CC BY 4.0 · TH Open 2019; 03(02): e132-e145
DOI: 10.1055/s-0039-1688934
Original Article
Georg Thieme Verlag KG Stuttgart · New York

The Ratio of Factor VIIa:Tissue Factor Content within Microvesicles Determines the Differential Influence on Endothelial Cells

Yahya Madkhali
1   Department of Biomedical Sciences, University of Hull, Hull, United Kingdom
2   Department of Medical Laboratories, College of Applied Medical Sciences, Majmaah University, KSA, Al Majmaah, Saudi Arabia
,
Sophie Featherby
1   Department of Biomedical Sciences, University of Hull, Hull, United Kingdom
,
Mary E. Collier
3   Department of Cardiovascular Sciences, University of Leicester, Glenfield General Hospital, Leicester, United Kingdom
,
Anthony Maraveyas
4   Division of Cancer–Hull York Medical School, University of Hull, Hull, United Kingdom
,
John Greenman
1   Department of Biomedical Sciences, University of Hull, Hull, United Kingdom
,
1   Department of Biomedical Sciences, University of Hull, Hull, United Kingdom
› Author Affiliations
Further Information

Publication History

31 December 2018

10 April 2019

Publication Date:
15 May 2019 (online)

Abstract

Tissue factor (TF)-positive microvesicles from various sources can promote cellular proliferation or alternatively induce apoptosis, but the determining factors are unknown. In this study the hypothesis that the ratio of fVIIa:TF within microvesicles determines this outcome was examined. Microvesicles were isolated from HepG2, BxPC-3, 786-O, MDA-MB-231, and MCF-7 cell lines and microvesicle-associated fVIIa and TF antigen and activity levels were measured. Human coronary artery endothelial cells (HCAECs) were incubated with these purified microvesicles, or with combinations of fVIIa-recombinant TF, and cell proliferation/apoptosis was measured. Additionally, by expressing mCherry-PAR2 on HCAEC surface, PAR2 activation was quantified. Finally, the activation of PAR2 on HCAEC or the activities of TF and fVIIa in microvesicles were blocked prior to addition of microvesicles to cells. The purified microvesicles exhibited a range of fVIIa:TF ratios with HepG2 and 786-O cells having the highest (54:1) and lowest (10:1) ratios, respectively. The reversal from proapoptotic to proliferative was estimated to occur at a fVIIa:TF molar ratio of 15:1, but HCAEC could not be rescued at higher TF concentrations. The purified microvesicles induced HCAEC proliferation or apoptosis according to this ruling. Blocking PAR2 activation on HCAEC, or inhibiting fVIIa or TF-procoagulant function on microvesicles prevented the influence on HCAEC. Finally, incubation of HCAEC with recombinant TF resulted in increased surface exposure of fVII. The induction of cell proliferation or apoptosis by TF-positive microvesicles is dependent on the ratio of fVIIa:TF and involves the activation of PAR2. At lower TF concentrations, fVIIa can counteract the proapoptotic stimulus and induce proliferation.

Authors' Contributions

The study was designed by Y.M., M.E.C., A.M., and C.E., and the experimental work was performed by Y.M., S.F., and C.E. The data were evaluated by Y.M., M.E.C., J.G., A.M., and C.E. and the manuscript was prepared by Y.M., J.G., and C.E.


