Macrovascular thrombosis is driven by tissue factor derived primarily from the blood vessel wall

Abstract Leukocytes and leukocyte-derived microparticles contain low levels of tissue factor (TF) and incorporate into forming thrombi. Although this circulating pool of TF has been proposed to play a key role in thrombosis, its functional significance relative to that of vascular wall TF is poorly defined. We tested the hypothesis that leukocyte-derived TF contributes to thrombus formation in vivo. Compared to wild-type mice, mice with severe TF deficiency (ie, TF–/–, hTF-Tg+, or “low-TF”) demonstrated markedly impaired thrombus formation after carotid artery injury or inferior vena cava ligation. A bone marrow transplantation strategy was used to modulate levels of leukocyte-derived TF. Transplantation of low-TF marrow into wild-type mice did not suppress arterial or venous thrombus formation. Similarly, transplantation of wild-type marrow into low-TF mice did not accelerate thrombosis. In vitro analyses revealed that TF activity in the blood was very low and was markedly exceeded by that present in the vessel wall. Therefore, our results suggest that thrombus formation in the arterial and venous macrovasculature is driven primarily by TF derived from the blood vessel wall as opposed to leukocytes.

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Vascular Gas6 contributes to thrombogenesis and promotes tissue factor up-regulation after vessel injury in mice

Blood ◽

10.1182/blood-2012-05-433730 ◽

2013 ◽

Vol 121 (4) ◽

pp. 692-699 ◽

Cited By ~ 35

Author(s):

Richard S. Robins ◽

Catherine A. Lemarié ◽

Sandrine Laurance ◽

Meghedi N. Aghourian ◽

Jianqiu Wu ◽

...

Keyword(s):

Tissue Factor ◽

Thrombus Formation ◽

Vascular Wall ◽

Specific Gene ◽

Tissue Factor Expression ◽

Vessel Injury ◽

Factor Expression ◽

Platelet Signaling

Abstract Gas6 (growth-arrest specific gene 6) plays a role in thrombus stabilization. Gas6 null (−/−) mice are protected from lethal venous and arterial thromboembolism through platelet signaling defects induced only by 5μM ADP and 10μM of the thromboxane analog, U46619. This subtle platelet defect, despite a dramatic clinical phenotype, raises the possibility that Gas6 from a source other than platelets contributes to thrombus formation. Thus, we hypothesize that Gas6 derived from the vascular wall plays a role in venous thrombus formation. Bone marrow transplantation and platelet depletion/reconstitution experiments generating mice with selective ablations of Gas6 from either the hematopoietic or nonhematopoietic compartments demonstrate an approximately equal contribution by Gas6 from both compartments to thrombus formation. Tissue factor expression was significantly reduced in the vascular wall of Gas6−/− mice compared with WT. In vitro, thrombin-induced tissue factor expression was reduced in Gas6−/− endothelial cells compared with wild-type endothelium. Taken together, these results demonstrate that vascular Gas6 contributes to thrombus formation in vivo and can be explained by the ability of Gas6 to promote tissue factor expression and activity. These findings support the notion that vascular wall-derived Gas6 may play a pathophysiologic role in venous thromboembolism.

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Hematopoietic cell-derived microparticle tissue factor contributes to fibrin formation during thrombus propagation

Blood ◽

10.1182/blood-2004-03-0935 ◽

2004 ◽

Vol 104 (10) ◽

pp. 3190-3197 ◽

Cited By ~ 248

Author(s):

Janet Chou ◽

Nigel Mackman ◽

Glenn Merrill-Skoloff ◽

Brian Pedersen ◽

Barbara C. Furie ◽

...

