Mesenchymal stem cells release exosomes that transfer miRNAs to endothelial cells and promote angiogenesis

// Min Gong 1, 2 , Bin Yu 1 , Jingcai Wang 1 , Yigang Wang 1 , Min Liu 1 , Christian Paul 1 , Ronald W. Millard 1, 3 , De-Sheng Xiao 4 , Muhammad Ashraf 1, 5 and Meifeng Xu 1 1 Department of Pathology and Laboratory Medicine, University of Cincinnati Medical Center, Cincinnati, Ohio, USA 2 Children’s Nutrition Research Centre, Children’s Hospital of Chongqing Medical University, Chongqing, China 3 Department of Pharmacology and Cell Biophysics, University of Cincinnati Medical Center, Cincinnati, Ohio, USA 4 Department of Preventive Medicine, School of Public Health, Guangzhou Medical University, Guangzhou, Guangdong Province, China 5 Department of Pharmacology, University of Illinois at Chicago, Chicago, Illinois, USA Correspondence to: Meifeng Xu, email: [email protected] Keywords: exosomes, miRNA transfer, mesenchymal stem cells, angiogenesis, miR-30b Received: December 19, 2016      Accepted: March 21, 2017      Published: April 01, 2017 ABSTRACT Mesenchymal stem cells (MSCs) have been found to benefit patients with a variety of ischemic diseases via promoting angiogenesis. It is also well established that exosomes secreted from MSCs deliver bioactive molecules, including microRNAs (miRs) to recipient cells. Therefore, we hypothesized that exosomes secreted from MSCs deliver miRs into endothelial cells and mediate angiogenesis. The pro-angiogenic stimulatory capacity of exosomes was investigated using tube-like structure formation and spheroid-based sprouting of human umbilical vein endothelial cells (HUVECs), and in vivo Matrigel plug assay. The secretion of pro-angiogenic miRs (pro-angiomiRs) from MSCs into culture medium and transfer of the miRs to HUVECs were confirmed using real-time quantitative PCR. Supplementation of the exosome secretion blocker GW4869 (10 μM) reduced the pro-angiomiRs in the MSC-derived conditioned medium (CdM MSC ). Addition of exosomes isolated from CdM MSC could directly 1) promote HUVEC tube-like structure formation in vitro ; 2) mobilize endothelial cells into Matrigel plug subcutaneously transplanted into mice; and 3) increase blood flow inside Matrigel plug. Fluorescence tracking showed that the exosomes were internalized rapidly by HUVECs causing an upregulated expression of pro-angiomiRs in HUVECs. Loss-and-gain function of the pro-angiomiRs (e.g., miR-30b) in MSCs significantly altered the pro-angiogenic properties of these MSC-derived exosomes, which could be associated with the regulation of their targets in HUVECs. These results suggest that exosomal transfer of pro-angiogenic miRs plays an important role in MSC mediated angiogenesis and stem cell-to-endothelial cell communication.