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Evsen A, Aktan A, Kılıç R, Özbek M. Association Between ABO Blood Group, Peripheral Artery Disease Lesion Severity, and Coronary Artery Disease Coexistence. Ann Vasc Surg 2025; 113:74-82. [PMID: 39855381 DOI: 10.1016/j.avsg.2024.12.067] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/17/2024] [Revised: 12/17/2024] [Accepted: 12/23/2024] [Indexed: 01/27/2025]
Abstract
BACKGROUND This study aims to investigate the relationship between ABO blood groups and the severity of peripheral artery disease (PAD) lesions, the coexistence of coronary artery disease (CAD) with PAD, and to identify which blood groups may be more predisposed to these conditions. METHODS This study, which has a single-center and retrospective design, includes 305 patients diagnosed with peripheral artery disease (PAD) between 2015 and 2021. The patients were selected from those with at least 50% stenosis detected by computed tomography (CT) angiography. The severity of PAD lesions was evaluated according to the TransAtlantic Inter-Society Consensus (TASC-II) classification, and the presence of CAD was determined by simultaneous conventional coronary angiography. Demographic data, comorbidities, and laboratory results were collected, and statistical analyses were performed using chi-square tests, logistic regression, and t-tests. All of these patients were divided into 2 groups based on their ABO blood types: O and non-O blood groups. RESULTS This study demonstrates a significant relationship between blood group classification and both the severity of PAD and the prevalence of CAD. Patients with non-O blood groups exhibited a higher likelihood of having severe PAD lesions (TASC-C and TASC-D) compared to those with O blood groups, who more frequently had milder lesions (TASC-A and TASC-B) (P < 0.001). The prevalence of CAD was also significantly higher among patients with non-O blood groups than those with the O blood group (54.4% vs. 36.4%; P = 0.003). In the logistic regression analysis, non-O blood groups, alongside age, hypertension (HT), and chronic kidney disease (CKD), emerged as independent predictors of severe PAD (P < 0.001 for non-O blood groups). Similarly, older age and non-O blood group status were identified as significant independent predictors of CAD (P = 0.004). These findings suggest that blood group classification, particularly non-O groups, may influence the severity and coexistence of PAD and CAD. CONCLUSION Our study reveals a significant association between ABO blood groups and both the severity of PAD and the association of CAD, and shows that non-O blood groups are linked to more severe forms of these conditions. These findings highlight the potential role of ABO blood groups in cardiovascular risk stratification, suggesting a need for further research to confirm these associations and understand their clinical implications.
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Affiliation(s)
- Ali Evsen
- Dağkapı State Hospital, Department of Cardiology, Diyarbakır, Turkey.
| | - Adem Aktan
- Mardin Artuklu University Faculty of Medicine, Department of Cardiology, Mardin, Turkey
| | - Raif Kılıç
- Mardin Artuklu University Faculty of Medicine, Department of Cardiology, Mardin, Turkey
| | - Mehmet Özbek
- Department of Cardiology, Dicle University School of Medicine, Diyarbakır, Turkey
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Interlandi G. Exploring ligands that target von Willebrand factor selectively under oxidizing conditions through docking and molecular dynamics simulations. Proteins 2024; 92:1261-1275. [PMID: 38829206 PMCID: PMC11471382 DOI: 10.1002/prot.26706] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/29/2024] [Revised: 04/25/2024] [Accepted: 05/13/2024] [Indexed: 06/05/2024]
Abstract
The blood protein von Willebrand factor (VWF) is a large multimeric protein that, when activated, binds to blood platelets, tethering them to the site of vascular injury and initiating blood coagulation. This process is critical for the normal hemostatic response, but especially under inflammatory conditions, it is thought to be a major player in pathological thrombus formation. For this reason, VWF has been the target for the development of anti-thrombotic therapeutics. However, it is challenging to prevent pathological thrombus formation while still allowing normal physiological blood coagulation, as currently available anti-thrombotic therapeutics are known to cause unwanted bleeding, in particular intracranial hemorrhage. This work explores the possibility of inhibiting VWF selectively under the inflammatory conditions present during pathological thrombus formation. In particular, the A2 domain of VWF is known to inhibit the neighboring A1 domain from binding to the platelet surface receptor GpIbα, and this auto-inhibitory mechanism has been shown to be removed by oxidizing agents released during inflammation. Hence, finding drug molecules that bind at the interface between A1 and A2 only under oxidizing conditions could restore such an auto-inhibitory mechanism. Here, by using a combination of computational docking, molecular dynamics simulations, and free energy perturbation calculations, a ligand from the ZINC15 database was identified that binds at the A1A2 interface, with the interaction being stronger under oxidizing conditions. The results provide a framework for the discovery of drug molecules that bind to a protein selectively in the presence of inflammatory conditions.
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Affiliation(s)
- Gianluca Interlandi
- Department of Bioengineering, University of Washington, Seattle, Washington, USA
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3
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Sarani N, Dasgupta A, Enders M, Rowan L, Elsarraj H, Gralnek S, Shay M, Lemar LR, Simpson SQ, Cunningham MT, Zheng XL. Clinical Utility of Recently Food and Drug Administration-Approved IntelliSep Test (Sepsis Biomarker) for Early Diagnosis of Sepsis: Comparison with Other Biomarkers. J Clin Med 2024; 13:4852. [PMID: 39200994 PMCID: PMC11355800 DOI: 10.3390/jcm13164852] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/29/2024] [Revised: 08/13/2024] [Accepted: 08/14/2024] [Indexed: 09/02/2024] Open
Abstract
Context: IntelliSep by Cytovale has received United States (U.S.) Food and Drug Administration (FDA) approval as a sepsis biomarker test. However, the clinical utility of this new test is not assessed in emergency departments. Objective: We investigated the clinical utility of this test using 44 patients visiting the emergency department at The University of Kansas Medical Center by comparing it with the monocyte distribution width (MDW) and other biomarkers including the von Willebrand factor (vWF) and ADAMTS13. Design and Methods: IntelliSep assesses the cellular host response via deformability cytometry of biophysical leukocyte properties and produces a score (IntelliSep Index; ISI: from 0.1 (lowest risk) to 10 (highest risk). We measured the ISI in 44 patients (19 high probability and 25 low probability of sepsis groups) using EDTA-anticoagulated blood. Left over plasma was used for measuring the plasma von Willebrand factor (vWF) and ADAMTS13 antigen by ELISA assays. The MDW was obtained during routine CBC analysis using a Beckman hematology analyzer. The lactate and high-sensitivity troponin I levels were measured using a Beckman analyzer. Procalcitonin was measured using a Cobas e801 analyzer. Results: The median ISI was twofold higher in the high-probability group than in the low-probability group (p < 0.01) while the median MDW was 34.5% higher in the high-probability group than in the low-probability group (p < 0.01). However, the correlation between the ISI and MDW was only modest (r = 0.66). In addition, significantly higher levels of plasma vWF antigen but lower levels of plasma ADAMTS13 antigen in the high-probability group were found, resulting in significantly higher vWF/ADAMTS13 ratios in the high-probability group than in the low-probability group. Conclusions: The new IntelliSep test along with vWF/ADAMTS13 ratios may be useful for the early diagnosis of sepsis in patients visiting the emergency department, which appears to be superior to the traditional marker, MDW.
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Affiliation(s)
- Nima Sarani
- Department of Emergency Medicine, The University of Kansas Medical Center, Kansas City, KS 66160, USA; (N.S.); (M.E.); (L.R.); (L.R.L.)
| | - Amitava Dasgupta
- Department of Pathology and Laboratory Medicine, The University of Kansas Medical Center, Kansas City, KS 66160, USA; (A.D.); (H.E.); (S.G.); (M.S.); (M.T.C.)
| | - Maria Enders
- Department of Emergency Medicine, The University of Kansas Medical Center, Kansas City, KS 66160, USA; (N.S.); (M.E.); (L.R.); (L.R.L.)
| | - Lauren Rowan
- Department of Emergency Medicine, The University of Kansas Medical Center, Kansas City, KS 66160, USA; (N.S.); (M.E.); (L.R.); (L.R.L.)
| | - Hanan Elsarraj
- Department of Pathology and Laboratory Medicine, The University of Kansas Medical Center, Kansas City, KS 66160, USA; (A.D.); (H.E.); (S.G.); (M.S.); (M.T.C.)
| | - Sarah Gralnek
- Department of Pathology and Laboratory Medicine, The University of Kansas Medical Center, Kansas City, KS 66160, USA; (A.D.); (H.E.); (S.G.); (M.S.); (M.T.C.)
| | - Madison Shay
- Department of Pathology and Laboratory Medicine, The University of Kansas Medical Center, Kansas City, KS 66160, USA; (A.D.); (H.E.); (S.G.); (M.S.); (M.T.C.)
| | - Lucas R. Lemar
- Department of Emergency Medicine, The University of Kansas Medical Center, Kansas City, KS 66160, USA; (N.S.); (M.E.); (L.R.); (L.R.L.)
| | - Steven Q. Simpson
- Department of Internal Medicine, The University of Kansas Medical Center, Kansas City, KS 66160, USA;
| | - Mark T. Cunningham
- Department of Pathology and Laboratory Medicine, The University of Kansas Medical Center, Kansas City, KS 66160, USA; (A.D.); (H.E.); (S.G.); (M.S.); (M.T.C.)
| | - X. Long Zheng
- Department of Pathology and Laboratory Medicine, The University of Kansas Medical Center, Kansas City, KS 66160, USA; (A.D.); (H.E.); (S.G.); (M.S.); (M.T.C.)
- Institute of Reproductive Medicine and Developmental Sciences, The University of Kansas Medical Center, Kansas City, KS 66160, USA
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Interlandi G. Exploring ligands that target von Willebrand factor selectively under oxidizing conditions through docking and molecular dynamics simulations. BIORXIV : THE PREPRINT SERVER FOR BIOLOGY 2024:2024.03.22.586354. [PMID: 38585752 PMCID: PMC10996496 DOI: 10.1101/2024.03.22.586354] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Grants] [Track Full Text] [Subscribe] [Scholar Register] [Indexed: 04/09/2024]
Abstract
The blood protein von Willebrand factor (VWF) is a large multimeric protein that, when activated, binds to blood platelets tethering them to the site of vascular injury initiating blood coagulation. This process is critical for the normal haemostatic response, but especially under inflammatory conditions it is thought to be a major player in pathological thrombus formation. For this reason, VWF has been the target for the development of anti-thrombotic therapeutics. However, it is challenging to prevent pathological thrombus formation while still allowing normal physiological blood coagulation as currently available anti-thrombotic therapeutics are known to cause unwanted bleeding in particular intracranial haemorrhage. This work explores the possibility of inhibiting VWF selectively under the inflammatory conditions present during pathological thrombus formation. In particular, the A2 domain of VWF is known to inhibit the neighboring A1 domain from binding to the platelet surface receptor GpIbα and this auto-inhibitory mechanism has been shown to be removed by oxidizing agents released during inflammation. Hence, finding drug molecules that bind at the interface between A1 and A2 only under oxidizing conditions could restore such auto-inhibitory mechanism. Here, by using a combination of computational docking, molecular dynamics simulations and free energy perturbation calculations, a ligand from the ZINC15 database was identified that binds at the A1A2 interface with the interaction being stronger under oxidizing conditions. The results provide a framework for the discovery of drug molecules that bind to a protein selectively in inflammatory conditions.
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Álvarez Román MT, Rivas Pollmar MI, De la Corte-Rodríguez H, Gómez-Cardero P, Rodríguez-Merchán EC, Gutiérrez-Alvariño M, García-Pérez E, Martín-Salces M, Zagrean D, Butta-Coll NV, Jiménez-Yuste V. Knee replacement surgery in a patient with acquired von Willebrand disease: a case study with recommendations for patient management. Ann Med Surg (Lond) 2024; 86:1681-1686. [PMID: 38463081 PMCID: PMC10923270 DOI: 10.1097/ms9.0000000000001690] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/01/2023] [Accepted: 12/27/2023] [Indexed: 03/12/2024] Open
Abstract
Introduction and importance Acquired von Willebrand disease (AvWD) is a rare underdiagnosed bleeding disorder caused by alterations in the levels of the major blood-clotting protein von Willebrand factor (vWF). The clinical and laboratory parameters of AvWD are similar to congenital vWD, but it is found in individuals with no positive family history with no underlying genetic basis. The disease remains multifactorial and incompletely understood. Proposed mechanisms include the development of autoantibodies to vWF, absorption of high molecular weight vWF multimers that impair normal function, shear stress induced vWF cleavage and increased proteolysis.The aetiology of the disease is variable, the most common being hematoproliferation, lymophoproliferation, myeloproliferation and autoimmune and cardiovascular disorders. Consensus and protocols for AvWD patients that require major surgery are currently lacking. Patients with AvWD can experience thrombotic events during surgery as a result of therapeutic interactions with pro-thrombotic risk factors. Case presentation Here, the authors report a patient with AvWD requiring a knee prosthesis implantation due to chronic pain, limited range of motion and functional impairment. The patient had a high risk of bleeding during surgery and was at risk of thrombosis due to age and obesity. Clinical discussion Perioperative care required a collaborative approach and the management of bleeding. The patient was administered vWF concentrate Willfact lacking Factor VIII to prevent haemorrhage and to minimize the risk of thrombosis. Conclusion The treatment was effective and well-tolerated. The authors use this information to provide recommendations for AvWD patients for whom major surgery is indicated.
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Affiliation(s)
| | | | | | | | - E. Carlos Rodríguez-Merchán
- Osteoarticular Surgery Research, Hospital La Paz Institute for Health Research, IdiPAZ (La Paz University Hospital—Autonomous University of Madrid)
| | | | | | | | | | | | - Víctor Jiménez-Yuste
- Departments ofHematology
- Department of Medicine, Autonomous University of Madrid, Madrid, Spain
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Ricco C, Eldaboush A, Liu ML, Werth VP. Extracellular Vesicles in the Pathogenesis, Clinical Characterization, and Management of Dermatomyositis: A Narrative Review. Int J Mol Sci 2024; 25:1967. [PMID: 38396646 PMCID: PMC10889219 DOI: 10.3390/ijms25041967] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/13/2023] [Revised: 02/01/2024] [Accepted: 02/02/2024] [Indexed: 02/25/2024] Open
Abstract
Extracellular vesicles (EVs) are lipid-bilayer particles secreted from cells that primarily assist in cell-to-cell communication through the content of their cargo, such as proteins and RNA. EVs have been implicated in the pathogenesis of various autoimmune diseases, including dermatomyositis (DM), an inflammatory autoimmune disease characterized by distinct cutaneous manifestations, myopathy, and lung disease. We sought to review the role of EVs in DM and understand how they contribute to the pathogenesis and clinical characterization of the disease. We summarized the research progress on EVs in dermatomyositis based on recent publications. EV cargoes, such as double-stranded DNA, microRNA, and proteins, contribute to DM pathogenesis and mediate the proinflammatory response and cytokine release through signaling pathways such as the stimulator of interferon genes (STING) pathway. These nucleic acids and proteins have been proposed as disease-specific, stable biomarkers to monitor disease activity and responses to therapy. They also correlate with clinical parameters, inflammatory markers, and disease severity scores. Furthermore, some markers show an association with morbidities of DM, such as muscle weakness and interstitial lung disease. The continued study of EVs will help us to further elucidate our understanding of dermatomyositis.
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Affiliation(s)
- Cristina Ricco
- Corporal Michael J. Crescenz Veterans Affairs Medical Center, Philadelphia, PA 19104, USA; (C.R.); (A.E.); (M.-L.L.)
- Department of Dermatology, School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA
| | - Ahmed Eldaboush
- Corporal Michael J. Crescenz Veterans Affairs Medical Center, Philadelphia, PA 19104, USA; (C.R.); (A.E.); (M.-L.L.)
- Department of Dermatology, School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA
| | - Ming-Lin Liu
- Corporal Michael J. Crescenz Veterans Affairs Medical Center, Philadelphia, PA 19104, USA; (C.R.); (A.E.); (M.-L.L.)
