽  
Alain Rupin ◽  
Marie-Odile Vallez ◽  
...  
Keyword(s):  
Von Willebrand Factor ◽  
Thrombus Formation ◽  
Vessel Occlusion ◽  
Injury Model ◽  
Von Willebrand ◽  
Willebrand Factor ◽  
Deficient Mice ◽  
Interesting Alternative ◽  
Model Expression

Abstract The role of von Willebrand factor (VWF) in thrombosis involves its binding to a number of ligands. To investigate the relative importance of these particular interactions in the thrombosis process, we have introduced mutations into murine VWF (mVWF) cDNA inhibiting VWF binding to glycoprotein (Gp) Ib, GPIIbIIIa, or to fibrillar collagen. These VWF mutants were expressed in VWF-deficient mice (VWF−/−) by using an hydrodynamic injection approach, and the mice were studied in the ferric chloride–induced injury model. Expression of the collagen and the GPIIbIIIa VWF-binding mutants in VWF−/− mice resulted in delayed thrombus growth and significantly increased vessel occlusion times compared with mice expressing wild-type (WT) mVWF (30 ± 3 minutes and 38 ± 4 minutes for the collagen and GPIIbIIIa mutants, respectively, vs 19 ± 3 minutes for WT mVWF). Interestingly, these mutants were able to correct bleeding time as efficiently as WT mVWF. In contrast, VWF−/− mice expressing the GPIb binding mutant failed to restore thrombus formation and were bleeding for as long as they were observed, confirming the critical importance of the VWF-GPIb interaction. Our observations suggest that targeting the VWF-collagen or VWF-GPIIbIIIa interactions could be an interesting alternative for new antithrombotic strategies.

scholarly journals Synergism between platelet collagen receptors defined using receptor-specific collagen-mimetic peptide substrata in flowing blood

Blood ◽  
2010 ◽  
Vol 115 (24) ◽  
pp. 5069-5079 ◽  
Author(s):  
Nicholas Pugh ◽  
Anna M. C. Simpson ◽  
Peter A. Smethurst ◽  
Philip G. de Groot ◽  
Nicolas Raynal ◽  
...  
Keyword(s):  
Thrombus Formation ◽  
Confocal Imaging ◽  
Peptide Substrates ◽  
Shear Rates ◽  
Glycoprotein Vi ◽  
Collagen Receptors ◽  
Platelet Binding ◽  
Von Willebrand ◽  
Willebrand Factor

AbstractExposed subendothelial collagen acts as a substrate for platelet adhesion and thrombus formation after vascular injury. Synthetic collagen-derived triple-helical peptides, designated collagen-related peptide (CRP), GFOGER, and VWF-III, can specifically engage the platelet collagen receptors, glycoprotein VI and integrin α2β1, and plasma von Willebrand factor (VWF), respectively. Hitherto, the role of these 3 collagen-binding axes has been studied indirectly. Use of these uniform peptide substrates, rather than collagen fibers, provides independent control of each axis. Here, we use confocal imaging and novel image analysis techniques to investigate the effects of receptor-ligand engagement on platelet binding and activation during thrombus formation under flow conditions. At low shear (100s−1 and 300s−1), both GFOGER and CRP are required for thrombus formation. At 1000s−1, a combination of either CRP or GFOGER with VWF-III induces comparable thrombus formation, and VWF-III increases thrombus deposition at all shear rates, being indispensable at 3000s−1. A combination of CRP and VWF-III is sufficient to support extensive platelet deposition at 3000s−1, with slight additional effect of GFOGER. Measurement of thrombus height after specific receptor blockade or use of altered proportions of peptides indicates a signaling rather than adhesive role for glycoprotein VI, and primarily adhesive roles for both α2β1 and the VWF axis.

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