4.6 Article

A novel pathway of cellular activation mediated by antiphospholipid antibody-induced extracellular vesicles

Journal

JOURNAL OF THROMBOSIS AND HAEMOSTASIS
Volume 13, Issue 10, Pages 1928-1940

Publisher

WILEY
DOI: 10.1111/jth.13072

Keywords

antiphospholipid antibodies; cell-derived microparticles; endothelial cells; exosomes; interleukins; Toll-like receptors

Funding

  1. NHLBI NIH HHS [P50 HL081011, R01 HL123098] Funding Source: Medline

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BackgroundElevated levels of endothelial cell (EC)-derived extracellular vesicles (EVs) circulate in patients with antiphospholipid antibodies (APLAs), and APLAs, particularly those against (2)-glycoproteinI ((2)GPI), stimulate EV release from ECs. However, the effects of EC-derived EVs have not been characterized. ObjectiveTo determine the mechanism by which EVs released from ECs by anti-(2)GPI antibodies activate unstimulated ECs. Patients/methodsWe used interleukin (IL)-1 receptor inhibitors, small interfering RNA (siRNA) against Toll-like receptors (TLRs) and microRNA (miRNA) profiling to assess the mechanism(s) by which EVs released from ECs exposed to anti-(2)GPI antibodies activated unstimulated ECs. Results and conclusionsAnti-(2)GPI antibodies caused formation of an EC inflammasome and the release of EVs that were enriched in mature IL-1, had a distinct miRNA profile, and caused endothelial activation. However, activation was not inhibited by an IL-1 antibody, an IL-1 receptor antagonist, or IL-1 receptor siRNA. EC activation by EVs required IL-1 receptor-associated kinase4 phosphorylation, and was inhibited by pretreatment of cells with TLR7 siRNA or RNaseA, which degrades ssRNA. Profiling of miRNA in EVs released from ECs incubated with (2)GPI and either control IgG or anti-(2)GPI antibodies revealed numerous differences in the content of specific miRNAs, including a significant decrease in mIR126. These observations demonstrate that, although anti-(2)GPI-derived endothelial EVs contain IL-1, they activate unstimulated ECs through a TLR7-dependent and ssRNA-dependent pathway. Alterations in miRNA content may contribute to the ability of EVs derived from ECs exposed to anti-(2)GPI antibodies to activate unstimulated ECs in an autocrine or paracrine manner.

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