Keyword(s):

Bone Marrow ◽

Tissue Factor ◽

Vessel Wall ◽

Thrombus Formation ◽

Hematopoietic Cell ◽

Chimeric Mice ◽

Wild Type ◽

Vessel Injury ◽

Living Mouse

Abstract Tissue factor (TF) is expressed on nonvascular cells and cells within the vessel wall and circulates in blood associated with microparticles. Although blood-borne TF accumulates into the developing thrombus during thrombus formation, the contribution of blood-borne TF and vessel wall TF to thrombin generation in vivo following vessel injury is unknown. To determine the source and role of blood-borne microparticle TF, we studied arterial thrombus formation in a living mouse using intravital microscopy. Platelet, TF, and fibrin accumulation in the developing thrombus was compared in wild-type and low TF mice. Compared to wild-type mice, low TF mice formed very small platelet thrombi lacking TF or fibrin. Wild-type and low TF mice received transplants of bone marrow from wild-type and low TF mice. Arterial thrombi in low TF bone marrow/wild-type chimeric mice had decreased size and decreased TF and fibrin levels. Arterial thrombi in wild-type bone marrow/low TF chimeric mice showed decreased platelet thrombus size but normal TF and fibrin levels. This demonstrates that blood-borne TF associated with hematopoietic cell-derived microparticles contributes to thrombus propagation.

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ENDOTHELIAL GAS6 Contributes to Thrombogenesis and Promotes TISSUE FACTOR Upregulation DURING Vessel Injury

Blood ◽

10.1182/blood.v118.21.702.702 ◽

2011 ◽

Vol 118 (21) ◽

pp. 702-702

Author(s):

Richard Robins ◽

Catherine A Lemarie ◽

Jianqiu Wu ◽

Mark Blostein

Keyword(s):

Tissue Factor ◽

Conflicts Of Interest ◽

Thrombus Formation ◽

Vena Cava ◽

Functional Assay ◽

Mrna Induction ◽

Vessel Injury ◽

Platelet Depletion

Abstract Abstract 702 Gas6 is a vitamin K-dependent, secreted protein that amplifies platelet aggregation and secretion in response to platelet agonists. Gas6−/− mice are protected from experimentally induced lethal venous and arterial thromboembolism. This protection has been attributed to defective aggregation in platelets from gas6−/− mice. However, this platelet phenotype was only observed when platelets were challenged by only one agonist, ADP, and only at a concentrations of 5.0μM. This subtle platelet abnormality resulting in a rather dramatic clinical phenotype raise the possibility that gas6 from a source other than platelets contributes to thrombus formation. We hypothesize that gas6 derived from the endothelium plays a role in venous thrombus formation. Gas6−/− mice are protected against venous thrombosis induced by 0.37 M FeCl3 in the inferior vena cava (IVC). Bone marrow transplantation experiments generating mice with selective ablations of gas6 from either the hematopoietic or non-hematopoietic compartments demonstrate an approximately equal contribution by gas6 from both compartments to thrombus formation. Platelet depletion in wild type or gas6−/− mice followed by reconstitution with platelets from either WT or gas6−/− mice confirm that gas6 from compartments other than the platelet contribute to thrombosis development. Furthermore, gas6−/− mice are hyporesponsive to FeCl3 mediated tissue factor induction in venous endothelium, as observed by immunofluorescence staining and later validated by a functional assay. In addition, in vitro, gas6−/− endothelial cells are hyporesponsive to thrombin mediated tissue factor mRNA induction. Taken together, these results suggest that non-hematopoietic gas6, possibly from the endothelium, contributes to thrombus formation in vivo and can be explained by the ability of gas6 to promote endothelial tissue factor induction. These findings support the notion that endothelial gas6 may play a pathophysiologic role in venous thromboembolism. Disclosures: No relevant conflicts of interest to declare.

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Increased tissue factor expression on circulating monocytes in chronic HIV infection: relationship to in vivo coagulation and immune activation

Blood ◽

10.1182/blood-2009-03-210179 ◽

2010 ◽

Vol 115 (2) ◽

pp. 161-167 ◽

Cited By ~ 166

Author(s):

Nicholas T. Funderburg ◽

Elizabeth Mayne ◽

Scott F. Sieg ◽

Robert Asaad ◽

Wei Jiang ◽

...