- Department of Dermatology, School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA
| | - Victoria P. Werth
- Corporal Michael J. Crescenz Veterans Affairs Medical Center, Philadelphia, PA 19104, USA; (C.R.); (A.E.); (M.-L.L.)
- Department of Dermatology, School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA
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Alfano DN, Miller MJ, Bubeck Wardenburg J. Endothelial ADAM10 utilization defines a molecular pathway of vascular injury in mice with bacterial sepsis. J Clin Invest 2023; 133:e168450. [PMID: 37788087 PMCID: PMC10688991 DOI: 10.1172/jci168450] [Citation(s) in RCA: 3] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/04/2023] [Accepted: 09/28/2023] [Indexed: 10/05/2023] Open
Abstract
The endothelium plays a critical role in the host response to infection and has been a focus of investigation in sepsis. While it is appreciated that intravascular thrombus formation, severe inflammation, and loss of endothelial integrity impair tissue oxygenation during sepsis, the precise molecular mechanisms that lead to endothelial injury remain poorly understood. We demonstrate here that endothelial ADAM10 was essential for the pathogenesis of Staphylococcus aureus sepsis, contributing to α-toxin-mediated (Hla-mediated) microvascular thrombus formation and lethality. As ADAM10 is essential for endothelial development and homeostasis, we examined whether other major human sepsis pathogens also rely on ADAM10-dependent pathways in pathogenesis. Mice harboring an endothelium-specific knockout of ADAM10 were protected against lethal Pseudomonas aeruginosa and Streptococcus pneumoniae sepsis, yet remained fully susceptible to group B streptococci and Candida albicans sepsis. These studies illustrate a previously unknown role for ADAM10 in sepsis-associated endothelial injury and suggest that understanding pathogen-specific divergent host pathways in sepsis may enable more precise targeting of disease.
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Affiliation(s)
| | - Mark J. Miller
- Division of Infectious Diseases, Department of Internal Medicine, Washington University School of Medicine, St. Louis, Missouri, USA
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Belyaev AV, Kushchenko YK. Biomechanical activation of blood platelets via adhesion to von Willebrand factor studied with mesoscopic simulations. Biomech Model Mechanobiol 2023; 22:785-808. [PMID: 36627458 PMCID: PMC9838538 DOI: 10.1007/s10237-022-01681-3] [Citation(s) in RCA: 9] [Impact Index Per Article: 4.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/22/2022] [Accepted: 12/22/2022] [Indexed: 01/12/2023]
Abstract
Platelet adhesion and activation are essential initial processes of arterial and microvascular hemostasis, where high hydrodynamic forces from the bloodflow impede coagulation. The process relies on von Willebrand factor (VWF)-a linear multimeric protein of blood plasma plays a pivotal role in mechanochemical regulation of shear-induced platelet aggregation (SIPA). Adhesive interactions between VWF and glycoprotein receptors GPIb are crucial for platelet recruitment under high shear stress in fluid. Recent advances in experimental studies revealed that mechanical tension on the extracellular part of GPIb may trigger a cascade of biochemical reactions in platelets leading to activation of integrins [Formula: see text] (also known as GPIIb/IIIa) and strengthening of the adhesion. The present paper is aimed at investigation of this process by three-dimensional computer simulations of platelet adhesion to surface-grafted VWF multimers in pressure-driven flow of platelet-rich plasma. The simulations demonstrate that GPIb-mediated mechanotransduction is a feasible way of platelet activation and stabilization of platelet aggregates under high shear stress. Quantitative understanding of mechanochemical processes involved in SIPA would potentially promote the discovery of new anti-platelet medication and the development of multiscale numerical models of platelet thrombosis and hemostasis.
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Affiliation(s)
- Aleksey V. Belyaev
- grid.14476.300000 0001 2342 9668Faculty of Physics, M.V. Lomonosov Moscow State University, 1-2 Leninskiye Gory, Moscow, Russia 119991
| | - Yulia K. Kushchenko
- grid.14476.300000 0001 2342 9668Faculty of Physics, M.V. Lomonosov Moscow State University, 1-2 Leninskiye Gory, Moscow, Russia 119991
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9
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van Paridon PCS, Panova‐Noeva M, van Oerle R, Schulz A, Prochaska JH, Arnold N, Schmidtmann I, Beutel M, Pfeiffer N, Münzel T, Lackner KJ, ten Cate H, Wild PS, Spronk HMH. Lower levels of vWF are associated with lower risk of cardiovascular disease. Res Pract Thromb Haemost 2022; 6:e12797. [PMID: 36381288 PMCID: PMC9637545 DOI: 10.1002/rth2.12797] [Citation(s) in RCA: 5] [Impact Index Per Article: 1.7] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/04/2022] [Revised: 07/23/2022] [Accepted: 07/29/2022] [Indexed: 11/09/2022] Open
Abstract
Objective The current study was undertaken to prospectively explore whether having low levels of von Willebrand factor (vWF) antigen and vWF activity reduce the risk for cardiovascular disease and death. Methods VWF antigen and vWF activity were measured by enzyme-linked immunosorbent assay and an immunological-based assay, respectively, in a subsample of 4857 individuals aged between 35 and 74 years old, enrolled between April 2007 and October 2008 in the population-based Gutenberg Health Study. VWF antigen and activity below the 20th percentile was set as a measure of "low vWF." Adjusted robust Poisson regression models were used to analyze the relation between low vWF and the incidence of cardiovascular disease (CVD). Consequent adjusted cox regression models as well as cumulative incidence plots were calculated to explore the relation between all-cause and cardiovascular mortality and low vWF. Results VWF activity levels <20th percentile (i.e., <76.2%) were associated with a decreased relative risk for CVD (RR: 0.59, 95% CI: 0.37-0.95), despite adjusting for age and sex. After adjusting for levels of F-VIII, the association persisted (RR: 0.60, 95% CI: 0.36-0.99). The cumulative incidence plots demonstrated that vWF antigen <20th percentile significantly correlated with decreased cardiovascular mortality. VWF antigen<20th percentile (i.e., <83%) was significantly associated with lower risk of all-cause mortality, despite adjusting for clinical factors (RR: 61, 95% CI: 0.41-0.91). Conclusion The study demonstrated that having low vWF activity levels were associated with a lower risk for CVD. Additionally, it revealed a decreased risk of cardiovascular and all-cause mortality in individuals with low levels of vWF antigen, shining new light on vWF as a potential target for novel therapies.
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Affiliation(s)
- Pauline C. S. van Paridon
- Laboratory for Clinical Thrombosis and Hemostasis, Department of Internal Medicine, Cardiovascular Research Institute Maastricht (CARIM)Maastricht University Medical CenterMaastrichtThe Netherlands
- Center for Thrombosis and Hemostasis (CTH)University Medical Center of the Johannes Gutenberg‐University MainzMainzGermany
| | - Marina Panova‐Noeva
- Center for Thrombosis and Hemostasis (CTH)University Medical Center of the Johannes Gutenberg‐University MainzMainzGermany
- DZHK (German Center for Cardiovascular Research)Partner Site RhineMainMainzGermany
| | - Rene van Oerle
- Laboratory for Clinical Thrombosis and Hemostasis, Department of Internal Medicine, Cardiovascular Research Institute Maastricht (CARIM)Maastricht University Medical CenterMaastrichtThe Netherlands
| | - Andreas Schulz
- Preventive Cardiology and Preventive Medicine, Center for CardiologyUniversity Medical Center of the Johannes Gutenberg‐University MainzMainzGermany
| | - Jürgen H. Prochaska
- Center for Thrombosis and Hemostasis (CTH)University Medical Center of the Johannes Gutenberg‐University MainzMainzGermany
- DZHK (German Center for Cardiovascular Research)Partner Site RhineMainMainzGermany
- Preventive Cardiology and Preventive Medicine, Center for CardiologyUniversity Medical Center of the Johannes Gutenberg‐University MainzMainzGermany
| | - Natalie Arnold
- Preventive Cardiology and Preventive Medicine, Center for CardiologyUniversity Medical Center of the Johannes Gutenberg‐University MainzMainzGermany
| | - Irene Schmidtmann
- Institute of Medical Biostatistics, Epidemiology and InformaticsUniversity Medical Center of the Johannes Gutenberg‐University MainzMainzGermany
| | - Manfred Beutel
- Department of Psychosomatic Medicine and PsychotherapyUniversity Medical Center of the Johannes Gutenberg‐University MainzMainzGermany
| | - Norbert Pfeiffer
- Department of OphthalmologyUniversity Medical Center of the Johannes Gutenberg‐University MainzMainzGermany
| | - Thomas Münzel
- DZHK (German Center for Cardiovascular Research)Partner Site RhineMainMainzGermany
- Center for Cardiology IUniversity Medical Center of the Johannes Gutenberg‐University MainzMainzGermany
| | - Karl J. Lackner
- DZHK (German Center for Cardiovascular Research)Partner Site RhineMainMainzGermany
- Institute for Clinical Chemistry and Laboratory MedicineUniversity Medical Center of the Johannes Gutenberg‐University MainzMainzGermany
| | - Hugo ten Cate
- Laboratory for Clinical Thrombosis and Hemostasis, Department of Internal Medicine, Cardiovascular Research Institute Maastricht (CARIM)Maastricht University Medical CenterMaastrichtThe Netherlands
- Center for Thrombosis and Hemostasis (CTH)University Medical Center of the Johannes Gutenberg‐University MainzMainzGermany
| | - Philipp S. Wild
- Center for Thrombosis and Hemostasis (CTH)University Medical Center of the Johannes Gutenberg‐University MainzMainzGermany
- DZHK (German Center for Cardiovascular Research)Partner Site RhineMainMainzGermany
- Preventive Cardiology and Preventive Medicine, Center for CardiologyUniversity Medical Center of the Johannes Gutenberg‐University MainzMainzGermany
| | - Henri M. H. Spronk
- Laboratory for Clinical Thrombosis and Hemostasis, Department of Internal Medicine, Cardiovascular Research Institute Maastricht (CARIM)Maastricht University Medical CenterMaastrichtThe Netherlands
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La Mura V, Gagliano N, Arnaboldi F, Sartori P, Procacci P, Denti L, Liguori E, Bitto N, Ristagno G, Latini R, Dondossola D, Salerno F, Tripodi A, Colombo M, Peyvandi F. Simvastatin Prevents Liver Microthrombosis and Sepsis Induced Coagulopathy in a Rat Model of Endotoxemia. Cells 2022; 11:1148. [PMID: 35406712 PMCID: PMC8997834 DOI: 10.3390/cells11071148] [Citation(s) in RCA: 11] [Impact Index Per Article: 3.7] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/18/2022] [Revised: 03/19/2022] [Accepted: 03/22/2022] [Indexed: 01/27/2023] Open
Abstract
Background: Endotoxemia causes endothelial dysfunction and microthrombosis, which are pathogenic mechanisms of coagulopathy and organ failure during sepsis. Simvastatin has potential anti-thrombotic effects on liver endothelial cells. We investigated the hemostatic changes induced by lipopolysaccharide (LPS) and explored the protective effects of simvastatin against liver vascular microthrombosis. Methods and results: We compared male Wistar rats exposed to LPS (5 mg/kg one i.p. dose) or saline in two experimental protocols—placebo (vehicle) and simvastatin (25 mg/kg die, orally, for 3 days before LPS). Morphological studies were performed by light- and electron-microscopy analyses to show intravascular fibrin deposition, vascular endothelial structure and liver damage. Peripheral- and organ-hemostatic profiles were analyzed using whole blood viscoelastometry by ROTEM, liver biopsy and western-blot/immunohistochemistry of thrombomodulin (TM), as well as immunohistochemistry of the von Willebrand factor (VWF). LPS-induced fibrin deposition and liver vascular microthrombosis were combined with a loss of sinusoidal endothelial TM expression and VWF-release. These changes were associated with parenchymal eosinophilia and necrosis. ROTEM analyses displayed hypo-coagulability in the peripheral blood that correlated with the degree of intrahepatic fibrin deposition (p < 0.05). Simvastatin prevented LPS-induced fibrin deposition by preserving TM expression in sinusoidal cells and completely reverted the peripheral hypo-coagulability caused by endotoxemia. These changes were associated with a significant reduction of liver cell necrosis without any effect on eosinophilia. Conclusions: Simvastatin preserves the antithrombotic properties of sinusoidal endothelial cells disrupted by LPS, deserving pharmacological properties to contrast sepsis-associated coagulopathy and hepatic failure elicited by endotoxemia
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Affiliation(s)
- Vincenzo La Mura
- Fondazione I.R.C.C.S. Ca’ Granda, Ospedale Maggiore Policlinico, U.O.C. Medicina Generale Emostasi e Trombosi, 20122 Milan, Italy; (N.B.); (A.T.); (F.P.)
- CRC “A.M. e A. Migliavacca” per lo Studio e la Cura delle Malattie del Fegato, Università degli Studi di Milano, 20122 Milan, Italy
- Dipartimento di Fisiopatologia dei Trapianti, Università degli Studi di Milano, 20132 Milan, Italy; (E.L.); (G.R.); (D.D.)
| | - Nicoletta Gagliano
- Dipartimento di Scienze Biomediche per la Salute, Università degli Studi di Milano, 20133 Milan, Italy; (N.G.); (F.A.); (P.S.); (P.P.); (L.D.); (F.S.)
| | - Francesca Arnaboldi
- Dipartimento di Scienze Biomediche per la Salute, Università degli Studi di Milano, 20133 Milan, Italy; (N.G.); (F.A.); (P.S.); (P.P.); (L.D.); (F.S.)
| | - Patrizia Sartori
- Dipartimento di Scienze Biomediche per la Salute, Università degli Studi di Milano, 20133 Milan, Italy; (N.G.); (F.A.); (P.S.); (P.P.); (L.D.); (F.S.)
| | - Patrizia Procacci
- Dipartimento di Scienze Biomediche per la Salute, Università degli Studi di Milano, 20133 Milan, Italy; (N.G.); (F.A.); (P.S.); (P.P.); (L.D.); (F.S.)
| | - Luca Denti
- Dipartimento di Scienze Biomediche per la Salute, Università degli Studi di Milano, 20133 Milan, Italy; (N.G.); (F.A.); (P.S.); (P.P.); (L.D.); (F.S.)
| | - Eleonora Liguori
- Dipartimento di Fisiopatologia dei Trapianti, Università degli Studi di Milano, 20132 Milan, Italy; (E.L.); (G.R.); (D.D.)
| | - Niccolò Bitto
- Fondazione I.R.C.C.S. Ca’ Granda, Ospedale Maggiore Policlinico, U.O.C. Medicina Generale Emostasi e Trombosi, 20122 Milan, Italy; (N.B.); (A.T.); (F.P.)
| | - Giuseppe Ristagno
- Dipartimento di Fisiopatologia dei Trapianti, Università degli Studi di Milano, 20132 Milan, Italy; (E.L.); (G.R.); (D.D.)
- Fondazione I.R.C.C.S. Ca’ Granda, Ospedale Maggiore Policlinico, U.O.C. Anestesia e Rianimazione, 20122 Milan, Italy
| | - Roberto Latini
- Dipartimento di Ricerca Cardiovascolare, Istituto di Ricerche Farmacologiche Mario Negri I.R.C.C.S., 20156 Milan, Italy;
| | - Daniele Dondossola
- Dipartimento di Fisiopatologia dei Trapianti, Università degli Studi di Milano, 20132 Milan, Italy; (E.L.); (G.R.); (D.D.)
