title: Activated Stat5 trafficking Via Endothelial Cell-derived Extracellular Vesicles Controls IL-3 Pro-angiogenic Paracrine Action.
abstract: 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.
authors.affiliation: Department of Medical Sciences, University of Turin, Italy.
authors.forename: Giusy
authors.initials: G
authors.lastname: Lombardo
authors.affiliation: Department of Medical Sciences, University of Turin, Italy.
authors.forename: Patrizia
authors.initials: P
authors.lastname: Dentelli
authors.affiliation: Department of Medical Sciences, University of Turin, Italy.
authors.forename: Gabriele
authors.initials: G
authors.lastname: Togliatto
authors.affiliation: Department of Medical Sciences, University of Turin, Italy.
authors.forename: Arturo
authors.initials: A
authors.lastname: Rosso
authors.affiliation: Department of Medical Sciences, University of Turin, Italy.
authors.forename: Maddalena
authors.initials: M
authors.lastname: Gili
authors.affiliation: Department of Medical Sciences, University of Turin, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Gallo
authors.affiliation: Department of Medical Sciences, University of Turin, Italy.
authors.forename: Maria Chiara
authors.initials: MC
authors.lastname: Deregibus
authors.affiliation: Department of Medical Sciences, University of Turin, Italy.
authors.forename: Giovanni
authors.initials: G
authors.lastname: Camussi
authors.affiliation: Department of Medical Sciences, University of Turin, Italy.
authors.forename: Maria Felice
authors.initials: MF
authors.lastname: Brizzi
mesh_terms: D002457:Cell Extracts; D019938:Cyclin D1 / Q000378:metabolism; D042783:Endothelial Cells / Q000378:metabolism*; D004789:Enzyme Activation; D048049:Extracellular Signal-Regulated MAP Kinases / Q000378:metabolism; D000067128:Extracellular Vesicles / Q000378:metabolism* / Q000648:ultrastructure; D006801:Humans; D007377:Interleukin-3 / Q000378:metabolism*; D035683:MicroRNAs / Q000378:metabolism; D008954:Models, Biological; D018919:Neovascularization, Physiologic; D019899:Paracrine Communication; D010766:Phosphorylation; D021381:Protein Transport; D050799:STAT5 Transcription Factor / Q000378:metabolism*
chemical_list: D002457:Cell Extracts; D007377:Interleukin-3; D035683:MicroRNAs; D050799:STAT5 Transcription Factor; D019938:Cyclin D1; D048049:Extracellular Signal-Regulated MAP Kinases