Keyword(s):

Hiv Infection ◽

Tissue Factor ◽

Immune Activation ◽

Interleukin 6 ◽

Plasma Levels ◽

Thrombus Formation ◽

Hiv Replication ◽

Increased Risk

Abstract HIV infection is associated with an increased risk of thrombosis; and as antiretroviral therapy has increased the lifespan of HIV-infected patients, their risk for cardiovascular events is expected to increase. A large clinical study found recently that all-cause mortality for HIV+ patients was related to plasma levels of interleukin-6 and to D-dimer products of fibrinolysis. We provide evidence that this elevated risk for coagulation may be related to increased proportions of monocytes expressing cell surface tissue factor (TF, thromboplastin) in persons with HIV infection. Monocyte TF expression could be induced in vitro by lipopolysaccharide and flagellin, but not by interleukin-6. Monocyte expression of TF was correlated with HIV levels in plasma, with indices of immune activation, and with plasma levels of soluble CD14, a marker of in vivo lipopolysaccharide exposure. TF levels also correlated with plasma levels of D-dimers, reflective of in vivo clot formation and fibrinolysis. Thus, drivers of immune activation in HIV disease, such as HIV replication, and potentially, microbial translocation, may activate clotting cascades and contribute to thrombus formation and cardiovascular morbidities in HIV infection.

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Accumulation of Tissue Factor into Developing Thrombi In Vivo Is Dependent upon Microparticle P-Selectin Glycoprotein Ligand 1 and Platelet P-Selectin

Journal of Experimental Medicine ◽

10.1084/jem.20021868 ◽

2003 ◽

Vol 197 (11) ◽

pp. 1585-1598 ◽

Cited By ~ 501

Author(s):

Shahrokh Falati ◽

Qingde Liu ◽

Peter Gross ◽

Glenn Merrill-Skoloff ◽

Janet Chou ◽

...

Keyword(s):

Tissue Factor ◽

Blood Coagulation ◽

Thrombus Formation ◽

Recipient Mouse ◽

Wild Type ◽

Activated Platelets ◽

Injury Model ◽

Platelet Thrombus ◽

Flowing Blood

Using a laser-induced endothelial injury model, we examined thrombus formation in the microcirculation of wild-type and genetically altered mice by real-time in vivo microscopy to analyze this complex physiologic process in a system that includes the vessel wall, the presence of flowing blood, and the absence of anticoagulants. We observe P-selectin expression, tissue factor accumulation, and fibrin generation after platelet localization in the developing thrombus in arterioles of wild-type mice. However, mice lacking P-selectin glycoprotein ligand 1 (PSGL-1) or P-selectin, or wild-type mice infused with blocking P-selectin antibodies, developed platelet thrombi containing minimal tissue factor and fibrin. To explore the delivery of tissue factor into a developing thrombus, we identified monocyte-derived microparticles in human platelet–poor plasma that express tissue factor, PSGL-1, and CD14. Fluorescently labeled mouse microparticles infused into a recipient mouse localized within the developing thrombus, indicating that one pathway for the initiation of blood coagulation in vivo involves the accumulation of tissue factor– and PSGL-1–containing microparticles in the platelet thrombus expressing P-selectin. These monocyte-derived microparticles bind to activated platelets in an interaction mediated by platelet P-selectin and microparticle PSGL-1. We propose that PSGL-1 plays a role in blood coagulation in addition to its known role in leukocyte trafficking.

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Abstract 14366: T-lymphocytes May Contribute to Thrombus Formation in Patients With Acute Coronary Syndrome via Expression of Tissue Factor

Circulation ◽

10.1161/circ.132.suppl_3.14366 ◽

2015 ◽

Vol 132 (suppl_3) ◽

Author(s):

Giovanni Cimmino ◽

Giovanni Ciccarelli ◽

Stefano Conte ◽

Grazia Pellegrino ◽

Luigi Insabato ◽

...