- U.O. Chirurgia Generale e dei Trapianti di Fegato, Fondazione IRCCS Ca′ Granda Ospedale Maggiore Policlinico, 20122 Milan, Italy
| | - Francesco Salerno
- Dipartimento di Scienze Biomediche per la Salute, Università degli Studi di Milano, 20133 Milan, Italy; (N.G.); (F.A.); (P.S.); (P.P.); (L.D.); (F.S.)
| | - Armando Tripodi
- Fondazione I.R.C.C.S. Ca’ Granda, Ospedale Maggiore Policlinico, U.O.C. Medicina Generale Emostasi e Trombosi, 20122 Milan, Italy; (N.B.); (A.T.); (F.P.)
| | - Massimo Colombo
- Liver Center IRCCS San Raffaele Hospital, 20132 Milan, Italy;
| | - Flora Peyvandi
- Fondazione I.R.C.C.S. Ca’ Granda, Ospedale Maggiore Policlinico, U.O.C. Medicina Generale Emostasi e Trombosi, 20122 Milan, Italy; (N.B.); (A.T.); (F.P.)
- Dipartimento di Fisiopatologia dei Trapianti, Università degli Studi di Milano, 20132 Milan, Italy; (E.L.); (G.R.); (D.D.)
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11
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Gowani F, Phillips B, Leveque C, Castillo B, Chen J, Chandler W, Rice L, Salazar E. Recurrent Gastrointestinal Bleeding in a Middle-Aged Man. Lab Med 2021; 53:e91-e94. [DOI: 10.1093/labmed/lmab104] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/12/2022] Open
Abstract
Abstract
Acquired von Willebrand disease (avWD) arises because of mechanisms that destroy, decrease, absorb, or clear von Willebrand factor (vWF). A 59-year-old man presented with a 3-year history of recurrent gastrointestinal bleeding. Laboratory workup revealed a prolonged platelet function assay-100. The vWF antigen was decreased, and a low vWF immunofunctional activity/antigen ratio, low collagen binding/antigen ratio, and decreased intermediate and high molecular weight multimers were noted. The patient had no high-shear stress conditions, and an antibody-mediated process was suspected. A vWF mixing study showed complete correction of vWF activity, suggesting no direct functional inhibitor. The patient was given a bolus of vWF concentrate with serial measurements of vWF; the vWF half-life was 2.5 hours. The vWF propeptide/antigen ratio was 4:1, supporting a diagnosis of aVWD resulting from increased antibody-mediated vWF clearance. This case study emphasizes the laboratory’s role in the diagnosis and treatment of rare, overlooked acquired bleeding disorders.
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Affiliation(s)
- Faaria Gowani
- University of Tennessee Health Science Center, Department of Pathology and Laboratory Medicine, Memphis, Tennessee, US
| | - Bonnie Phillips
- Houston Methodist Hospital, Department of Pathology and Genomic Medicine, Houston, Texas, US
| | - Christopher Leveque
- Houston Methodist Hospital, Department of Pathology and Genomic Medicine, Houston, Texas, US
| | - Brian Castillo
- Houston Methodist Hospital, Department of Pathology and Genomic Medicine, Houston, Texas, US
| | - Jian Chen
- Houston Methodist Hospital, Department of Pathology and Genomic Medicine, Houston, Texas, US
| | - Wayne Chandler
- Seattle Children’s Hospital, Department of Laboratories, Seattle, Washington, US
| | - Lawrence Rice
- Houston Methodist Hospital, Department of Medicine and Cancer Center, Houston, Texas, US
| | - Eric Salazar
- Houston Methodist Hospital, Department of Pathology and Genomic Medicine, Houston, Texas, US
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12
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Häußler KS, Keese M, Weber CF, Geisen C, Miesbach W. Prospective Evaluation of the Pre-, Intra-, and Postoperative Kinetics of ADAMTS-13, von Willebrand Factor, and Interleukin-6 in Vascular Surgery. Clin Appl Thromb Hemost 2021; 26:1076029620930273. [PMID: 33023308 PMCID: PMC7545751 DOI: 10.1177/1076029620930273] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/16/2022] Open
Abstract
Postoperative thrombotic thrombocytopenic purpura (TTP) shows clinical presentation similar to classical TTP, whereas exact pathophysiological contexts remain unexplained. In this study, we investigated intraoperative and postoperative changes in ADAMTS-13 (a disintegrin and metalloprotease with thrombospondin type 1 motifs, member 13), von Willebrand factor (VWF), large VWF multimers, and interleukin-6 (IL-6) in vascular surgery patients. The objective was to compare the impact of endovascular, peripheral, and aortic surgery on target parameters which are supposed to play a role in surgery-associated TTP. A total of 93 vascular surgery patients were included and divided into 4 groups according to the specific type of intervention they underwent. Blood samples were taken preoperatively, intraoperatively, and postoperatively on days 2 and 4. The ADAMTS-13 activity decreased significantly in 3 of the 4 groups during surgery (from median 81% to 49%, P < .001, in the group undergoing aortoiliacal interventions), whereas the percentage of large VWF multimers increased in all groups of patients. von Willebrand factor antigen increased significantly in all groups on postoperative day 2 and IL-6 increased significantly in the intraoperative and early postoperative period. There was no significant correlation between the intraoperative decrease in ADAMTS-13 and the increase in VWF or IL-6. No patient in this study showed clinical picture of TTP; the precise cause and clinical significance of moderately reduced ADAMTS-13 activity in the perioperative setting have not yet been definitely determined.
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Affiliation(s)
- Katja Susanne Häußler
- University Hospital Frankfurt, Medical Clinic II, Institute of Transfusion Medicine and Immunohematology, Hemophilia Center, Frankfurt, Germany
| | - Michael Keese
- Department for Vascular Surgery, University Hospital Mannheim, Mannheim, Germany
| | - Christian Friedrich Weber
- Department for Anesthesiology, Intensive Care Medicine and Emergency Medicine, Asklepios Klinik Wandsbek, Hamburg, Germany
| | - Christof Geisen
- German Red Cross Blood Center Frankfurt am Main, Institute of Transfusion Medicine and Immunohematology, Frankfurt, Germany
| | - Wolfgang Miesbach
- University Hospital Frankfurt, Medical Clinic II, Institute of Transfusion Medicine and Immunohematology, Hemophilia Center, Frankfurt, Germany
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13
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O'Brien HER, Zhang XF, Sanz-Hernandez M, Chion A, Shapiro S, Mobayen G, Xu Y, De Simone A, Laffan MA, McKinnon TAJ. Blocking von Willebrand factor free thiols inhibits binding to collagen under high and pathological shear stress. J Thromb Haemost 2021; 19:358-369. [PMID: 33075181 DOI: 10.1111/jth.15142] [Citation(s) in RCA: 4] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/07/2020] [Revised: 10/09/2020] [Accepted: 10/12/2020] [Indexed: 01/04/2023]
Abstract
BACKGROUND Von Willebrand factor (VWF) contains a number of free thiols, the majority of which are located in its C-domains, and these have been shown to alter VWF function, However, the impact of free thiols on function following acute exposure of VWF to collagen under high and pathological shear stress has not been determined. METHODS VWF free thiols were blocked with N-ethylmaleimide and flow assays performed under high and pathological shear rates to determine the impact on platelet capture and collagen binding function. Atomic force microscopy (AFM) was used to probe the interaction of VWF with collagen and molecular simulations conducted to determine the effect of free thiols on the flexibility of the VWF-C4 domain. RESULTS Blockade of VWF free thiols reduced VWF-mediated platelet capture to collagen in a shear-dependent manner, with platelet capture virtually abolished above 5000 s-1 and in regions of stenosis in microfluidic channels. Direct visualization of VWF fibers formed under extreme pathological shear rates and analysis of collagen-bound VWF attributed the effect to altered binding of VWF to collagen. AFM measurements showed that thiol-blockade reduced the lifetime and strength of the VWF-collagen bond. Pulling simulations of the VWF-C4 domain demonstrated that with one or two reduced disulphide bonds the C4 domain has increased flexibility and the propensity to undergo free-thiol exchange. CONCLUSIONS We conclude that free thiols in the C-domains of VWF enhance the flexibility of the molecule and enable it to withstand high shear forces following collagen binding, demonstrating a previously unrecognized role for VWF free thiols.
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Affiliation(s)
- Harrison E R O'Brien
- Department of Immunology and Inflammation, Centre for Haematology, Imperial College of Science Technology and Medicine, London, UK
- Institute of Structural and Molecular Biology, University College London, London, UK
| | - X Frank Zhang
- Department of Bioengineering, Department of Mechanical Engineering and Mechanics, Lehigh University, Bethlehem, PA, USA
- Department of Mechanical Engineering and Mechanics, Lehigh University, Bethlehem, PA, USA
| | | | - Alain Chion
- Irish Centre for Vascular Biology, Molecular and Cellular Therapeutics, Royal College of Surgeons in Ireland, Dublin, Ireland
| | - Susan Shapiro
- Oxford University Hospitals NHS Foundation Trust, Oxford NIHR Biomedical Research Centre, Oxford, UK
- Radcliffe Department of Medicine, Oxford University, Oxford, UK
| | - Golzar Mobayen
- Department of Immunology and Inflammation, Centre for Haematology, Imperial College of Science Technology and Medicine, London, UK
| | - Yan Xu
- Department of Bioengineering, Department of Mechanical Engineering and Mechanics, Lehigh University, Bethlehem, PA, USA
- Department of Mechanical Engineering and Mechanics, Lehigh University, Bethlehem, PA, USA
| | | | - Michael A Laffan
- Department of Immunology and Inflammation, Centre for Haematology, Imperial College of Science Technology and Medicine, London, UK
| | - Thomas A J McKinnon
- Department of Immunology and Inflammation, Centre for Haematology, Imperial College of Science Technology and Medicine, London, UK
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14
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Belyaev AV. Intradimer forces and their implication for conformations of von Willebrand factor multimers. Biophys J 2021; 120:899-911. [PMID: 33524374 DOI: 10.1016/j.bpj.2021.01.022] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/09/2020] [Revised: 12/31/2020] [Accepted: 01/19/2021] [Indexed: 10/22/2022] Open
Abstract
The largest blood glycoprotein von Willebrand factor (VWF) responds to hydrodynamic stresses in the bloodstream with abrupt conformation changes, thus increasing its adhesivity to platelets and collagen. Arterial and microvascular hemostasis relies on mechanical and physicochemical properties of this macromolecule. Recently, it was discovered that the mechanical properties of VWF are controlled by multiple pH-dependent interactions with opposite trends within dimeric subunits. In this work, computer simulations reveal the effect of these intradimer forces on the conformation of VWF multimers in various hydrodynamic conditions. A coarse-grained computer model of VWF has been proposed and parameterized to give a good agreement with experimental data. The simulations suggest that strong attraction between VWF D4 domains increases the resistance to elongation under shear stress, whereas even intermediate attraction between VWF C domains contributes to VWF compaction in nonsheared fluid. It is hypothesized that the detailed subdimer dynamics of VWF concatamers may be one of the biophysical regulators of initial hemostasis and arterial thrombosis.
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Affiliation(s)
- Aleksey V Belyaev
- Lomonosov Moscow State University, Faculty of Physics, Moscow, Russia; IRC Mathematical modelling in Biomedicine, S.M. Nikolskii Mathematical Institute, RUDN University, Moscow, Russia.
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15
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Zhu JJ, Jiang ZT, Liu C, Xi YF, Wang J, Yang FF, Yao WJ, Pang W, Han LL, Zhang YH, Sun AQ, Zhou J. VAMP3 and SNAP23 as Potential Targets for Preventing the Disturbed Flow-Accelerated Thrombus Formation. Front Cell Dev Biol 2020; 8:576826. [PMID: 33224946 PMCID: PMC7674309 DOI: 10.3389/fcell.2020.576826] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/27/2020] [Accepted: 10/05/2020] [Indexed: 01/21/2023] Open
Abstract
Disturbed blood flow has been recognized to promote platelet aggregation and thrombosis via increasing accumulation of von Willebrand factor (VWF) at the arterial post-stenotic sites. The mechanism underlying the disturbed-flow regulated endothelial VWF production remains elusive. Here we described a mouse model, in which the left external carotid artery (LECA) is ligated to generate disturbed flow in the common carotid artery. Ligation of LECA increased VWF accumulation in the plasma. Carotid arterial thrombosis was induced by ferric chloride (FeCl3) application and the time to occlusion in the ligated vessels was reduced in comparison with the unligated vessels. In vitro, endothelial cells were subjected to oscillatory shear (OS, 0.5 ± 4 dynes/cm2) or pulsatile shear (PS, 12 ± 4 dynes/cm2). OS promoted VWF secretion as well as the cell conditioned media-induced platelet aggregation by regulating the intracellular localization of vesicle-associated membrane protein 3 (VAMP3) and synaptosomal-associated protein 23 (SNAP23). Disruption of vimentin intermediate filaments abolished the OS-induced translocation of SNAP23 to the cell membrane. Knockdown of VAMP3 and SNAP23 reduced the endothelial secretion of VWF. Systemic inhibition of VAMP3 and SNAP23 by treatment of mice with rapamycin significantly ameliorated the FeCl3-induced thrombogenesis, whereas intraluminal overexpression of VAMP3 and SNAP23 aggravated it. Our findings demonstrate VAMP3 and SNAP23 as potential targets for preventing the disturbed flow-accelerated thrombus formation.
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Affiliation(s)
- Juan-Juan Zhu
- Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Peking University, Beijing, China.,Key Laboratory of Molecular Cardiovascular Sciences, Ministry of Education, Beijing, China.,National Health Commission of the People's Republic of China Key Laboratory of Cardiovascular Molecular Biology and Regulatory Peptides, Peking University, Beijing, China.,Department of Pharmacology, School of Basic Medical Science, Peking University, Beijing, China
| | - Zhi-Tong Jiang
- Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Peking University, Beijing, China.,Key Laboratory of Molecular Cardiovascular Sciences, Ministry of Education, Beijing, China.,National Health Commission of the People's Republic of China Key Laboratory of Cardiovascular Molecular Biology and Regulatory Peptides, Peking University, Beijing, China
| | - Chen Liu
- Department of Clinical Laboratory, Peking University People's Hospital, Beijing, China
| | - Yi-Feng Xi
- School of Biological Science and Medical Engineering, Beijing Advanced Innovation Center for Biomedical Engineering, Beihang University, Beijing, China
| | - Jin Wang
- Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Peking University, Beijing, China.,Key Laboratory of Molecular Cardiovascular Sciences, Ministry of Education, Beijing, China.,National Health Commission of the People's Republic of China Key Laboratory of Cardiovascular Molecular Biology and Regulatory Peptides, Peking University, Beijing, China
| | - Fang-Fang Yang
- Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Peking University, Beijing, China.,Key Laboratory of Molecular Cardiovascular Sciences, Ministry of Education, Beijing, China.,National Health Commission of the People's Republic of China Key Laboratory of Cardiovascular Molecular Biology and Regulatory Peptides, Peking University, Beijing, China
| | - Wei-Juan Yao
- Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Peking University, Beijing, China
| | - Wei Pang
- Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Peking University, Beijing, China
| | - Li-Li Han
- Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Peking University, Beijing, China
| | - Yong-He Zhang
- Department of Pharmacology, School of Basic Medical Science, Peking University, Beijing, China
| | - An-Qiang Sun
- School of Biological Science and Medical Engineering, Beijing Advanced Innovation Center for Biomedical Engineering, Beihang University, Beijing, China
| | - Jing Zhou
- Department of Physiology and Pathophysiology, School of Basic Medical Sciences, Peking University, Beijing, China.,Key Laboratory of Molecular Cardiovascular Sciences, Ministry of Education, Beijing, China.,National Health Commission of the People's Republic of China Key Laboratory of Cardiovascular Molecular Biology and Regulatory Peptides, Peking University, Beijing, China
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16
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Schneider MF, Fallah MA, Mess C, Obser T, Schneppenheim R, Alexander-Katz A, Schneider SW, Huck V. Platelet adhesion and aggregate formation controlled by immobilised and soluble VWF. BMC Mol Cell Biol 2020; 21:64. [PMID: 32917131 PMCID: PMC7488753 DOI: 10.1186/s12860-020-00309-7] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/23/2020] [Accepted: 08/26/2020] [Indexed: 11/21/2022] Open
Abstract
Background It has been demonstrated that von Willebrand factor (VWF) mediated platelet-endothelium and platelet-platelet interactions are shear dependent. The VWF’s mobility under dynamic conditions (e.g. flow) is pivotal to platelet adhesion and VWF-mediated aggregate formation in the cascade of VWF-platelet interactions in haemostasis. Results Combining microfluidic tools with fluorescence and reflection interference contrast microscopy (RICM), here we show, that specific deletions in the A-domains of the biopolymer VWF affect both, adhesion and aggregation properties independently. Intuitively, the deletion of the A1-domain led to a significant decrease in both adhesion and aggregate formation of platelets. Nevertheless, the deletion of the A2-domain revealed a completely different picture, with a significant increase in formation of rolling aggregates (gain of function). We predict that the A2-domain effectively ‘masks’ the potential between the platelet glycoprotein (GP) Ib and the VWF A1-domain. Furthermore, the deletion of the A3-domain led to no significant variation in either of the two functional characteristics. Conclusions These data demonstrate that the macroscopic functional properties i.e. adhesion and aggregate formation cannot simply be assigned to the properties of one particular domain, but have to be explained by cooperative phenomena. The absence or presence of molecular entities likewise affects the properties (thermodynamic phenomenology) of its neighbours, therefore altering the macromolecular function.
