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

Madkhali, Yahya; Featherby, Sophie; Collier, Mary; Maraveyas, Anthony; Greenman, John; Ettelaie, Camille

Authors

Yahya Madkhali

Sophie Featherby

Mary Collier

Anthony Maraveyas

Camille Ettelaie



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.

Citation

Madkhali, Y., Featherby, S., Collier, M., Maraveyas, A., Greenman, J., & Ettelaie, C. (2019). The Ratio of Factor VIIa:Tissue Factor Content within Microvesicles Determines the Differential Influence on Endothelial Cells. TH Open, 03(02), e132-e145. https://doi.org/10.1055/s-0039-1688934

Journal Article Type Article
Acceptance Date Apr 10, 2019
Online Publication Date May 15, 2019
Publication Date 2019-04
Deposit Date Jun 13, 2019
Publicly Available Date Jun 14, 2019
Journal TH Open
Print ISSN 2512-9465
Peer Reviewed Peer Reviewed
Volume 03
Issue 02
Pages e132-e145
DOI https://doi.org/10.1055/s-0039-1688934
Public URL https://hull-repository.worktribe.com/output/1799994
Publisher URL https://www.thieme-connect.de/products/ejournals/html/10.1055/s-0039-1688934