Supplementary Material

 
  • References

  • 1 Kirchhofer D, Nemerson Y. Initiation of blood coagulation: the tissue factor/factor VIIa complex. Curr Opin Biotechnol 1996; 7 (04) 386-391
  • 2 Edgington TS, Dickinson CD, Ruf W. The structural basis of function of the TF. VIIa complex in the cellular initiation of coagulation. Thromb Haemost 1997; 78 (01) 401-405
  • 3 Morel O, Toti F, Hugel B. , et al. Procoagulant microparticles: disrupting the vascular homeostasis equation?. Arterioscler Thromb Vasc Biol 2006; 26 (12) 2594-2604
  • 4 Freyssinet JM, Toti F. Formation of procoagulant microparticles and properties. Thromb Res 2010; 125 (Suppl. 01) S46-S48
  • 5 Gardiner C, Harrison P, Belting M. , et al. Extracellular vesicles, tissue factor, cancer and thrombosis - discussion themes of the ISEV 2014 Educational Day. J Extracell Vesicles 2015; 4: 26901
  • 6 Date K, Hall J, Greenman J, Maraveyas A, Madden LA. Tumour and microparticle tissue factor expression and cancer thrombosis. Thromb Res 2013; 131 (02) 109-115
  • 7 Hron G, Kollars M, Weber H. , et al. Tissue factor-positive microparticles: cellular origin and association with coagulation activation in patients with colorectal cancer. Thromb Haemost 2007; 97 (01) 119-123
  • 8 Thaler J, Koder S, Kornek G, Pabinger I, Ay C. Microparticle-associated tissue factor activity in patients with metastatic pancreatic cancer and its effect on fibrin clot formation. Transl Res 2014; 163 (02) 145-150
  • 9 Auwerda JJ, Yuana Y, Osanto S. , et al. Microparticle-associated tissue factor activity and venous thrombosis in multiple myeloma. Thromb Haemost 2011; 105 (01) 14-20
  • 10 Kask L, Jorsback A, Winkvist M. , et al. Identification of novel downstream molecules of tissue factor activation by comparative proteomic analysis. J Proteome Res 2014; 13 (02) 477-488
  • 11 Ramchandani D, Unruh D, Lewis CS, Bogdanov VY, Weber GF. Activation of carbonic anhydrase IX by alternatively spliced tissue factor under late-stage tumor conditions. Lab Invest 2016; 96 (12) 1234-1245
  • 12 Zelaya H, Rothmeier AS, Ruf W. Tissue factor at the crossroad of coagulation and cell signaling. J Thromb Haemost 2018; 16 (10) 1941-1952
  • 13 McVey JH. The role of the tissue factor pathway in haemostasis and beyond. Curr Opin Hematol 2016; 23 (05) 453-461
  • 14 Rothmeier AS, Liu E, Chakrabarty S. , et al. Identification of the integrin-binding site on coagulation factor VIIa required for proangiogenic PAR2 signaling. Blood 2018; 131 (06) 674-685
  • 15 Fan L, Yotov WV, Zhu T. , et al. Tissue factor enhances protease-activated receptor-2-mediated factor VIIa cell proliferative properties. J Thromb Haemost 2005; 3 (05) 1056-1063
  • 16 Cirillo P, Calì G, Golino P. , et al. Tissue factor binding of activated factor VII triggers smooth muscle cell proliferation via extracellular signal-regulated kinase activation. Circulation 2004; 109 (23) 2911-2916
  • 17 Wu B, Zhou H, Hu L, Mu Y, Wu Y. Involvement of PKCα activation in TF/VIIa/PAR2-induced proliferation, migration, and survival of colon cancer cell SW620. Tumour Biol 2013; 34 (02) 837-846
  • 18 Gessler F, Voss V, Dützmann S, Seifert V, Gerlach R, Kögel D. Inhibition of tissue factor/protease-activated receptor-2 signaling limits proliferation, migration and invasion of malignant glioma cells. Neuroscience 2010; 165 (04) 1312-1322
  • 19 Versteeg HH, Ruf W. Emerging insights in tissue factor-dependent signaling events. Semin Thromb Hemost 2006; 32 (01) 24-32
