Serum-derived extracellular vesicles (EVs) impact on vascular remodeling and prevent muscle damage in acute hind limb ischemia

• Published in: Scientific reports 2017-08, Vol.7 (1), p.8180-14, Article 8180
• Main Authors: Cavallari, Claudia, Ranghino, Andrea, Tapparo, Marta, Cedrino, Massimo, Figliolini, Federico, Grange, Cristina, Giannachi, Valentina, Garneri, Paolo, Deregibus, Maria Chiara, Collino, Federica, Rispoli, Pietro, Camussi, Giovanni, Brizzi, Maria Felice
• Format: Article
• Language: English
• Subjects: 13; 13/1; 13/105; 13/95; 45/29; 45/61; 45/90; 692/308/2778; 692/699/75/593/1920; Angiogenesis; Biological activity; Chromatin; Deoxyribonucleic acid; DNA; Extracellular matrix; Extracellular vesicles; Growth factors; Humanities and Social Sciences; Immunoprecipitation; Insulin; Ischemia; Matrix metalloproteinase; Metalloproteinase; multidisciplinary; Proteomics; Rodents; Science; Science (multidisciplinary); Signal transduction; Smad protein; Therapeutic applications; Transforming growth factor-b1; Vascular endothelial growth factor
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-017-08250-0

Serum is an abundant and accessible source of circulating extracellular vesicles (EVs). Serum-EV (sEV) pro-angiogenic capability and mechanisms are herein analyzed using an in vitro assay which predicts sEV angiogenic potential in vivo . Effective sEVs (e-sEVs) also improved vascular remodeling and prevented muscle damage in a mouse model of acute hind limb ischemia. e-sEV angiogenic proteomic and transcriptomic analyses show a positive correlation with matrix-metalloproteinase activation and extracellular matrix organization, cytokine and chemokine signaling pathways, Insulin-like Growth Factor and platelet pathways, and Vascular Endothelial Growth Factor signaling. A discrete gene signature, which highlights differences in e-sEV and ineffective-EV biological activity, was identified using gene ontology (GO) functional analysis. An enrichment of genes associated with the Transforming Growth Factor beta 1 (TGFβ1) signaling cascade is associated with e-sEV administration but not with ineffective-EVs. Chromatin immunoprecipitation analysis on the inhibitor of DNA binding I (ID1) promoter region, and the knock-down of small mother against decapentaplegic (SMAD)1–5 proteins confirmed GO functional analyses. This study demonstrates sEV pro-angiogenic activity, validates a simple, sEV pro-angiogenic assay which predicts their biological activity in vivo , and identifies the TGFβ1 cascade as a relevant mediator. We propose serum as a readily available source of EVs for therapeutic purposes.