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Affiliation(s)
- Matthias F Schneider
- Department of Physics, Medical and Biological Physics, Technical University Dortmund, Emil-Figge-Str. 50, 44227, Dortmund, Germany
| | - Mohammad A Fallah
- Department of Chemistry, University of Konstanz, Universitätsstr. 10, 78457, Constance, Germany
| | - Christian Mess
- University Medical Centre Hamburg-Eppendorf, Centre for Internal Medicine, Martinistr. 52, 20246, Hamburg, Germany
| | - Tobias Obser
- Department of Paediatric Haematology and Oncology, University Medical Centre Hamburg-Eppendorf, Martinistr. 52, 20246, Hamburg, Germany
| | - Reinhard Schneppenheim
- Department of Paediatric Haematology and Oncology, University Medical Centre Hamburg-Eppendorf, Martinistr. 52, 20246, Hamburg, Germany
| | - Alfredo Alexander-Katz
- Department of Materials Science and Engineerin, Massachusetts Institute of Technology, 400 Technology Sq. (NE46-605), Cambridge, MA, 02139, USA
| | - Stefan W Schneider
- University Medical Centre Hamburg-Eppendorf, Centre for Internal Medicine, Martinistr. 52, 20246, Hamburg, Germany
| | - Volker Huck
- University Medical Centre Hamburg-Eppendorf, Centre for Internal Medicine, Martinistr. 52, 20246, Hamburg, Germany. .,Heidelberg University, Medical Faculty Mannheim, Experimental Dermatology, Theodor-Kutzer-Ufer 1-3, 68167, Mannheim, Germany.
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17
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Steinert M, Ramming I, Bergmann S. Impact of Von Willebrand Factor on Bacterial Pathogenesis. Front Med (Lausanne) 2020; 7:543. [PMID: 33015097 PMCID: PMC7494747 DOI: 10.3389/fmed.2020.00543] [Citation(s) in RCA: 21] [Impact Index Per Article: 4.2] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/28/2020] [Accepted: 07/30/2020] [Indexed: 12/31/2022] Open
Abstract
Von Willebrand factor (VWF) is a mechano-sensitive protein with crucial functions in normal hemostasis, which are strongly dependant on the shear-stress mediated defolding and multimerization of VWF in the blood stream. Apart from bleeding disorders, higher plasma levels of VWF are often associated with a higher risk of cardiovascular diseases. Herein, the disease symptoms are attributed to the inflammatory response of the activated endothelium and share high similarities to the reaction of the host vasculature to systemic infections caused by pathogenic bacteria such as Staphylococcus aureus and Streptococcus pneumoniae. The bacteria recruit circulating VWF, and by binding to immobilized VWF on activated endothelial cells in blood flow, they interfere with the physiological functions of VWF, including platelet recruitment and coagulation. Several bacterial VWF binding proteins have been identified and further characterized by biochemical analyses. Moreover, the development of a combination of sophisticated cell culture systems simulating shear stress levels of the blood flow with microscopic visualization also provided valuable insights into the interaction mechanism between bacteria and VWF-strings. In vivo studies using mouse models of bacterial infection and zebrafish larvae provided evidence that the interaction between bacteria and VWF promotes bacterial attachment, coagulation, and thrombus formation, and thereby contributes to the pathophysiology of severe infectious diseases such as infective endocarditis and bacterial sepsis. This mini-review summarizes the current knowledge of the interaction between bacteria and the mechano-responsive VWF, and corresponding pathophysiological disease symptoms.
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Affiliation(s)
- Michael Steinert
- Institut für Mikrobiologie, Technische Universität Braunschweig, Braunschweig, Germany.,Department of Infection Biology, Helmholtz Center for Infection Diseases, Braunschweig, Germany
| | - Isabell Ramming
- Institut für Mikrobiologie, Technische Universität Braunschweig, Braunschweig, Germany
| | - Simone Bergmann
- Institut für Mikrobiologie, Technische Universität Braunschweig, Braunschweig, Germany
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18
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Grzegorski SJ, Hu Z, Liu Y, Yu X, Ferguson AC, Madarati H, Friedmann AP, Reyon D, Kim PY, Kretz CA, Joung JK, Shavit JA. Disruption of the kringle 1 domain of prothrombin leads to late onset mortality in zebrafish. Sci Rep 2020; 10:4049. [PMID: 32132579 PMCID: PMC7055286 DOI: 10.1038/s41598-020-60840-7] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Grants] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 10/01/2019] [Accepted: 01/22/2020] [Indexed: 02/04/2023] Open
Abstract
The ability to prevent blood loss in response to injury is a conserved function of all vertebrates. Complete deficiency of the central clotting enzyme prothrombin has never been observed in humans and is incompatible with postnatal life in mice, thus limiting the ability to study its role in vivo. Zebrafish are able to tolerate severe hemostatic deficiencies that are lethal in mammals. We have generated a targeted genetic deletion in the kringle 1 domain of zebrafish prothrombin. Homozygous mutant embryos develop normally into the mid-juvenile stage but demonstrate complete mortality by 2 months of age primarily due to internal hemorrhage. Mutants are unable to form occlusive venous and arterial thrombi in response to endothelial injury, a defect that was phenocopied using direct oral anticoagulants. Human prothrombin engineered with the equivalent mutation exhibits a severe reduction in secretion, thrombin generation, and fibrinogen cleavage. Together, these data demonstrate the conserved function of thrombin in zebrafish and provide insight into the role of kringle 1 in prothrombin maturation and activity. Understanding how zebrafish are able to develop normally and survive into early adulthood without thrombin activity will provide important insight into its pleiotropic functions as well as the management of patients with bleeding disorders.
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Affiliation(s)
| | - Zhilian Hu
- Department of Pediatrics, University of Michigan, Ann Arbor, MI, USA
- Oxford University, Oxford, UK
| | - Yang Liu
- Department of Pediatrics, University of Michigan, Ann Arbor, MI, USA
- Molecular Innovations, Inc., Novi, MI, USA
| | - Xinge Yu
- Department of Pediatrics, University of Michigan, Ann Arbor, MI, USA
| | | | - Hasam Madarati
- Department of Medicine, McMaster University, Hamilton, ON, Canada
- Thromosis and Atherosclerosis Research Institute, Hamilton, ON, Canada
| | - Alexander P Friedmann
- Department of Medicine, McMaster University, Hamilton, ON, Canada
- Thromosis and Atherosclerosis Research Institute, Hamilton, ON, Canada
| | - Deepak Reyon
- Molecular Pathology Unit, Massachusetts General Hospital, Charlestown, MA, USA
- Department of Pathology, Harvard Medical School, Boston, MA, USA
- Editas Medicine Inc., Cambridge, MA, USA
| | - Paul Y Kim
- Department of Medicine, McMaster University, Hamilton, ON, Canada
- Thromosis and Atherosclerosis Research Institute, Hamilton, ON, Canada
| | - Colin A Kretz
- Department of Medicine, McMaster University, Hamilton, ON, Canada
- Thromosis and Atherosclerosis Research Institute, Hamilton, ON, Canada
| | - J Keith Joung
- Molecular Pathology Unit, Massachusetts General Hospital, Charlestown, MA, USA
- Department of Pathology, Harvard Medical School, Boston, MA, USA
| | - Jordan A Shavit
- Department of Pediatrics, University of Michigan, Ann Arbor, MI, USA.
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19
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Rasmussen KL, Philips M, Tripodi A, Goetze JP. Unexpected, isolated activated partial thromboplastin time prolongation: A practical mini-review. Eur J Haematol 2020; 104:519-525. [PMID: 32049377 DOI: 10.1111/ejh.13394] [Citation(s) in RCA: 13] [Impact Index Per Article: 2.6] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/27/2019] [Revised: 02/07/2020] [Accepted: 02/07/2020] [Indexed: 01/03/2023]
Abstract
A common inquiry in coagulation laboratories is how to interpret an unexpected, isolated prolonged activated partial thromboplastin time (APTT). In this context, isolated means together with a normal prothrombin time (PT) and/or normal international normalized ratio (INR). This finding may lead to contact with laboratory doctors for further advice on a diagnostic strategy. Occasionally, the need for a diagnostic algorithm can be subacute, where surgery has to be postponed until an explanation for the isolated, prolonged APTT has been established. Activated partial thromboplastin time as a coagulation test was developed to monitor patients with hemophilia. Different APTT reagents display considerable differences in their sensitivity to deficiencies of coagulation factors. An isolated, prolonged APTT is seen in (a) individuals/patients with lupus anticoagulants, (b) patients in treatment with anticoagulants, mainly heparin, and (c) patients with deficiencies of specific coagulation factors. In this tutorial review, we summarize what may cause an isolated prolonged APTT and we present a simple diagnostic algorithm to differentiate between lupus anticoagulants (common) and factor deficiencies (rare). The identification of an isolated prolonged APTT as well as the underlying cause can be of the utmost importance in ensuring the correct therapeutic follow-up.
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Affiliation(s)
| | - Malou Philips
- Department of Clinical Biochemistry, Rigshospitalet, Copenhagen, Denmark
| | - Armando Tripodi
- Fondazione IRCCS Cà Granda Ospedale Maggiore Policlinico, Angelo Bianchi Bonomi Hemophilia and Thrombosis Center and Fondazione Luigi Villa, Milano, Italy
| | - Jens Peter Goetze
- Department of Clinical Biochemistry, Rigshospitalet, Copenhagen, Denmark.,Department of Biomedical Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark
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20
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Ngo T, Kim K, Bian Y, Nam G, Park HJ, Lee K, Cho GS, Ryu JM, Lim KM, Chung JH. Antithrombotic effect of SP-8008, a benzoic acid derivative, through the selective inhibition of shear stress-induced platelet aggregation. Br J Pharmacol 2019; 177:929-944. [PMID: 31648364 PMCID: PMC7024737 DOI: 10.1111/bph.14894] [Citation(s) in RCA: 6] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 03/30/2019] [Revised: 07/30/2019] [Accepted: 09/05/2019] [Indexed: 01/23/2023] Open
Abstract
Background and Purpose Bleeding is one of the most critical adverse effects of antithrombotic drugs, and many efforts have been made to discover novel antiplatelet agents without bleeding complications. Shear stress‐induced platelet aggregation (SIPA), where the interaction of von Willebrand factor (vWF) and platelet glycoprotein (GP) Ib constitutes the initial step, is a promising target to overcome bleeding problems, as SIPA occurs only in pathological conditions. Here, we describe SP‐8008, a novel modulator of vWF–GP Ib interactions and evaluated its antiplatelet/antithrombotic effects. Experimental Approach Newly synthesized compounds were screened for antiplatelet effects in vitro, using human platelets exposed to high shear stress. Aggregation, intracellular calcium level, granule secretion, and integrin activation were assessed. Molecular modelling using virtual docking and flow cytometry were used to evaluate effects on vWF–GP Ib interactions. Antithrombotic effects in vivo were determined in rats, using arterial thrombosis and shear stress‐specific thrombosis. Transection tail bleeding time was used to evaluate adverse effects. Key Results SP‐8008 was a potent inhibitor of SIPA, with IC50 of 1.44 ± 0.09 μM. SP‐8008 effectively and broadly blocked shear stress‐induced platelet activation events, without any significant toxicity. Importantly, SP‐8008 was highly selective against SIPA, effectively interfering with vWF–GP Ib engagement. Most importantly, SP‐8008 exerted significant antithrombotic effects in vivo in both shear stress‐specific and arterial thrombosis, without prolonging bleeding time. Conclusions and Implications Our results demonstrated that SP‐8008 can be a novel selective antiplatelet agent with improved safety profile.
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Affiliation(s)
- Thien Ngo
- College of Pharmacy, Seoul National University, Seoul, Korea
| | - Keunyoung Kim
- College of Pharmacy, Seoul National University, Seoul, Korea
| | - Yiying Bian
- College of Pharmacy, Seoul National University, Seoul, Korea
| | - Gibeom Nam
- School of Pharmacy, Sungkyunkwan University, Suwon, Korea
| | - Hyun-Ju Park
- School of Pharmacy, Sungkyunkwan University, Suwon, Korea
| | - Kiho Lee
- College of Pharmacy, Korea University, Sejong, Korea
| | - Geum-Sil Cho
- Research Headquarters, Shin Poong Pharm. Co. Ltd., Ansan, Korea
| | - Jei-Man Ryu
- Research Headquarters, Shin Poong Pharm. Co. Ltd., Ansan, Korea
| | - Kyung-Min Lim
- College of Pharmacy, Ewha Womans University, Seoul, Korea
| | - Jin-Ho Chung
- College of Pharmacy, Seoul National University, Seoul, Korea
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21
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Kushchenko YK, Belyaev AV. Effects of hydrophobicity, tethering and size on flow-induced activation of von Willebrand factor multimers. J Theor Biol 2019; 485:110050. [PMID: 31618612 DOI: 10.1016/j.jtbi.2019.110050] [Citation(s) in RCA: 12] [Impact Index Per Article: 2.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/26/2019] [Revised: 09/12/2019] [Accepted: 10/12/2019] [Indexed: 01/14/2023]
Abstract
Von Willebrand factor (VWF) is a multimeric protein of blood plasma that mediates platelet adhesion to injury under strong hemodynamic flows in arterias and microvasvulature. We present a 3D coarse-grained computer model of VWF multimers in flowing viscous fluid that explicitely grasps the dynamics, the conformational changes and the hydrodynamics-induced activation of adhesivity of these protein concatamers to GPIb receptor of blood platelets. The model is based on the fluctuating Lattice Boltzmann method for modelling the hydrodynamics in the simulation box and the Lagrangian particle dynamics coupled to the fluid by a viscous drag force. The model has been validated by the comparison with the experimental data found in literature. We studied the effect of hydrophobic interactions on the conformational dynamics of VWF multimers. The simulations suggest that the contour length is an important parameter that controls the functionality of VWF multimers in blood. We also demonstrate that tethering to the surface of a vessel wall promoted the flow-induced activation of VWF, while those multimers that remain untethered and move freely in the blood plasma require a stronger shearing to get activated.
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Affiliation(s)
- Yulia K Kushchenko
- Lomonosov Moscow State University, Faculty of Physics, Moscow 119991, Russia
| | - Aleksey V Belyaev
- Lomonosov Moscow State University, Faculty of Physics, Moscow 119991, Russia; S.M. Nikol'skii Mathematical Institute, RUDN University, Moscow 115419, Russia.
