microRNA profile of control vehicle- and coagulation protease factor VIIa-released endothelial extracellular vesicles

Coagulation protease factor VIIa (FVIIa) is shown to induce anti-inflammatory and barrier protective effects via endothelial cell protein C receptor (EPCR)-dependent, protease-activated receptor-1 (PAR1)-mediated cell signaling. FVII-EPCR-PAR1 signaling also induces the release of extracellular vesicles from endothelial cells. To obtain clues on whether microRNA (miR) carried out by FVIIa-released EEVs contribute to anti-inflammatory and barrier protective effects, we analyzed miR expression in control- and FVIIa-released EEVs by deep sequencing. These data revealed that several anti-inflammatory miR expression was higher (more than 2-fold) in FVIIa-released EEVs compared to control EEVs, the most predominant being miR10a-5p. The differential expression of miR10a-5p and several other abundant miRs were validated by qRT-PCR. Subsequent in vitro and in vivo experiments showed that miR10a in FVIIa-released EEVs contribute to anti-inflammatory and barrier protective effects. Overall design: EEVs were isolated from control vehicle- and FVIIa-treated human umbilical vein endothelial cells (24 h treatment). Equal number of EEVs (1x109) were subjected to microRNA analysis by deep-sequencing (UT Southwestern Core Facility)