  • 20 Kocatürk B, Van den Berg YW, Tieken C. , et al. Alternatively spliced tissue factor promotes breast cancer growth in a β1 integrin-dependent manner. Proc Natl Acad Sci U S A 2013; 110 (28) 11517-11522
  • 21 Collier ME, Ettelaie C. Induction of endothelial cell proliferation by recombinant and microparticle-tissue factor involves beta1-integrin and extracellular signal regulated kinase activation. Arterioscler Thromb Vasc Biol 2010; 30 (09) 1810-1817
  • 22 Rak J, Milsom C, Yu J. Tissue factor in cancer. Curr Opin Hematol 2008; 15 (05) 522-528
  • 23 Schaffner F, Ruf W. Tissue factor and PAR2 signaling in the tumor microenvironment. Arterioscler Thromb Vasc Biol 2009; 29 (12) 1999-2004
  • 24 Lee BJ, Kim JH, Woo SH, Kim JH, Kim DH, Yu YS. Tissue factor is involved in retinoblastoma cell proliferation via both the Akt and extracellular signal-regulated kinase pathways. Oncol Rep 2011; 26 (03) 665-670
  • 25 Pradier A, Ettelaie C. The influence of exogenous tissue factor on the regulators of proliferation and apoptosis in endothelial cells. J Vasc Res 2008; 45 (01) 19-32
  • 26 Alkistis Frentzou G, Collier ME, Seymour AM, Ettelaie C. Differential induction of cellular proliferation, hypertrophy and apoptosis in H9c2 cardiomyocytes by exogenous tissue factor. Mol Cell Biochem 2010; 345 (1–2): 119-130
  • 27 ElKeeb AM, Collier ME, Maraveyas A, Ettelaie C. Accumulation of tissue factor in endothelial cells induces cell apoptosis, mediated through p38 and p53 activation. Thromb Haemost 2015; 114 (02) 364-378
  • 28 Aharon A, Tamari T, Brenner B. Monocyte-derived microparticles and exosomes induce procoagulant and apoptotic effects on endothelial cells. Thromb Haemost 2008; 100 (05) 878-885
  • 29 Shinagawa K, Ploplis VA, Castellino FJ. A severe deficiency of coagulation factor VIIa results in attenuation of the asthmatic response in mice. Am J Physiol Lung Cell Mol Physiol 2009; 296 (05) L763-L770
  • 30 Camerer E, Huang W, Coughlin SR. Tissue factor- and factor X-dependent activation of protease-activated receptor 2 by factor VIIa. Proc Natl Acad Sci U S A 2000; 97 (10) 5255-5260
  • 31 Riewald M, Ruf W. Mechanistic coupling of protease signaling and initiation of coagulation by tissue factor. Proc Natl Acad Sci U S A 2001; 98 (14) 7742-7747
  • 32 Schaffner F, Ruf W. Tissue factor and protease-activated receptor signaling in cancer. Semin Thromb Hemost 2008; 34 (02) 147-153
  • 33 Hjortoe GM, Petersen LC, Albrektsen T. , et al. Tissue factor-factor VIIa-specific up-regulation of IL-8 expression in MDA-MB-231 cells is mediated by PAR-2 and results in increased cell migration. Blood 2004; 103 (08) 3029-3037
  • 34 Boulanger CM, Scoazec A, Ebrahimian T. , et al. Circulating microparticles from patients with myocardial infarction cause endothelial dysfunction. Circulation 2001; 104 (22) 2649-2652
  • 35 Morel O, Toti F, Hugel B, Freyssinet JM. Cellular microparticles: a disseminated storage pool of bioactive vascular effectors. Curr Opin Hematol 2004; 11 (03) 156-164
  • 36 Morel O, Toti F, Bakouboula B, Grunebaum L, Freyssinet JM. Procoagulant microparticles: ‘criminal partners’ in atherothrombosis and deleterious cellular exchanges. Pathophysiol Haemost Thromb 2006; 35 (1–2): 15-22
  • 37 Morel O, Morel N, Freyssinet JM, Toti F. Platelet microparticles and vascular cells interactions: a checkpoint between the haemostatic and thrombotic responses. Platelets 2008; 19 (01) 9-23