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22
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Rana A, Westein E, Niego B, Hagemeyer CE. Shear-Dependent Platelet Aggregation: Mechanisms and Therapeutic Opportunities. Front Cardiovasc Med 2019; 6:141. [PMID: 31620451 PMCID: PMC6763557 DOI: 10.3389/fcvm.2019.00141] [Citation(s) in RCA: 120] [Impact Index Per Article: 20.0] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/05/2019] [Accepted: 09/03/2019] [Indexed: 01/04/2023] Open
Abstract
Cardiovascular diseases (CVD) are the number one cause of morbidity and death worldwide. As estimated by the WHO, the global death rate from CVD is 31% wherein, a staggering 85% results from stroke and myocardial infarction. Platelets, one of the key components of thrombi, have been well-investigated over decades for their pivotal role in thrombus development in healthy as well as diseased blood vessels. In hemostasis, when a vascular injury occurs, circulating platelets are arrested at the site of damage, where they are activated and aggregate to form hemostatic thrombi, thus preventing further bleeding. However, in thrombosis, pathological activation of platelets occurs, leading to uncontrolled growth of a thrombus, which in turn can occlude the blood vessel or embolize, causing downstream ischemic events. The molecular processes causing pathological thrombus development are in large similar to the processes controlling physiological thrombus formation. The biggest challenge of anti-thrombotics and anti-platelet therapeutics has been to decouple the pathological platelet response from the physiological one. Currently, marketed anti-platelet drugs are associated with major bleeding complications for this exact reason; they are not effective in targeting pathological thrombi without interfering with normal hemostasis. Recent studies have emphasized the importance of shear forces generated from blood flow, that primarily drive platelet activation and aggregation in thrombosis. Local shear stresses in obstructed blood vessels can be higher by up to two orders of magnitude as compared to healthy vessels. Leveraging abnormal shear forces in the thrombus microenvironment may allow to differentiate between thrombosis and hemostasis and develop shear-selective anti-platelet therapies. In this review, we discuss the influence of shear forces on thrombosis and the underlying mechanisms of shear-induced platelet activation. Later, we summarize the therapeutic approaches to target shear-sensitive platelet activation and pathological thrombus growth, with a particular focus on the shear-sensitive protein von Willebrand Factor (VWF). Inhibition of shear-specific platelet aggregation and targeted drug delivery may prove to be much safer and efficacious approaches over current state-of-the-art antithrombotic drugs in the treatment of cardiovascular diseases.
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Affiliation(s)
- Akshita Rana
- Nanobiotechnology Laboratory, Australian Centre for Blood Diseases, Central Clinical School, Monash University, Melbourne, VIC, Australia
| | - Erik Westein
- Nanobiotechnology Laboratory, Australian Centre for Blood Diseases, Central Clinical School, Monash University, Melbourne, VIC, Australia
| | - Be'eri Niego
- Nanobiotechnology Laboratory, Australian Centre for Blood Diseases, Central Clinical School, Monash University, Melbourne, VIC, Australia
| | - Christoph E Hagemeyer
- Nanobiotechnology Laboratory, Australian Centre for Blood Diseases, Central Clinical School, Monash University, Melbourne, VIC, Australia
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23
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Qin RR, Zhu H, Wang F, Song M, Lin PL, Xing YQ, Zhang W, Zhong M, Wang ZH. Platelet activation in diabetic mice models: the role of vascular endothelial cell-derived protein disulfide isomerase-mediated GP IIb/IIIa receptor activation. Aging (Albany NY) 2019; 11:6358-6370. [PMID: 31437127 PMCID: PMC6738422 DOI: 10.18632/aging.102192] [Citation(s) in RCA: 9] [Impact Index Per Article: 1.5] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/11/2019] [Accepted: 08/10/2019] [Indexed: 01/01/2023]
Abstract
GP IIb/IIIa receptor activation plays an important role in thrombosis. The mechanism of early activation of GP IIb/IIIa receptors in diabetic conditions remains unknown. The purpose of this study was to investigate the release of Endothelial microparticle (EMP)-associated protein disulfide isomerase (PDI) after endothelial cell injury induced in diabetes and the changes in platelet activation. We produced an animal model of type 2 diabetes mellitus using ApoE-/- mice. Normal ApoE-/- and diabetic mice were allocated to four groups (n = 15): normal diet, normal diet plus rutin, diabetic, and diabetes plus rutin. The EMP-PDI content and GP IIb/IIIa expression of mice platelets were determined. In addition, EMPs obtained from the four groups were pretreated with the PDI inhibitor rutin; then, their effects on the platelets of normal C57 mice were characterized. Compared with the normal diet group, the diabetic group had significantly increased plasma EMP-PDI content and accelerated platelet activation by increased GP IIb/IIIa expression. In conclusion, EMP-PDI promotes early platelet activation through glycoprotein (GP) IIb/IIIa receptors present on platelet surface in the diabetic state. However, this process could be partially suppressed by the administration of rutin.
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Affiliation(s)
- Ran-Ran Qin
- The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education, Chinese National Health Commission and Chinese Academy of Medical Sciences, The State and Shandong Province Joint Key Laboratory of Translational Cardiovascular Medicine, Department of Cardiology, Qilu Hospital of Shandong University, Jinan, Shandong 250012, China.,Department of Cardiology, The Affiliated Cardiovascular Hospital of Qingdao University, Qingdao, Shandong 266071, China
| | - Hui Zhu
- The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education, Chinese National Health Commission and Chinese Academy of Medical Sciences, The State and Shandong Province Joint Key Laboratory of Translational Cardiovascular Medicine, Department of Cardiology, Qilu Hospital of Shandong University, Jinan, Shandong 250012, China
| | - Feng Wang
- The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education, Chinese National Health Commission and Chinese Academy of Medical Sciences, The State and Shandong Province Joint Key Laboratory of Translational Cardiovascular Medicine, Department of Cardiology, Qilu Hospital of Shandong University, Jinan, Shandong 250012, China
| | - Ming Song
- The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education, Chinese National Health Commission and Chinese Academy of Medical Sciences, The State and Shandong Province Joint Key Laboratory of Translational Cardiovascular Medicine, Department of Cardiology, Qilu Hospital of Shandong University, Jinan, Shandong 250012, China
| | - Pei-Lin Lin
- Department of Cardiology, The Affiliated Cardiovascular Hospital of Qingdao University, Qingdao, Shandong 266071, China
| | - Yan-Qiu Xing
- Department of Geriatric Medicine, Qilu Hospital of Shandong University, Key Laboratory of Cardiovascular Proteomics of Shandong Province, Qilu Hospital of Shandong University, Jinan, Shandong 250012, China
| | - Wei Zhang
- The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education, Chinese National Health Commission and Chinese Academy of Medical Sciences, The State and Shandong Province Joint Key Laboratory of Translational Cardiovascular Medicine, Department of Cardiology, Qilu Hospital of Shandong University, Jinan, Shandong 250012, China
| | - Ming Zhong
- The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education, Chinese National Health Commission and Chinese Academy of Medical Sciences, The State and Shandong Province Joint Key Laboratory of Translational Cardiovascular Medicine, Department of Cardiology, Qilu Hospital of Shandong University, Jinan, Shandong 250012, China
| | - Zhi-Hao Wang
- The Key Laboratory of Cardiovascular Remodeling and Function Research, Chinese Ministry of Education, Chinese National Health Commission and Chinese Academy of Medical Sciences, The State and Shandong Province Joint Key Laboratory of Translational Cardiovascular Medicine, Department of Cardiology, Qilu Hospital of Shandong University, Jinan, Shandong 250012, China.,Department of Geriatric Medicine, Qilu Hospital of Shandong University, Key Laboratory of Cardiovascular Proteomics of Shandong Province, Qilu Hospital of Shandong University, Jinan, Shandong 250012, China
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24
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Daidone V, Galletta E, De Marco L, Casonato A. Cryptic non-canonical splice site activation is part of the mechanism that abolishes multimer organization in the c.2269_2270del von Willebrand factor. Haematologica 2019; 105:1120-1128. [PMID: 31320553 PMCID: PMC7109749 DOI: 10.3324/haematol.2019.222679] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/02/2019] [Accepted: 07/10/2019] [Indexed: 12/20/2022] Open
Abstract
We report a new pathogenic mechanism in von Willebrand disease involving the use of a non-canonical splicing site. The proband, carrying the homozygous c.2269_2270del mutation previously classified as a type 3 mutation, showed severely reduced plasma and platelet von Willebrand factor antigen levels and functions, and no factor VIII binding capacity. A particular von Willebrand factor multimer pattern emerged in plasma, characterized by the presence of only two oligomers: the dimer and an unusually large band, with no intermediate components. There were von Willebrand factor multimers in platelets, but each band ran more slowly than the normal counterpart. No anti-von Willebrand factor antibodies were detectable. The proband was classified as having severe type 1 von Willebrand disease. Seeking the relationship between phenotype and genotype, we found the c.2269_2270del mutation associated with three different RNA: r.2269_2270del (RNAI), giving a truncated von Willebrand factor protein; r.[2269_2270del;2282_2288del] (RNAII), resulting from activation of a cryptic “AG” splicing site; and r.[2269_2270del;2281_2282insAG] (RNAIII), where the wild-type “AG” acceptor of exon 18 was retained due to the non-canonical 2279-2280 “CG” acceptor splicing site being used. The aberrant RNAII and RNAIII caused the alteration of the furin cleavage and binding sites, respectively, both resulting in a von Willebrand factor protein characterized by the persistence of von Willebrand factor propeptide, as confirmed by western blot analysis of the recombinant mutated von Willebrand factor molecules produced in vitro. Taken together, these findings explain the residual von Willebrand factor synthesis, slower-running multimers, and absent factor VIII binding capacity. The apparently pure gene null mutation c.2269_2270del profoundly alters von Willebrand factor gene splicing, inducing a complex RNA expression pattern.
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Affiliation(s)
- Viviana Daidone
- University of Padua Medical School, Depar tment of Medicine, First Chair of Internal Medicine, Padua
| | - Eva Galletta
- University of Padua Medical School, Depar tment of Medicine, First Chair of Internal Medicine, Padua
| | - Luigi De Marco
- IRCCS, C.R.O. Aviano, Depar tment of Translational Research, Stem Cells Unit, Aviano, Italy
| | - Alessandra Casonato
- University of Padua Medical School, Depar tment of Medicine, First Chair of Internal Medicine, Padua
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25
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Jagau H, Behrens IK, Lahme K, Lorz G, Köster RW, Schneppenheim R, Obser T, Brehm MA, König G, Kohler TP, Rohde M, Frank R, Tegge W, Fulde M, Hammerschmidt S, Steinert M, Bergmann S. Von Willebrand Factor Mediates Pneumococcal Aggregation and Adhesion in Blood Flow. Front Microbiol 2019; 10:511. [PMID: 30972039 PMCID: PMC6443961 DOI: 10.3389/fmicb.2019.00511] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.8] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/06/2018] [Accepted: 02/27/2019] [Indexed: 12/24/2022] Open
Abstract
Streptococcus pneumoniae is a major cause of community acquired pneumonia and septicaemia in humans. These diseases are frequently associated with thromboembolic cardiovascular complications. Pneumococci induce the exocytosis of endothelial Weibel-Palade Bodies and thereby actively stimulate the release of von Willebrand factor (VWF), which is an essential glycoprotein of the vascular hemostasis. Both, the pneumococcus induced pulmonary inflammation and the thromboembolytic complications are characterized by a dysbalanced hemostasis including a marked increase in VWF plasma concentrations. Here, we describe for the first time VWF as a novel interaction partner of capsulated and non-encapsulated pneumococci. Moreover, cell culture infection analyses with primary endothelial cells characterized VWF as bridging molecule that mediates bacterial adherence to endothelial cells in a heparin-sensitive manner. Due to the mechanoresponsive changes of the VWF protein conformation and multimerization status, which occur in the blood stream, we used a microfluidic pump system to generate shear flow-induced multimeric VWF strings on endothelial cell surfaces and analyzed attachment of RFP-expressing pneumococci in flow. By applying immunofluorescence visualization and additional electron microscopy, we detected a frequent and enduring bacterial attachment to the VWF strings. Bacterial attachment to the endothelium was confirmed in vivo using a zebrafish infection model, which is described in many reports and acknowledged as suitable model to study hemostasis mechanisms and protein interactions of coagulation factors. Notably, we visualized the recruitment of zebrafish-derived VWF to the surface of pneumococci circulating in the blood stream and detected a VWF-dependent formation of bacterial aggregates within the vasculature of infected zebrafish larvae. Furthermore, we identified the surface-exposed bacterial enolase as pneumococcal VWF binding protein, which interacts with the VWF domain A1 and determined the binding kinetics by surface plasmon resonance. Subsequent epitope mapping using an enolase peptide array indicates that the peptide 181YGAEIFHALKKILKS195 might serve as a possible core sequence of the VWF interaction site. In conclusion, we describe a VWF-mediated mechanism for pneumococcal anchoring within the bloodstream via surface-displayed enolase, which promotes intravascular bacterial aggregation.
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Affiliation(s)
- Hilger Jagau
- Institute of Microbiology, Technische Universität Braunschweig, Braunschweig, Germany
| | - Ina-Kristin Behrens
- Institute of Microbiology, Technische Universität Braunschweig, Braunschweig, Germany
| | - Karen Lahme
- Institute of Microbiology, Technische Universität Braunschweig, Braunschweig, Germany
| | - Georgina Lorz
- Institute of Microbiology, Technische Universität Braunschweig, Braunschweig, Germany
| | - Reinhard W Köster
- Zoological Institute, Technische Universität Braunschweig, Braunschweig, Germany
| | - Reinhard Schneppenheim
- Department of Pediatric Hematology and Oncology, University Medical Center Hamburg-Eppendorf (UKE Hamburg), Hamburg, Germany
| | - Tobias Obser
- Department of Pediatric Hematology and Oncology, University Medical Center Hamburg-Eppendorf (UKE Hamburg), Hamburg, Germany
| | - Maria A Brehm
- Department of Pediatric Hematology and Oncology, University Medical Center Hamburg-Eppendorf (UKE Hamburg), Hamburg, Germany
| | - Gesa König
- Department of Pediatric Hematology and Oncology, University Medical Center Hamburg-Eppendorf (UKE Hamburg), Hamburg, Germany
| | - Thomas P Kohler
- Department of Molecular Genetics and Infection Biology, Interfaculty Institute for Genetics and Functional Genomics, Universität Greifswald, Greifswald, Germany
| | - Manfred Rohde
- Helmholtz Centre for Infection Research, Central Facility for Microscopy, Braunschweig, Germany
| | - Ronald Frank
- Department of Chemical Biology, Helmholtz Centre for Infection Research, Braunschweig, Germany
| | - Werner Tegge
- Department of Chemical Biology, Helmholtz Centre for Infection Research, Braunschweig, Germany
| | - Marcus Fulde
- Centre for Infection Medicine, Institute of Microbiology and Epizootics, Freie Universität Berlin, Berlin, Germany
| | - Sven Hammerschmidt
- Department of Molecular Genetics and Infection Biology, Interfaculty Institute for Genetics and Functional Genomics, Universität Greifswald, Greifswald, Germany
| | - Michael Steinert
- Institute of Microbiology, Technische Universität Braunschweig, Braunschweig, Germany.,Department of Molecular Infection Biology, Helmholtz Centre for Infection Research, Braunschweig, Germany
| | - Simone Bergmann
- Institute of Microbiology, Technische Universität Braunschweig, Braunschweig, Germany
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26
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Von Willebrand factor and the aortic valve: Concepts that are important in the transcatheter aortic valve replacement era. Thromb Res 2018; 170:20-27. [PMID: 30092557 DOI: 10.1016/j.thromres.2018.07.028] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/19/2018] [Revised: 07/17/2018] [Accepted: 07/27/2018] [Indexed: 12/14/2022]
Abstract
Since the approval of the first transcatheter aortic valve replacement (TAVR) device in 2011, this technology has undergone substantial enhancements and exponential growth. However, valve thrombosis and residual paravalvular leaks (PVL) are among the challenges that require further investigation. Recently, monitoring von Willebrand factor (vWF) multimers has emerged as a tool to help evaluate the severity of PVL after TAVR. Following TAVR, vWF large multimers recovery have been documented. The role of large vWF multimers recovery and their interactions with platelets, and the endothelium have not been entirely elucidated. In this review, we discuss vWF synthesis and its role in aortic stenosis. We further provide an overview of the studies that investigated changes affecting vWF multimers following TAVR and the role of HMW vWF multimers monitoring in the determination of PVL severity. We also offer potential future directions for what will be fertile ground for research in this field.
