Activated Stat5 trafficking Via Endothelial Cell-derived Extracellular Vesicles Controls IL-3 Pro-angiogenic Paracrine Action

• Published in: Scientific reports Vol. 6; no. 1; p. 25689
• Main Authors: Lombardo, Giusy, Dentelli, Patrizia, Togliatto, Gabriele, Rosso, Arturo, Gili, Maddalena, Gallo, Sara, Deregibus, Maria Chiara, Camussi, Giovanni, Brizzi, Maria Felice
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 09-05-2016; Nature Publishing Group
• Subjects: 13; 13/1; 13/109; 13/21; 13/31; 13/95; 38/15; 631/1647; 631/80; Angiogenesis; Cell fate; Chromatin; Cyclin D1; Endothelial cells; Extracellular vesicles; Humanities and Social Sciences; Immunoprecipitation; Interleukin 3; multidisciplinary; Paracrine signalling; Science; Signal transduction; Stat5 protein; Transcription; Wound healing
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/srep25689

Soluble factors and cell-derived extracellular vesicles (EVs) control vascular cell fate during inflammation. The present study investigates the impact of Interleukin 3 (IL-3) on EV release by endothelial cells (ECs), the mechanisms involved in EV release and paracrine actions. We found that IL-3 increases EV release, which is prevented by IL-3Ralpha blockade. EVs released upon IL-3 stimulation were able to induce pro-angiogenic signals as shown by chromatin immunoprecipitation (ChIP) assay performed on the promoter region of cyclin D1 and tridimensional tube-like structure formation. We herein demonstrate that these effects rely on the transfer of miR-126-3p, pre-miR-126 and, more importantly, of activated signal transduction and activator of transcription 5 (pSTAT5) from IL-3-EV cargo into recipient ECs. We show, using the dominant negative form (ΔN)STAT5 and an activated STAT5 (1*6STAT5) constructs, that STAT5 drives IL-3-mediated EV release, miR-126-3p and pSTAT5 content. Finally, using EVs recovered from ΔNSTAT5 expressing ECs, we provide evidence that miR-126-3p and pSTAT5 trafficking is relevant for IL-3-mediated paracrine pro-angiogenic signals. These results indicate that IL-3 regulates EC-EV release, cargo and IL-3 angiogenic paracrine action via STAT5. Moreover, these results provide evidence that EC-derived IL-3-EVs can serve as pro-angiogenic clinical delivery wound healing devices.