Serum-derived extracellular vesicles (EVs) impact on vascular remodeling and prevent muscle damage in acute hind limb ischemia.
Serum-derived extracellular vesicles (EVs) impact on vascular remodeling and prevent muscle damage in acute hind limb ischemia.
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DOI：
10.1038/s41598-017-08250-0
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发表时间：
2017-08-15
影响因子：
4.6
通讯作者：
Brizzi MF
中科院分区：
文献类型：
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作者：
Cavallari C;Ranghino A;Tapparo M;Cedrino M;Figliolini F;Grange C;Giannachi V;Garneri P;Deregibus MC;Collino F;Rispoli P;Camussi G;Brizzi MF
血清是循环细胞外囊泡（EV）的丰富且可获得的来源。本文使用预测sEV体内血管生成潜力的体外测定来分析血清EV（sEV）促血管生成能力和机制。有效的sEV（e-sEV）还改善了急性后肢缺血小鼠模型中的血管重塑并预防了肌肉损伤。e-sEV血管生成蛋白质组学和转录组学分析显示与基质金属蛋白酶活化和细胞外基质组织、细胞因子和趋化因子信号传导途径、胰岛素样生长因子和血小板途径以及血管内皮生长因子信号传导正相关。使用基因本体（GO）功能分析鉴定了突出e-sEV和无效EV生物活性差异的离散基因签名。与转化生长因子β 1（TGFβ1）信号级联相关的基因富集与e-sEV给药相关，但与无效EV无关。DNA结合抑制因子I（ID 1）启动子区的染色质免疫沉淀分析和小母细胞对十肢麻痹（SMAD）1-5蛋白的敲低证实了GO功能分析。该研究证明了sEV的促血管生成活性，验证了一种简单的sEV促血管生成测定法，该测定法可预测其体内生物活性，并将TGFβ1级联识别为相关介体。我们提出血清作为用于治疗目的的EV的容易获得的来源。
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.