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27
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Rambøl MH, Hisdal J, Sundhagen JO, Brinchmann JE, Rosales A. Recellularization of Decellularized Venous Grafts Using Peripheral Blood: A Critical Evaluation. EBioMedicine 2018; 32:215-222. [PMID: 29779699 PMCID: PMC6020714 DOI: 10.1016/j.ebiom.2018.05.012] [Citation(s) in RCA: 17] [Impact Index Per Article: 2.4] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/13/2018] [Revised: 04/29/2018] [Accepted: 05/08/2018] [Indexed: 01/08/2023] Open
Abstract
Vascular disease is a major cause of death worldwide, and the growing need for replacement vessels is not fully met by autologous grafts or completely synthetic alternatives. Tissue engineering has emerged as a compelling strategy for the creation of blood vessels for reconstructive surgeries. One promising method to obtain a suitable vessel scaffold is decellularization of donor vascular tissue followed by recellularization with autologous cells. To prevent thrombosis of vascular grafts, a confluent and functional autologous endothelium is required, and researchers are still looking for the optimal cell source and recellularization procedure. Recellularization of a decellularized scaffold with only a small volume of whole blood was recently put forward as a feasible option. Here we show that, in contrast to the published results, this method fails to re-endothelialize decellularized veins. Only occasional nucleated cells were seen on the luminal surface of the scaffolds. Instead, we saw fibrin threads, platelets and scattered erythrocytes. Molecular remnants of the endothelial cells were still attached to the scaffold, which explains in part why earlier results were misinterpreted. Decellularized vascular tissues may still be the best scaffolds available for vascular tissue engineering. However, for the establishment of an adequate autologous endothelial lining, methods other than exposure to autologous whole blood need to be developed.
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Affiliation(s)
- Mia H Rambøl
- Norwegian center for stem cell research, Department of immunology, Oslo university hospital, Oslo, Norway; Oslo vascular center, Department of vascular surgery, Oslo university hospital, Oslo, Norway.
| | - Jonny Hisdal
- Oslo vascular center, Department of vascular surgery, Oslo university hospital, Oslo, Norway
| | - Jon O Sundhagen
- Oslo vascular center, Department of vascular surgery, Oslo university hospital, Oslo, Norway
| | - Jan E Brinchmann
- Norwegian center for stem cell research, Department of immunology, Oslo university hospital, Oslo, Norway; Department of molecular medicine, University of Oslo, Oslo, Norway
| | - Antonio Rosales
- Oslo vascular center, Department of vascular surgery, Oslo university hospital, Oslo, Norway
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28
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Makris K, Haliassos A, Chondrogianni M, Tsivgoulis G. Blood biomarkers in ischemic stroke: potential role and challenges in clinical practice and research. Crit Rev Clin Lab Sci 2018; 55:294-328. [DOI: 10.1080/10408363.2018.1461190] [Citation(s) in RCA: 53] [Impact Index Per Article: 7.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/14/2022]
Affiliation(s)
- Konstantinos Makris
- Clinical Biochemistry Department, KAT General Hospital, Kifissia, Athens, Greece
| | | | - Maria Chondrogianni
- Second Department of Neurology, Attikon Hospital, National and Kapodistrian University of Athens, School of Medicine, Athens, Greece
| | - Georgios Tsivgoulis
- Second Department of Neurology, Attikon Hospital, National and Kapodistrian University of Athens, School of Medicine, Athens, Greece
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29
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Gragnano F, Crisci M, Bigazzi MC, Bianchi R, Sperlongano S, Natale F, Fimiani F, Concilio C, Cesaro A, Pariggiano I, Diana V, Limongelli G, Cirillo P, Russo M, Golia E, Calabrò P. Von Willebrand Factor as a Novel Player in Valvular Heart Disease: From Bench to Valve Replacement. Angiology 2018; 69:103-112. [PMID: 28481153 DOI: 10.1177/0003319717708070] [Citation(s) in RCA: 7] [Impact Index Per Article: 1.0] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 12/17/2022]
Abstract
von Willebrand Factor (vWF) is a well-known mediator of hemostasis and vascular inflammation. Its dynamic modulation in the bloodstream, according to hemodynamic conditions, makes it an appealing biomarker in patients with valvular heart disease (VHD). Recent studies highlight the close connection between vWF and VHD, with possible implications in the pathogenesis of VHD, promoting valve aging and calcification or favoring the development of infective endocarditis. Moreover, vWF has been recently proposed as a new diagnostic and prognostic tool in patients with valve stenosis or regurgitation, showing a strict correlation with severity of valve disease, outcome, and bleeding (Heyde syndrome). A novel role for vWF is also emerging in patients undergoing percutaneous or surgical valve repair/replacement to select and stratify patients, evaluate periprocedural bleeding risk, and detect procedural complications. We also report our single-center experience, suggesting, for the first time, possible clinical implications for vWF in percutaneous mitral valve repair (MitraClip). This review summarizes recent advances in the role of vWF in VHD with an updated overview going from bench to operating room.
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Affiliation(s)
- Felice Gragnano
- 1 Division of Cardiology, Department of Cardio-thoracic and Respiratory Sciences, A.O. dei Colli Monaldi Hospital, University of Campania "Luigi Vanvitelli", Naples, Italy
| | - Mario Crisci
- 1 Division of Cardiology, Department of Cardio-thoracic and Respiratory Sciences, A.O. dei Colli Monaldi Hospital, University of Campania "Luigi Vanvitelli", Naples, Italy
| | - Maurizio Cappelli Bigazzi
- 1 Division of Cardiology, Department of Cardio-thoracic and Respiratory Sciences, A.O. dei Colli Monaldi Hospital, University of Campania "Luigi Vanvitelli", Naples, Italy
| | - Renatomaria Bianchi
- 1 Division of Cardiology, Department of Cardio-thoracic and Respiratory Sciences, A.O. dei Colli Monaldi Hospital, University of Campania "Luigi Vanvitelli", Naples, Italy
| | - Simona Sperlongano
- 1 Division of Cardiology, Department of Cardio-thoracic and Respiratory Sciences, A.O. dei Colli Monaldi Hospital, University of Campania "Luigi Vanvitelli", Naples, Italy
| | - Francesco Natale
- 1 Division of Cardiology, Department of Cardio-thoracic and Respiratory Sciences, A.O. dei Colli Monaldi Hospital, University of Campania "Luigi Vanvitelli", Naples, Italy
| | - Fabio Fimiani
- 1 Division of Cardiology, Department of Cardio-thoracic and Respiratory Sciences, A.O. dei Colli Monaldi Hospital, University of Campania "Luigi Vanvitelli", Naples, Italy
| | - Claudia Concilio
- 1 Division of Cardiology, Department of Cardio-thoracic and Respiratory Sciences, A.O. dei Colli Monaldi Hospital, University of Campania "Luigi Vanvitelli", Naples, Italy
| | - Arturo Cesaro
- 1 Division of Cardiology, Department of Cardio-thoracic and Respiratory Sciences, A.O. dei Colli Monaldi Hospital, University of Campania "Luigi Vanvitelli", Naples, Italy
| | - Ivana Pariggiano
- 1 Division of Cardiology, Department of Cardio-thoracic and Respiratory Sciences, A.O. dei Colli Monaldi Hospital, University of Campania "Luigi Vanvitelli", Naples, Italy
| | - Vincenzo Diana
- 1 Division of Cardiology, Department of Cardio-thoracic and Respiratory Sciences, A.O. dei Colli Monaldi Hospital, University of Campania "Luigi Vanvitelli", Naples, Italy
| | - Giuseppe Limongelli
- 1 Division of Cardiology, Department of Cardio-thoracic and Respiratory Sciences, A.O. dei Colli Monaldi Hospital, University of Campania "Luigi Vanvitelli", Naples, Italy
| | - Plinio Cirillo
- 2 Department of Advanced Biological Sciences, Federico II University, Naples, Italy
| | - Mariagiovanna Russo
- 1 Division of Cardiology, Department of Cardio-thoracic and Respiratory Sciences, A.O. dei Colli Monaldi Hospital, University of Campania "Luigi Vanvitelli", Naples, Italy
| | - Enrica Golia
- 1 Division of Cardiology, Department of Cardio-thoracic and Respiratory Sciences, A.O. dei Colli Monaldi Hospital, University of Campania "Luigi Vanvitelli", Naples, Italy
| | - Paolo Calabrò
- 1 Division of Cardiology, Department of Cardio-thoracic and Respiratory Sciences, A.O. dei Colli Monaldi Hospital, University of Campania "Luigi Vanvitelli", Naples, Italy
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Aponte-Santamaría C, Obser T, Grässle S, Oyen F, Budde U, Schneppenheim S, Baldauf C, Gräter F, Schneider SW, Schneppenheim R, Brehm MA, Huck V. von Willebrand disease type 2A phenotypes IIC, IID and IIE: A day in the life of shear-stressed mutant von Willebrand factor. Thromb Haemost 2017; 112:96-108. [DOI: 10.1160/th13-11-0902] [Citation(s) in RCA: 22] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 11/04/2013] [Accepted: 02/11/2014] [Indexed: 11/05/2022]
Abstract
SummaryThe bleeding disorder von Willebrand disease (VWD) is caused by mutations of von Willebrand factor (VWF), a multimeric glycoprotein essential for platelet-dependent primary haemostasis. VWD type 2A–associated mutations each disrupt VWF biosynthesis and function at different stages, depending on the VWF domain altered by the mutation. These effects cause considerable heterogeneity in phenotypes and symptoms. To characterise the molecular mechanisms underlying the specific VWF deficiencies in VWD 2A/IIC, IID and IIE, we investigated VWF variants with patient-derived mutations either in the VWF pro-peptide or in domains D3 or CK. Additionally to static assays and molecular dynamics (MD) simulations we used microfluidic approaches to perform a detailed investigation of the shear-dependent function of VWD 2A mutants. For each group, we found distinct characteristics in their intracellular localisation visualising specific defects in biosynthesis which are correlated to respective multimer patterns. Using microfluidic assays we further determined shear flow-dependent characteristics in polymer-platelet-aggregate formation, platelet binding and string formation for all mutants. The phenotypes observed under flow conditions were not related to the mutated VWF domain. By MD simulations we further investigated how VWD 2A/IID mutations might alter the ability of VWF to form carboxy-terminal dimers. In conclusion, our study offers a comprehensive picture of shear-dependent and shear-independent dysfunction of VWD type 2A mutants. Furthermore, our microfluidic assay might open new possibilities for diagnosis of new VWD phenotypes and treatment choice for VWD patients with shear-dependent VWF dysfunctions that are currently not detectable by static tests.
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de Wee EM, Sanders YV, Mauser-Bunschoten EP, van der Bom JG, Degenaar-Dujardin MEL, Eikenboom J, de Goede-Bolder A, Laros-van Gorkom BAP, Meijer K, Hamulyák K, Nijziel MR, Fijnvandraat K, Leebeek FWG. Determinants of bleeding phenotype in adult patients with moderate or severe von Willebrand disease. Thromb Haemost 2017; 108:683-92. [DOI: 10.1160/th12-04-0244] [Citation(s) in RCA: 67] [Impact Index Per Article: 8.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/17/2012] [Accepted: 07/18/2012] [Indexed: 12/28/2022]
Abstract
SummaryWe performed a nation-wide cross-sectional study to evaluate determinants of bleeding symptoms in a large unselected cohort of adults with von Willebrand disease (VWD). VWD patients were included (n=664), based on lowest historically measured VWF:Ag and VWF:Act levels ≤30 U/dl. Menorrhagia (85%), cutaneous bleeding (77%), bleeding from minor wounds (77%) and oral-cavity bleeding (62%) occurred most frequently. Higher age was associated with a higher bleeding score (BS), determined according to Tosetto, in females. A 10 year increase in age was associated with 0.8 point (95% confidence interval [CI] 0.4–1.1) higher BS. Females had higher BS than males (median 12 vs. 10, p=0.012). BS differed significantly between VWD type 1, 2 and 3: median 9 (-2–31), 13 (-1–33) and 19.5 (1–35), respectively (p<0.001). BS was strongly associated with VWF and FVIII levels: individuals with VWF:Ag levels ≤10 IU/dl, VWF:Act ≤10 IU/dl and FVIII:C ≤10 IU/dl had, respectively, 5.3 point (95%CI 3.2–7.3), 4.3 point (95%CI 2.9–5.8) and 9.6 point (95%CI 6.5–12.7) higher BS, than those with levels >30 IU/dl. In type 3 patients 1 IU/dl FVIII:C decrease was associated with 0.6 point (95% CI 0.1–1.1) BS increase (p=0.021). In conclusion, in VWD patients the bleeding phenotype is strongly associated with type of VWD and VWF and FVIII levels.
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Platelet mitochondrial dysfunction and the correlation with human diseases. Biochem Soc Trans 2017; 45:1213-1223. [PMID: 29054925 DOI: 10.1042/bst20170291] [Citation(s) in RCA: 66] [Impact Index Per Article: 8.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 06/28/2017] [Revised: 09/10/2017] [Accepted: 09/14/2017] [Indexed: 12/20/2022]
Abstract
The platelet is considered as an accessible and valuable tool to study mitochondrial function, owing to its greater content of fully functional mitochondria compared with other metabolically active organelles. Different lines of studies have demonstrated that mitochondria in platelets have function far more than thrombogenesis regulation, and beyond hemostasis, platelet mitochondrial dysfunction has also been used for studying mitochondrial-related diseases. In this review, the interplay between platelet mitochondrial dysfunction and oxidative stress, mitochondrial DNA lesions, electron transfer chain impairments, mitochondrial apoptosis and mitophagy has been outlined. Meanwhile, considerable efforts have been made towards understanding the role of platelet mitochondrial dysfunction in human diseases, such as diabetes mellitus, sepsis and neurodegenerative disorders. Alongside this, we have also articulated our perspectives on the development of potential biomarkers of platelet mitochondrial dysfunction in mitochondrial-related diseases.
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Identification of extant vertebrate Myxine glutinosa VWF: evolutionary conservation of primary hemostasis. Blood 2017; 130:2548-2558. [PMID: 28899852 DOI: 10.1182/blood-2017-02-770792] [Citation(s) in RCA: 11] [Impact Index Per Article: 1.4] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/26/2017] [Accepted: 08/23/2017] [Indexed: 11/20/2022] Open
Abstract
Hemostasis in vertebrates involves both a cellular and a protein component. Previous studies in jawless vertebrates (cyclostomes) suggest that the protein response, which involves thrombin-catalyzed conversion of a soluble plasma protein, fibrinogen, into a polymeric fibrin clot, is conserved in all vertebrates. However, similar data are lacking for the cellular response, which in gnathostomes is regulated by von Willebrand factor (VWF), a glycoprotein that mediates the adhesion of platelets to the subendothelial matrix of injured blood vessels. To gain evolutionary insights into the cellular phase of coagulation, we asked whether a functional vwf gene is present in the Atlantic hagfish, Myxine glutinosa We found a single vwf transcript that encodes a simpler protein compared with higher vertebrates, the most striking difference being the absence of an A3 domain, which otherwise binds collagen under high-flow conditions. Immunohistochemical analyses of hagfish tissues and blood revealed Vwf expression in endothelial cells and thrombocytes. Electron microscopic studies of hagfish tissues demonstrated the presence of Weibel-Palade bodies in the endothelium. Hagfish Vwf formed high-molecular-weight multimers in hagfish plasma and in stably transfected CHO cells. In functional assays, botrocetin promoted VWF-dependent thrombocyte aggregation. A search for vwf sequences in the genome of sea squirts, the closest invertebrate relatives of hagfish, failed to reveal evidence of an intact vwf gene. Together, our findings suggest that VWF evolved in the ancestral vertebrate following the divergence of the urochordates some 500 million years ago and that it acquired increasing complexity though sequential insertion of functional modules.
