title: MicroRNA-377 regulates mesenchymal stem cell-induced angiogenesis in ischemic hearts by targeting VEGF.
abstract: MicroRNAs have been appreciated in various cellular functions, including the regulation of angiogenesis. Mesenchymal-stem-cells (MSCs) transplanted to the MI heart improve cardiac function through paracrine-mediated angiogenesis. However, whether microRNAs regulate MSC induced angiogenesis remains to be clarified. Using microRNA microarray analysis, we identified a microRNA expression profile in hypoxia-treated MSCs and observed that among all dysregulated microRNAs, microRNA-377 was decreased the most significantly. We also validated that vascular endothelial growth factor (VEGF) is a target of microRNA-377 using dual-luciferase reporter assay and Western-blotting. Knockdown of endogenous microRNA-377 promoted tube formation in human umbilical vein endothelial cells. We then engineered rat MSCs with lentiviral vectors to either overexpress microRNA-377 (MSC miR-377) or knockdown microRNA-377 (MSC Anti-377) to investigate whether microRNA-377 regulated MSC-induced myocardial angiogenesis, using MSCs infected with lentiviral empty vector to serve as controls (MSC Null). Four weeks after implantation of the microRNA-engineered MSCs into the infarcted rat hearts, the vessel density was significantly increased in MSC Anti-377-hearts, and this was accompanied by reduced fibrosis and improved myocardial function as compared to controls. Adverse effects were observed in MSC miR-377-treated hearts, including reduced vessel density, impaired myocardial function, and increased fibrosis in comparison with MSC Null-group. These findings indicate that hypoxia-responsive microRNA-377 directly targets VEGF in MSCs, and knockdown of endogenous microRNA-377 promotes MSC-induced angiogenesis in the infarcted myocardium. Thus, microRNA-377 may serve as a novel therapeutic target for stem cell-based treatment of ischemic heart disease.
authors.affiliation: Department of Infectious Disease, Nanchang University Medical School, Nanchang, Jiangxi, China; Department of Pathology and Lab Medicine, University of Cincinnati Medical Center, Cincinnati, Ohio, United States of America.
authors.forename: Zhili
authors.initials: Z
authors.lastname: Wen
authors.affiliation: Department of Pathology and Lab Medicine, University of Cincinnati Medical Center, Cincinnati, Ohio, United States of America.
authors.forename: Wei
authors.initials: W
authors.lastname: Huang
authors.affiliation: Department of Pathology and Lab Medicine, University of Cincinnati Medical Center, Cincinnati, Ohio, United States of America.
authors.forename: Yuliang
authors.initials: Y
authors.lastname: Feng
authors.affiliation: Department of Pathology and Lab Medicine, University of Cincinnati Medical Center, Cincinnati, Ohio, United States of America.
authors.forename: Wenfeng
authors.initials: W
authors.lastname: Cai
authors.affiliation: Department of Pathology and Lab Medicine, University of Cincinnati Medical Center, Cincinnati, Ohio, United States of America.
authors.forename: Yuhua
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Pharmacology and Cell Biophysics, University of Cincinnati Medical Center, Cincinnati, Ohio, United States of America.
authors.forename: Xiaohong
authors.initials: X
authors.lastname: Wang
authors.affiliation: Department of Pathology and Lab Medicine, University of Cincinnati Medical Center, Cincinnati, Ohio, United States of America.
authors.forename: Jialiang
authors.initials: J
authors.lastname: Liang
authors.affiliation: Department of Pathology and Lab Medicine, University of Cincinnati Medical Center, Cincinnati, Ohio, United States of America.
authors.forename: Mashhood
authors.initials: M
authors.lastname: Wani
authors.affiliation: Department of Environmental Health, University of Cincinnati Medical Center, Cincinnati, Ohio, United States of America.
authors.forename: Jing
authors.initials: J
authors.lastname: Chen
authors.affiliation: Guangdong Cardiovascular Institute, Guangdong Academy of Medical Sciences, Guangzhou, Guandong, People's Republic of China.
authors.forename: Pin
authors.initials: P
authors.lastname: Zhu
authors.affiliation: Guangdong Cardiovascular Institute, Guangdong Academy of Medical Sciences, Guangzhou, Guandong, People's Republic of China.
authors.forename: Ji-Mei
authors.initials: JM
authors.lastname: Chen
authors.affiliation: Department of Pharmacology and Cell Biophysics, University of Cincinnati Medical Center, Cincinnati, Ohio, United States of America.
authors.forename: Ronald W
authors.initials: RW
authors.lastname: Millard
authors.affiliation: Department of Pharmacology and Cell Biophysics, University of Cincinnati Medical Center, Cincinnati, Ohio, United States of America.
authors.forename: Guo-Chang
authors.initials: GC
authors.lastname: Fan
authors.affiliation: Department of Pathology and Lab Medicine, University of Cincinnati Medical Center, Cincinnati, Ohio, United States of America.
authors.forename: Yigang
authors.initials: Y
authors.lastname: Wang
mesh_terms: D020413:3' Untranslated Regions / Q000235:genetics; D000818:Animals; D015153:Blotting, Western; D015687:Cell Hypoxia; D002478:Cells, Cultured; D004452:Echocardiography; D020869:Gene Expression Profiling; D055785:Gene Knockdown Techniques; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism; D006801:Humans; D008297:Male; D045164:Mesenchymal Stem Cell Transplantation / Q000379:methods; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D035683:MicroRNAs / Q000235:genetics*; D009203:Myocardial Infarction / Q000235:genetics* / Q000503:physiopathology / Q000628:therapy; D009389:Neovascularization, Pathologic / Q000235:genetics*; D018919:Neovascularization, Physiologic / Q000235:genetics; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics* / Q000378:metabolism
chemical_list: D020413:3' Untranslated Regions; C534063:MIRN377 microRNA, human; C000598764:MIRN377 microRNA, rat; D035683:MicroRNAs; D042461:Vascular Endothelial Growth Factor A