  • 38 Collier ME, Mah PM, Xiao Y, Maraveyas A, Ettelaie C. Microparticle-associated tissue factor is recycled by endothelial cells resulting in enhanced surface tissue factor activity. Thromb Haemost 2013; 110 (05) 966-976
  • 39 Yang Q, Underwood MJ, Hsin MK, Liu XC, He GW. Dysfunction of pulmonary vascular endothelium in chronic obstructive pulmonary disease: basic considerations for future drug development. Curr Drug Metab 2008; 9 (07) 661-667
  • 40 Kuge Y, Kume N, Ishino S. , et al. Prominent lectin-like oxidized low density lipoprotein (LDL) receptor-1 (LOX-1) expression in atherosclerotic lesions is associated with tissue factor expression and apoptosis in hypercholesterolemic rabbits. Biol Pharm Bull 2008; 31 (08) 1475-1482
  • 41 Ettelaie C, Collier MEW, Maraveyas A, Ettelaie R. Characterization of physical properties of tissue factor-containing microvesicles and a comparison of ultracentrifuge-based recovery procedures. J Extracell Vesicles 2014; 3: 10
  • 42 Ettelaie C, Collier MEW, Featherby S, Greenman J, Maraveyas A. Peptidyl-prolyl isomerase 1 (Pin1) preserves the phosphorylation state of tissue factor and prolongs its release within microvesicles. Biochim Biophys Acta Mol Cell Res 2018; 1865 (01) 12-24
  • 43 Benelhaj NE, Maraveyas A, Featherby S, Collier MEW, Johnson MJ, Ettelaie C. Alteration in endothelial permeability occurs in response to the activation of PAR2 by factor Xa but not directly by the TF-factor VIIa complex. Thromb Res 2019; 175: 13-20
  • 44 Collier MEW, Ettelaie C, Goult BT, Maraveyas A, Goodall AH. Investigation of the filamin A-dependent mechanisms of tissue factor incorporation into microvesicles. Thromb Haemost 2017; 117 (11) 2034-2044
  • 45 Ettelaie C, Collier ME, Featherby S, Benelhaj NE, Greenman J, Maraveyas A. Analysis of the potential of cancer cell lines to release tissue factor-containing microvesicles: correlation with tissue factor and PAR2 expression. Thromb J 2016; 14: 2
  • 46 Broze Jr GJ, Hickman S, Miletich JP. Monoclonal anti-human factor VII antibodies. Detection in plasma of a second protein antigenically and genetically related to factor VII. J Clin Invest 1985; 76 (03) 937-946
  • 47 Böhm E, Seyfried BK, Dockal M. , et al. Differences in N-glycosylation of recombinant human coagulation factor VII derived from BHK, CHO, and HEK293 cells. BMC Biotechnol 2015; 15: 87
  • 48 Buendía P, Montes de Oca A, Madueño JA. , et al. Endothelial microparticles mediate inflammation-induced vascular calcification. FASEB J 2015; 29 (01) 173-181
  • 49 Harter PN, Dützmann S, Drott U. , et al. Anti-tissue factor (TF9-10H10) treatment reduces tumor cell invasiveness in a novel migratory glioma model. Neuropathology 2013; 33 (05) 515-525
  • 50 Versteeg HH, Schaffner F, Kerver M. , et al. Inhibition of tissue factor signaling suppresses tumor growth. Blood 2008; 111 (01) 190-199
  • 51 Teplyakov A, Obmolova G, Malia TJ. , et al. Crystal structure of tissue factor in complex with antibody 10H10 reveals the signaling epitope. Cell Signal 2017; 36: 139-144
  • 52 Basavaraj MG, Olsen JO, Østerud B, Hansen JB. Differential ability of tissue factor antibody clones on detection of tissue factor in blood cells and microparticles. Thromb Res 2012; 130 (03) 538-546
  • 53 Liu R, Tan YZ, Wang HJ, Zhang M. Sorting of lymphatic endothelial progenitor cells from canine peripheral blood and their differentiation induction towards endothelial cells [in Chinese]. Zhonghua Xue Ye Xue Za Zhi 2007; 28 (03) 169-173