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Abstract
Beyond its role in hemostasis, von Willebrand factor (VWF) is an emerging mediator of vascular inflammation. Recent studies highlight the involvement of VWF and its regulator, ADAMTS13, in mechanisms that underlie vascular inflammation and immunothrombosis, like leukocyte rolling, adhesion, and extravasation; vascular permeability; ischemia/reperfusion injury; complements activation; and NETosis. The VWF/ADAMTS13 axis is implicated in the pathogenesis of atherosclerosis, promoting plaque formation and inflammation through macrophage and neutrophil recruitment in inflamed lesions. Moreover, VWF and ADAMTS13 have been recently proposed as prognostic biomarkers in cardiovascular, metabolic, and inflammatory diseases, such as diabetes, stroke, myocardial infarction, and sepsis. All these features make VWF an attractive therapeutic target in thromboinflammation. Several lines of research have recently investigated “tailor-made” inhibitors of VWF. Results from animal models and clinical studies support the potent anti-inflammatory and antithrombotic effect of VWF antagonism, providing reassuring data on its safety profile. This review describes the role of VWF in vascular inflammation “from bench to bedside” and provides an updated overview of the drugs that can directly interfere with the VWF/ADAMTS13 axis.
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Abstract
Our study was aimed to investigate anesthetic effects of propofol in patients with different blood groups.A total of 72 participants were enrolled from patients arranged for surgeries of cholecystectomy, tonsillectomy, and spinal operation. Each blood group (A, B, AB, and O) contained 18 participants. Mean arterial pressure (MAP), heart rate (HR), and bispectral index (BIS) were assayed with Philips monitor. These indexes were observed before propofol anesthesia (T0), and then were recorded when concentration of propofol was 1 μg/mL (T1), 2 μg/mL (T2), 3 μg/mL (T3), and 4 μg/mL (T4). The differences in MAP, HR, and BIS at T0 among groups were compared with the χ test. Multiple comparisons were adopted to calculate the differences in MAP, HR, and BIS between groups at T1, T2, T3, and T4.No significant differences in age, sex, and weight of all groups were found (P > .05). Before propofol anesthesia (T0), all the participants exhibited no differences in MAP, HR, and BIS (P > .05). Subsequently, we found obvious differences in ΔMAP, ΔHR, and ΔBIS between groups. The patients in the B blood group showed highest ΔMAP and ΔHR at each time point (P < .05 for both). As for ΔBIS, patients in A blood group exhibited highest value at T3 and T4 (P < .05).The blood group remarkably affects the anesthetic effects of propofol.
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Diagnosis and Treatment of von Willebrand Disease and Rare Bleeding Disorders. J Clin Med 2017; 6:jcm6040045. [PMID: 28394285 PMCID: PMC5406777 DOI: 10.3390/jcm6040045] [Citation(s) in RCA: 39] [Impact Index Per Article: 4.9] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 02/23/2017] [Revised: 04/04/2017] [Accepted: 04/05/2017] [Indexed: 12/19/2022] Open
Abstract
Along with haemophilia A and B, von Willebrand disease (VWD) and rare bleeding disorders (RBDs) cover all inherited bleeding disorders of coagulation. Bleeding tendency, which can range from extremely severe to mild, is the common symptom. VWD, due to a deficiency and/or abnormality of von Willebrand factor (VWF), represents the most frequent bleeding disorder, mostly inherited as an autosomal dominant trait. The diagnosis may be difficult, based on a bleeding history and different diagnostic assays, which evaluate the pleiotropic functions of VWF. Different treatment options are available for optimal management of bleeding and their prevention, and long-term outcomes are generally good. RBDs are autosomal recessive disorders caused by a deficiency of any other clotting factor, apart from factor XII, and cover roughly 5% of all bleeding disorders. The prevalence of the severe forms can range from 1 case in 500,000 up to 1 in 2–3 million, according to the defect. Diagnosis is based on bleeding history, coagulation screening tests and specific factor assays. A crucial problem in RBDs diagnosis is represented by the non-linear relationship between clinical bleeding severity and residual clotting levels; genetic diagnosis may help in understanding the phenotype. Replacement therapies are differently available for patients with RBDs, allowing the successful treatment of the vast majority of bleeding symptoms.
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Lippok S, Radtke M, Obser T, Kleemeier L, Schneppenheim R, Budde U, Netz RR, Rädler JO. Shear-Induced Unfolding and Enzymatic Cleavage of Full-Length VWF Multimers. Biophys J 2017; 110:545-554. [PMID: 26840720 DOI: 10.1016/j.bpj.2015.12.023] [Citation(s) in RCA: 58] [Impact Index Per Article: 7.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/04/2015] [Revised: 11/23/2015] [Accepted: 12/17/2015] [Indexed: 10/22/2022] Open
Abstract
Proteolysis of the multimeric blood coagulation protein von Willebrand Factor (VWF) by ADAMTS13 is crucial for prevention of microvascular thrombosis. ADAMTS13 cleaves VWF within the mechanosensitive A2 domain, which is believed to open under shear flow. In this study, we combine fluorescence correlation spectroscopy (FCS) and a microfluidic shear cell to monitor real-time kinetics of full-length VWF proteolysis as a function of shear stress. For comparison, we also measure the Michaelis-Menten kinetics of ADAMTS13 cleavage of wild-type VWF in the absence of shear but partially denaturing conditions. Under shear, ADAMTS13 activity on full-length VWF arises without denaturing agent as evidenced by FCS and gel-based multimer analysis. In agreement with Brownian hydrodynamics simulations, we find a sigmoidal increase of the enzymatic rate as a function of shear at a threshold shear rate γ˙1/2 = 5522/s. The same flow-rate dependence of ADAMTS13 activity we also observe in blood plasma, which is relevant to predict hemostatic dysfunction.
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Affiliation(s)
- Svenja Lippok
- Faculty of Physics and Center for NanoScience, Ludwig Maximilian University, Munich, Germany
| | - Matthias Radtke
- Department of Physics, Freie Universität Berlin, Berlin, Germany
| | - Tobias Obser
- Department of Pediatric Hematology and Oncology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
| | - Lars Kleemeier
- Faculty of Physics and Center for NanoScience, Ludwig Maximilian University, Munich, Germany
| | - Reinhard Schneppenheim
- Department of Pediatric Hematology and Oncology, University Medical Center Hamburg-Eppendorf, Hamburg, Germany
| | - Ulrich Budde
- Coagulation Lab, Medilys Laborgesellschaft Hamburg, Hamburg, Germany
| | - Roland R Netz
- Department of Physics, Freie Universität Berlin, Berlin, Germany
| | - Joachim O Rädler
- Faculty of Physics and Center for NanoScience, Ludwig Maximilian University, Munich, Germany.
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Yu D, Li S, Wang S, Li X, Zhu M, Huang S, Sun L, Zhang Y, Liu Y, Wang S. Development and Characterization of VEGF165-Chitosan Nanoparticles for the Treatment of Radiation-Induced Skin Injury in Rats. Mar Drugs 2016; 14:md14100182. [PMID: 27727163 PMCID: PMC5082330 DOI: 10.3390/md14100182] [Citation(s) in RCA: 10] [Impact Index Per Article: 1.1] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 08/12/2016] [Revised: 08/30/2016] [Accepted: 08/31/2016] [Indexed: 12/20/2022] Open
Abstract
Radiation-induced skin injury, which remains a serious concern in radiation therapy, is currently believed to be the result of vascular endothelial cell injury and apoptosis. Here, we established a model of acute radiation-induced skin injury and compared the effect of different vascular growth factors on skin healing by observing the changes of microcirculation and cell apoptosis. Vascular endothelial growth factor (VEGF) was more effective at inhibiting apoptosis and preventing injury progression than other factors. A new strategy for improving the bioavailability of vascular growth factors was developed by loading VEGF with chitosan nanoparticles. The VEGF-chitosan nanoparticles showed a protective effect on vascular endothelial cells, improved the local microcirculation, and delayed the development of radioactive skin damage.
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Affiliation(s)
- Daojiang Yu
- Department of Plastic Surgery, the Second Affiliated Hospital, Soochow University, Suzhou 215004, China.
- Department of Pathology, School of Biology & Basic Medical Sciences, Soochow University, Suzhou 215123, China.
| | - Shan Li
- College of Pharmaceutical Science, Soochow University, Suzhou 215123, China.
| | - Shuai Wang
- Department of Plastic Surgery, the Second Affiliated Hospital, Soochow University, Suzhou 215004, China.
| | - Xiujie Li
- Department of Plastic Surgery, the Second Affiliated Hospital, Soochow University, Suzhou 215004, China.
| | - Minsheng Zhu
- Department of Pathology, School of Biology & Basic Medical Sciences, Soochow University, Suzhou 215123, China.
| | - Shai Huang
- Department of Pathology, School of Biology & Basic Medical Sciences, Soochow University, Suzhou 215123, China.
| | - Li Sun
- Department of Pathology, School of Biology & Basic Medical Sciences, Soochow University, Suzhou 215123, China.
| | - Yongsheng Zhang
- Department of Pathology, the Second Affiliated Hospital of Soochow University, Suzhou 215004, China.
| | - Yanli Liu
- College of Pharmaceutical Science, Soochow University, Suzhou 215123, China.
| | - Shouli Wang
- Department of Pathology, School of Biology & Basic Medical Sciences, Soochow University, Suzhou 215123, China.
- Institute of Radiology & Oncology, Soochow University, Suzhou 215006, China.
- Suzhou Key Laboratory of Tumor Microenvironment Pathology, Suzhou 215123, China.
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Michiels JJ, Batorova A, Prigancova T, Smejkal P, Penka M, Vangenechten I, Gadisseur A. Changing insights in the diagnosis and classification of autosomal recessive and dominant von Willebrand diseases 1980-2015. World J Hematol 2016; 5:61-74. [DOI: 10.5315/wjh.v5.i3.61] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Key Words] [Track Full Text] [Download PDF] [Journal Information] [Submit a Manuscript] [Subscribe] [Scholar Register] [Received: 11/26/2015] [Accepted: 04/18/2016] [Indexed: 02/05/2023] Open
Abstract
The European Clinical Laboratory and Molecular (ECLM) criteria define 10 distinct Willebrand diseases (VWD): recessive type 3, severe 1, 2C and 2N; dominant VWD type 1 secretion/clearance defect, 2A, 2B, 2E, 2M and 2D; and mild type 1 VWD (usually carriers of recessive VWD). Recessive severe 1 and 2C VWD are characterized by secretion and multimerization defects caused by mutations in the D1-D2 domain. Recessive 2N VWD is a mild hemophilia due to D’-FVIII-von Willebrand factor (VWF) binding site mutations. Dominant 2E VWD caused by heterozygous missense mutations in the D3 domain is featured by a secretion-clearance-multimerization VWF defect. Dominant VWD type 2M due to loss of function mutations in the A1 domain is characterized by decreased ristocetin-induced platelet aggregation and VWF:RCo, normal VWF multimers and VWF:CB, a poor response of VWF:RCo and good response of VWF:CB to desmopressin (DDAVP). Dominant VWD type 2A induced by heterozygous mutations in the A2 domain results in hypersensitivity of VWF for proteolysis by ADAMTS13 into VWF degradation products, resulting in loss of large VWF multimers with triplet structure of each individual VWF band. Dominant VWD type 2B due to a gain of function mutation in the A1 domain is featured by spontaneous interaction between platelet glycoprotein Ib (GPIb) and mutated VWF A1 followed by increased proteolysis with loss of large VWF multimers and presence of each VWF band. A new category of dominant VWD type 1 secretion or clearance defect due to mutations in the D3 domain or D4-C1-C5 domains consists of two groups: Those with normal or smeary pattern of VWF multimers.
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Li TT, Fan ML, Hou SX, Li XY, Barry DM, Jin H, Luo SY, Kong F, Lau LF, Dai XR, Zhang GH, Zhou LL. A novel snake venom-derived GPIb antagonist, anfibatide, protects mice from acute experimental ischaemic stroke and reperfusion injury. Br J Pharmacol 2015; 172:3904-16. [PMID: 25917571 DOI: 10.1111/bph.13178] [Citation(s) in RCA: 60] [Impact Index Per Article: 6.0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/28/2014] [Revised: 04/02/2015] [Accepted: 04/21/2015] [Indexed: 12/21/2022] Open
Abstract
BACKGROUND AND PURPOSE Ischaemic stroke is a serious disease with limited therapy options. Glycoprotein (GP)Ib binding to von Willebrand factor (vWF) exposed at vascular injury initiates platelet adhesion and contributes to platelet aggregation. GPIb has been suggested as an effective target for antithrombotic therapy in stroke. Anfibatide is a GPIb antagonist derived from snake venom and we investigated its protective effect on experimental brain ischaemia in mice. EXPERIMENTAL APPROACH Focal cerebral ischaemia was induced by 90 min of transient middle cerebral artery occlusion (MCAO). These mice were then treated with anfibatide (4, 2, 1 μg·kg(-1) ), injected i.v., after 90 min of MCAO, followed by 1 h of reperfusion. Tirofiban, a GPIIb/IIIα antagonist, was used as a positive control. KEY RESULTS Twenty-four hours after MCAO, anfibatide-treated mice showed significantly improved ischaemic lesions in a dose-dependent manner. The mice had smaller infarct volumes, less severe neurological deficits and histopathology of cerebrum tissues compared with the untreated MCAO mice. Moreover, anfibatide decreased the amount of GPIbα, vWF and accumulation of fibrin(ogen) in the vasculature of the ischaemic hemisphere. Tirofiban had similar effects on infarct size and fibrin(ogen) deposition compared with the MCAO group. Importantly, the anfibatide-treated mice showed a lower incidence of intracerebral haemorrhage and shorter tail bleeding time compared with the tirofiban-treated mice. CONCLUSIONS AND IMPLICATIONS Our data indicate anfibatide is a safe GPIb antagonist that exerts a protective effect on cerebral ischaemia and reperfusion injury. Anfibatide is a promising candidate that could be beneficial for the treatment of ischaemic stroke.
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Affiliation(s)
- Ting-Ting Li
- Department of Pharmacology, School of Basic Medical Science, Anhui Medical University, Hefei, Anhui, China
| | - Man-Li Fan
- Department of Pharmacology, School of Basic Medical Science, Anhui Medical University, Hefei, Anhui, China
| | - Shi-Xiang Hou
- Department of Pharmacology, School of Basic Medical Science, Anhui Medical University, Hefei, Anhui, China.,Department of Pharmacy, Xuancheng People's Hospital, Xuancheng, China
| | - Xiao-Yi Li
- Zhaoke Pharmaceutical Co. Ltd, Hefei, Anhui, China
| | - Devin M Barry
- Department of Anesthesiology, School of Medicine, Washington University, St. Louis, MO, USA
| | - Hui Jin
- Department of Pharmaceutical Chemistry, Jiangsu Changjiang Pharmaceutical Co. Ltd, Shanghai, China
| | - Sheng-Yong Luo
- Department of Pharmacology, Anhui Academy of Medical Sciences, Hefei, Anhui, China
| | - Feng Kong
- Department of Pharmacology, School of Basic Medical Science, Anhui Medical University, Hefei, Anhui, China
| | - Lit-Fui Lau
- Zhaoke Pharmaceutical Co. Ltd, Hefei, Anhui, China
| | | | - Guo-Hui Zhang
- Department of Pharmacy, Xuancheng People's Hospital, Xuancheng, China.,Zhaoke Pharmaceutical Co. Ltd, Hefei, Anhui, China
| | - Lan-Lan Zhou
- Department of Pharmacology, School of Basic Medical Science, Anhui Medical University, Hefei, Anhui, China
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Successful aortic aneurysm repair in a woman with severe von Willebrand (type 3) disease. Case Rep Hematol 2015; 2015:703803. [PMID: 25960895 PMCID: PMC4417585 DOI: 10.1155/2015/703803] [Citation(s) in RCA: 1] [Impact Index Per Article: 0.1] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 01/19/2015] [Revised: 03/18/2015] [Accepted: 03/19/2015] [Indexed: 11/28/2022] Open
Abstract
von Willebrand disease type 3 (VWD3) is a rare but the most severe form of von Willebrand disease; it is due to almost complete lack of von Willebrand factor activity (VWF:RCo). It is inherited as autosomal recessive trait; whilst heterozygote carriers have mild, or no symptoms, patients with VWD3 show severe bleeding symptoms. In the laboratory, this is characterised by undetectable VWF:Ag, VWF:RCo, and reduced levels of factor VIII < 0.02 IU/dL. The bleeding is managed with von Willebrand/FVIII factor concentrate replacement therapy. In this rare but challenging case we report on the successful excision and repair of an ascending aortic aneurysm following adequate VWF/FVIII factor concentrate replacement using Haemate-P.