Keyword(s):

T Cells ◽

T Lymphocytes ◽

Tissue Factor ◽

Thrombus Formation ◽

Specific Antigen ◽

Activated T Cells ◽

Coronary Syndrome ◽

Stable Cad

Background: Activation of T-cells plays an important role in the pathophysiology of acute coronary syndromes (ACS). We have previously shown that plaques from ACS patients are characterized by a selective oligoclonal expansion of T-cells, indicating a specific, antigen-mediated recruitment of T-cells within the unstable lesions. We have also previously reported that activated T-cells in vitro express functional Tissue Factor (TF) on their surface. At the moment, however it is not known whether expression of TF by T-cells may contribute to thrombus formation in vivo. Methods: Blood was collected from the aorta and the coronary sinus of 13 NSTEMI and 10 stable CAD patients. CD3+ cells were selectively isolated and TF gene expression (real time PCR)and protein levels (western blot) were evaluated. In additional 7 STEMI patients, thrombotic formation material was obtained from the occluded coronary artery by catheter aspiration during primary PCI. TF expression in CD3+ cells was evaluated by immunohistochemistry and confocal microscopy. Results: Transcardiac TF expression in CD3+ cells was significantly higher in NSTEMI patients as compared to CD3+ cells obtained from stable CAD patients. Interestingly, thrombi aspirated from STEMI patients resulted enriched in CD3+cells, which expressed TF on their surface as shown in the figure. Conclusions: Our data demonstrate that in patients with ACS, T-lymphocytes may express surface TF, thus contributing to the process of thrombus formation. This finding may shed new light into the pathophysiologyof ACS.

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Gene-Eluting Stents: Comparison of Adenoviral and Adeno- Associated Viral Gene Delivery to the Blood Vessel Wall In Vivo

Human Gene Therapy ◽

10.1089/hum.2006.17.741 ◽

2006 ◽

Vol 17 (7) ◽

pp. 741-750 ◽

Cited By ~ 37

Author(s):

Faisal Sharif ◽

Sean O. Hynes ◽

Jill McMahon ◽

Ronan Cooney ◽

Siobhan Conroy ◽

...

Keyword(s):

Gene Delivery ◽

Blood Vessel ◽

Vessel Wall ◽

Viral Gene ◽

Blood Vessel Wall ◽

Viral Gene Delivery

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Characterization of acute reversible systemic hypertension in a model of heme protein-induced renal injury

AJP Renal Physiology ◽

10.1152/ajprenal.1999.277.1.f58 ◽

1999 ◽

Vol 277 (1) ◽

pp. F58-F65 ◽

Cited By ~ 2

Author(s):

David H. Warden ◽

Anthony J. Croatt ◽

Zvonimir S. Katusic ◽

Karl A. Nath

Keyword(s):

Nitric Oxide ◽

Mean Arterial Pressure ◽

Arterial Pressure ◽

Sodium Nitroprusside ◽

Vessel Wall ◽

Heme Proteins ◽

Blood Vessel Wall ◽

Vasodilatory Response

In the glycerol model of renal injury we describe an acute rise in systemic arterial pressure which is attended by a reduced vasodilatory response to acetylcholine in vivo; vasodilatory responses to verapamil, however, were not impaired. Neither arginine nor sodium nitroprusside diminished this rise in blood pressure; N ω-nitro-l-arginine methyl ester (l-NAME) elevated basal mean arterial pressure and markedly blunted the rise in mean arterial pressure following the administration of glycerol. Aortic rings from the glycerol-treated rat demonstrate an impaired vasodilatory response to acetylcholine, an effect not repaired by arginine; the vasodilatory responses to nitric oxide donors, sodium nitroprusside and SIN-1, were also impaired; 8-bromo-cGMP, at higher doses, evinced a vasodilatory response comparable to that observed in the control rings. This pattern of responses was not a nonspecific effect of aortic injury, since aortic rings treated with mercuric chloride, a potent oxidant, displayed an impaired vasodilatory response to acetylcholine but not to sodium nitroprusside. We conclude that in the glycerol model of heme protein-induced tissue injury, there is an acute elevation in mean arterial pressure attended by impaired endothelium-dependent vasodilatation in vitro and in vivo. We suggest that the acute scavenging of nitric oxide by heme proteins depletes the blood vessel wall of its endogenous vasodilator and permeation of heme proteins into the blood vessel wall may contribute to such sustained effects as observed in vitro.