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Conventional rapid latex agglutination in estimation of von Willebrand factor: method revisited and potential clinical applications. J Immunol Res 2014; 2014:850810. [PMID: 25759835 PMCID: PMC4352515 DOI: 10.1155/2014/850810] [Citation(s) in RCA: 5] [Impact Index Per Article: 0.5] [Reference Citation Analysis] [Abstract] [Track Full Text] [Download PDF] [Figures] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/04/2014] [Revised: 11/30/2014] [Accepted: 11/30/2014] [Indexed: 12/18/2022] Open
Abstract
Measurement of von Willebrand factor antigen (VWF : Ag) levels is usually performed in a specialised laboratory which limits its application in routine clinical practice. So far, no commercial rapid test kit is available for VWF : Ag estimation. This paper discusses the technical aspect of latex agglutination method which was established to suit the purpose of estimating von Willebrand factor (VWF) levels in the plasma sample. The latex agglutination test can be performed qualitatively and semiquantitatively. Reproducibility, stability, linearity, limit of detection, interference, and method comparison studies were conducted to evaluate the performance of this test. Semiquantitative latex agglutination test was strongly correlated with the reference immunoturbidimetric assay (Spearman's rho = 0.946, P < 0.001, n = 132). A substantial agreement (κ = 0.77) was found between qualitative latex agglutination test and the reference assay. Using the scoring system for the rapid latex test, no agglutination is with 0% VWF : Ag (control negative), 1+ reaction is equivalent to <20% VWF : Ag, and 4+ reaction indicates >150% VWF : Ag (when comparing with immunoturbidimetric assay). The findings from evaluation studies suggest that latex agglutination method is suitable to be used as a rapid test kit for the estimation of VWF : Ag levels in various clinical conditions associated with high levels and low levels of VWF : Ag.
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Shao X, Ren H, Wang YL, Wang F, Hou G, Huang DN. Targeting antitumor effect of rhTNF-α fusion protein mediated by matrix metalloproteinase-2. Oncol Rep 2014; 33:810-8. [PMID: 25421954 DOI: 10.3892/or.2014.3616] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 09/16/2014] [Accepted: 10/30/2014] [Indexed: 11/05/2022] Open
Abstract
The aim of this study was to examine the tumor therapy, targeting effects and side effects of tumor-targeting rhTNF-α fusion protein mediated by matrix metalloproteinase-2 in an animal model in order to provide experimental data for future development of drugs. The median lethal dose (LD50) was obtained from acute toxicity experiments. The A549 lung cancer xenograft model was established, and then randomly divided into the saline, standard substance, and low-, middle- and high-dose fusion protein experiment groups. Each group was administered drugs for 18 days. The length and width of the xenografts were measured every three days, after which the xenograft growth curve was drawn. The mice were sacrificed in each group following treatment and the tumor volume and weight were measured. The targeting, effectiveness and toxicity of the transformed fusion protein, and pathological changes of tumor and organ tissues were examined by hematoxylin and eosin (H&E) staining. Additionally, biochemical markers were used to detect damage of various organs after protein processing. Cell apoptosis and angiogenesis were determined using terminal deoxynucleotidyltransferase-mediated dUTP nick end-labeling (TUNEL) testing and immunohistochemistry, respectively, in different dose groups. Tumor growth was markedly retarded in the high-dose experimental and standard hTNF-α groups with antitumor rates of 85.91 and 72.25%, respectively, as compared with the control group. Furthermore, the tumor tissue showed obvious apoptosis (the apoptotic index was 78.78 and 66.65%, respectively) and pathological changes in the high-dose experimental and standard hTNF-α groups. Tumor angiogenesis in each fusion protein group was inhibited (P<0.01) and the biochemical markers of various organs were greatly reduced in the high-dose experimental group (P<0.05). This finding indicated that slight toxic effects of fusion proteins were evident for the heart, liver and kidney. The reforming fusion protein can therefore target tumor tissues and efficiently kill tumor cells, with few side effects.
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Affiliation(s)
- Xin Shao
- Institute of Biochemistry and Molecular Biology, Guangdong Medical College, Zhanjiang, Guangdong 524023, P.R. China
| | - Hui Ren
- Institute of Biochemistry and Molecular Biology, Guangdong Medical College, Zhanjiang, Guangdong 524023, P.R. China
| | - Yue-Li Wang
- Institute of Biochemistry and Molecular Biology, Guangdong Medical College, Zhanjiang, Guangdong 524023, P.R. China
| | - Fa Wang
- Institute of Biochemistry and Molecular Biology, Guangdong Medical College, Zhanjiang, Guangdong 524023, P.R. China
| | - Gan Hou
- Department of Clinical Biochemistry, Guangdong Medical College, Dongguan, Guangdong 523808, P.R. China
| | - Di-Nan Huang
- Department of Clinical Biochemistry, Guangdong Medical College, Dongguan, Guangdong 523808, P.R. China
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Daidone V, Barbon G, Pontara E, Cattini GM, Gallinaro L, Zampese E, Pizzo P, Casonato A. Loss of cysteine 584 impairs the storage and release, but not the synthesis of von Willebrand factor. Thromb Haemost 2014; 112:1159-66. [PMID: 25230768 DOI: 10.1160/th14-04-0391] [Citation(s) in RCA: 2] [Impact Index Per Article: 0.2] [Reference Citation Analysis] [Abstract] [Key Words] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/29/2014] [Accepted: 07/22/2014] [Indexed: 11/05/2022]
Abstract
Cysteines play a key part in von Willebrand factor (VWF) dimerisation and polymerisation, and their loss may severely affect VWF structure and function. We report on three patients with type 3 von Willebrand disease carrying the new c.1751G>T missense mutation that induces the substitution of cysteine 584 by phenylalanine (C584F), and the deletion of seven nucleotides in exon 7 (c.729_735del), producing a premature stop codon at position 454 (E244Lfs*211). VWF was almost undetectable in the patients' plasma and platelets, while a single, poorly represented, oligomer emerged on plasma VWF multimer analysis. No post-DDAVP increase in VWF and factor VIII was observed. Expressing human recombinant C584F-VWF in HEK293T cells showed that C584F-VWF was synthesised and multimerised but not secreted - apart from the first oligomer, which was slightly represented in the conditioned medium, with a pattern similar to the patients' plasma VWF. The in vitro expression of the E244Lfs*211-VWF revealed a defective synthesis of the mutated VWF, with a behavior typical of loss of function mutations. Cellular trafficking, investigated in HEK293 cells, indicated a normal C584F-VWF content in the endoplasmic reticulum and Golgi apparatus, confirming the synthesis and multimerisation of C584F-VWF. No pseudo-Weibel Palade bodies were demonstrable, however, suggesting that C584F mutation impairs the storage of C584F-VWF. These findings point to cysteine 584 having a role in the release of VWF and its targeting to pseudo-Weibel Palade bodies in vitro, as well as in its storage and release by endothelial cells in vivo.
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Affiliation(s)
| | | | | | | | | | | | | | - A Casonato
- A. Casonato, Via Ospedale Civile 105, 35128 Padova, Italy, Tel.: +39 049 821 7177, Fax: +39 049 657391, E-mail:
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Kwok J, Gue D, Curtis J, Jackson S. Pharmacists have a key role in raising awareness around risks of intranasal desmopressin. J Am Pharm Assoc (2003) 2014; 54:460-2. [PMID: 25216872 DOI: 10.1331/japha.2014.14538] [Citation(s) in RCA: 0] [Impact Index Per Article: 0] [Reference Citation Analysis] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/23/2022]
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Sanders YV, Giezenaar MA, Laros-van Gorkom BAP, Meijer K, van der Bom JG, Cnossen MH, Nijziel MR, Ypma PF, Fijnvandraat K, Eikenboom J, Mauser-Bunschoten EP, Leebeek FWG. von Willebrand disease and aging: an evolving phenotype. J Thromb Haemost 2014; 12:1066-75. [PMID: 24750783 DOI: 10.1111/jth.12586] [Citation(s) in RCA: 81] [Impact Index Per Article: 7.4] [Reference Citation Analysis] [Abstract] [Key Words] [MESH Headings] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 12/18/2013] [Accepted: 04/14/2014] [Indexed: 11/27/2022]
Abstract
BACKGROUND Because the number of elderly von Willebrand disease (VWD) patients is increasing, the pathophysiology of aging in VWD has become increasingly relevant. OBJECTIVES To assess age-related changes in von Willebrand factor (VWF) and factor VIII (FVIII) levels and to compare age-related differences in bleeding phenotype between elderly VWD patients and those < 65 years. We also studied co-morbidity in elderly patients. PATIENTS/METHODS We included VWD patients with VWF levels ≤ 30 U dL(-1) in the nationwide cross-sectional 'Willebrand in the Netherlands' (WiN-) study. Patients reported bleeding episodes and treatment of VWD in the year preceding inclusion and during life. This was compared between VWD patients older (n = 71) and younger (16-64 years, n = 593) than 65 years. In elderly patients, age-related changes in VWF and FVIII levels were studied longitudinally by including all historically measured levels. All medical records were examined for co-morbidity. RESULTS In elderly type 1 patients, a decade age increase was associated with a 3.5 U dL(-1) (95% CI, -0.6 to 7.6) VWF:Ag increase and 7.1 U dL(-1) (95% CI, 0.7 to 13.4) FVIII:C increase. This increase was not observed in elderly type 2 patients. Elderly type 2 patients reported significantly more bleeding symptoms in the year preceding inclusion than younger patients (16/27, 59% vs. 87/221, 39%; P = 0.048), which was not observed in type 1 VWD. CONCLUSIONS von Willebrand factor parameters and bleeding phenotype evolve with increasing age in VWD. VWF and FVIII levels increase with age in type 1 patients with no mitigation in bleeding phenotype. In type 2 patients VWF parameters do not increase with age and in these patients aging is accompanied by increased bleeding.
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Affiliation(s)
- Y V Sanders
- Department of Hematology, Erasmus University Medical Center, Rotterdam, the Netherlands
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Quantitative and qualitative changes of von Willebrand factor and their impact on mortality in patients with end-stage kidney disease. Blood Coagul Fibrinolysis 2014; 24:719-26. [PMID: 23846000 DOI: 10.1097/mbc.0b013e32836261dd] [Citation(s) in RCA: 3] [Impact Index Per Article: 0.3] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/26/2022]
Abstract
von Willebrand factor (vWF) antigen levels are elevated in patients with end-stage kidney disease (ESKD). We determined the quantitative and qualitative abnormalities of vWF and factors influencing vWF proteolysis in participants with ESKD compared with age-matched controls and determined the association between abnormalities in vWF and mortality over 4 years of follow-up. vWF : Ag and von Willebrand factor propeptide (vWFpp) levels, vWF functional activity (vWF :RCo), vWF multimer profiles, ADAMTS-13, thrombospondin 1 (TSP-1), and interleukin 6 (IL-6) were evaluated before and after a single hemodialysis treatment in 55 individuals with vascular disease and an age-matched group of controls (n = 21). vWF : Ag and vWF activity were significantly higher in the ESKD patients and levels increased further following the dialysis procedure. The percentage of high molecular weight multimers (%HMWMs) was significantly elevated in the ESKD patients compared with controls. TSP-1 was lower and IL-6 was higher providing possible explanation for the increase in %HMWM in ESKD. The %HMWM dropped significantly in the postdialysis sample. Mortality at 4 years was significantly associated with vWF : Ag. There are higher plasma vWF : Ag levels and a small increase in HMWMs in the ESKD milieu. The acute drop in the %HMWM of vWF postdialysis appears to be due to shear forces encountered during the dialysis procedure. The contribution of these abnormalities to either a pro-thrombotic and/or pro-bleeding phenotype in this population requires further study.
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Bebar KN, Sinnott V, Brooks MB. Recurrent hemorrhage caused by type 3 von Willebrand disease in a domestic long-haired cat. J Vet Emerg Crit Care (San Antonio) 2014; 24:326-31. [DOI: 10.1111/vec.12185] [Citation(s) in RCA: 8] [Impact Index Per Article: 0.7] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 05/28/2013] [Accepted: 03/21/2014] [Indexed: 11/28/2022]
Affiliation(s)
- Kimberly N. Bebar
- Cape Cod Veterinary Specialists; 11 Bourne Bridge Approach Buzzards Bay MA 02532
| | - Virginia Sinnott
- Cape Cod Veterinary Specialists; 11 Bourne Bridge Approach Buzzards Bay MA 02532
| | - Marjory B. Brooks
- Department of Population Medicine and Diagnostic Sciences; College of Veterinary Medicine; Cornell University; Ithaca NY 14853
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Spontaneous hemarthrosis in combined Glanzmann thrombasthenia and type 2N von Willebrand disease. Blood Coagul Fibrinolysis 2014; 25:401-4. [DOI: 10.1097/mbc.0000000000000067] [Citation(s) in RCA: 7] [Impact Index Per Article: 0.6] [Reference Citation Analysis] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Indexed: 11/27/2022]
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Lippok S, Obser T, Müller JP, Stierle VK, Benoit M, Budde U, Schneppenheim R, Rädler JO. Exponential size distribution of von Willebrand factor. Biophys J 2014; 105:1208-16. [PMID: 24010664 DOI: 10.1016/j.bpj.2013.07.037] [Citation(s) in RCA: 31] [Impact Index Per Article: 2.8] [Reference Citation Analysis] [Abstract] [Track Full Text] [Journal Information] [Subscribe] [Scholar Register] [Received: 04/03/2013] [Revised: 07/03/2013] [Accepted: 07/24/2013] [Indexed: 12/13/2022] Open
Abstract
Von Willebrand Factor (VWF) is a multimeric protein crucial for hemostasis. Under shear flow, it acts as a mechanosensor responding with a size-dependent globule-stretch transition to increasing shear rates. Here, we quantify for the first time, to our knowledge, the size distribution of recombinant VWF and VWF-eGFP using a multilateral approach that involves quantitative gel analysis, fluorescence correlation spectroscopy, and total internal reflection fluorescence microscopy. We find an exponentially decaying size distribution of multimers for recombinant VWF as well as for VWF derived from blood samples in accordance with the notion of a step-growth polymerization process during VWF biosynthesis. The distribution is solely described by the extent of polymerization, which was found to be reduced in the case of the pathologically relevant mutant VWF-IIC. The VWF-specific protease ADAMTS13 systematically shifts the VWF size distribution toward smaller sizes. This dynamic evolution is monitored using fluorescence correlation spectroscopy and compared to a computer simulation of a random cleavage process relating ADAMTS13 concentration to the degree of VWF breakdown. Quantitative assessment of VWF size distribution in terms of an exponential might prove to be useful both as a valuable biophysical characterization and as a possible disease indicator for clinical applications.
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Affiliation(s)
- Svenja Lippok
- Faculty of Physics and Center for NanoScience, Ludwig Maximilian University, Munich, Germany
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