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Inhibition of restenosis by tissue factor pathway inhibitor: in vivo and in vitro evidence for suppressed monocyte chemoattraction and reduced gelatinolytic activity

Blood ◽

10.1182/blood-2003-04-1148 ◽

2004 ◽

Vol 103 (5) ◽

pp. 1653-1661 ◽

Cited By ~ 37

Author(s):

Christoph W. Kopp ◽

Thomas Hölzenbein ◽

Sabine Steiner ◽

Rodrig Marculescu ◽

Helga Bergmeister ◽

...

Keyword(s):

Tissue Factor ◽

Tissue Factor Pathway Inhibitor ◽

Early Stage ◽

Thrombus Formation ◽

Matrix Metalloproteinase 2 ◽

Monocyte Chemoattractant Protein 1 ◽

Markers Of Inflammation ◽

Tissue Factor Pathway

AbstractActivation of inflammatory and procoagulant mechanisms is thought to contribute significantly to the initiation of restenosis, a common complication after balloon angioplasty of obstructed arteries. During this process, expression of tissue factor (TF) represents one of the major physiologic triggers of coagulation that results in thrombus formation and the generation of additional signals leading to vascular smooth muscle cell (VSMC) proliferation and migration. In this study, we have investigated the mechanisms by which inhibition of coagulation at an early stage through overexpression of tissue factor pathway inhibitor (TFPI), an endogenous inhibitor of TF, might reduce restenosis. In a rabbit femoral artery model, percutaneous delivery of TFPI using a recombinant adenoviral vector resulted in a significant reduction of the intimamedia ratio 21 days after injury. Investigating several markers of inflammation and coagulation, we found reduced neointimal expression of monocyte chemoattractant protein-1 (MCP-1), lesional monocyte infiltration, and expression of vascular TF, matrix metalloproteinase-2 (MMP-2), and MMP-9. Moreover, overexpression of TFPI suppressed the autocrine release of platelet-derived growth factor BB (PDGF-BB), MCP-1, and MMP-2 in response to factors VIIa and Xa from VSMCs in vitro and inhibited monocyte TF activity. These results suggest that TFPI exerts its action in vivo through not only thrombotic, but also nonthrombotic mechanisms.

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Platelet JNK1 is involved in secretion and thrombus formation

Blood ◽

10.1182/blood-2009-07-233932 ◽

2010 ◽

Vol 115 (20) ◽

pp. 4083-4092 ◽

Cited By ~ 72

Author(s):

Frédéric Adam ◽

Alexandre Kauskot ◽

Paquita Nurden ◽

Eric Sulpice ◽

Marc F. Hoylaerts ◽

...

Keyword(s):

Platelet Aggregation ◽

Thrombus Formation ◽

Blood Perfusion ◽

Collagen Matrix ◽

In Vivo Model ◽

Wild Type ◽

Platelet Secretion

Abstract The role of c-Jun NH2-terminal kinase 1 (JNK1) in hemostasis and thrombosis remains unclear. We show here, with JNK1-deficient (JNK1−/−) mice, that JNK1 plays an important role in platelet biology and thrombus formation. In tail-bleeding assays, JNK1−/− mice exhibited longer bleeding times than wild-type mice (396 ± 39 seconds vs 245 ± 32 seconds). We also carried out in vitro whole-blood perfusion assays on a collagen matrix under arterial shear conditions. Thrombus formation was significantly reduced for JNK1−/− platelets (51%). In an in vivo model of thrombosis induced by photochemical injury to cecum vessels, occlusion times were 4.3 times longer in JNK1−/− arterioles than in wild-type arterioles. Moreover, in vitro studies carried out in platelet aggregation conditions demonstrated that, at low doses of agonists, platelet secretion was impaired in JNK1−/− platelets, leading to altered integrin αIIbβ3 activation and reduced platelet aggregation, via a mechanism involving protein kinase C. JNK1 thus appears to be essential for platelet secretion in vitro, consistent with its role in thrombus growth in vivo. Finally, we showed that ERK2 and another isoform of JNK affect platelet aggregation through 2 pathways, one dependent and another independent of JNK1.

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