Mesenchymal stem cells modified with miR-126 release angiogenic factors and activate Notch ligand Delta-like-4, enhancing ischemic angiogenesis and cell survival

Huang,Feng; Zhu,Xiao; Hu,Xin-Qun; Fang,Zhen-Fei; Tang,Liang; Lu,Xiao-Ling; Zhou,Sheng-Hua

doi:10.3892/ijmm.2012.1200

Mesenchymal stem cells modified with miR-126 release angiogenic factors and activate Notch ligand Delta-like-4, enhancing ischemic angiogenesis and cell survival

Authors:
- Feng Huang
- Xiao Zhu
- Xin-Qun Hu
- Zhen-Fei Fang
- Liang Tang
- Xiao-Ling Lu
- Sheng-Hua Zhou
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Affiliations: Department of Cardiology, The Second Xiangya Hospital of Central South University, Changsha, Hunan 410011, P.R. China, Institute of Metabolism and Endocrinology, The Second Xiangya Hospital of Central South University, Changsha, Hunan 410011, P.R. China
Published online on: December 3, 2012 https://doi.org/10.3892/ijmm.2012.1200
Pages: 484-492

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Abstract

The endothelial cell-specific microRNA (miRNA), miR-126, is considered a master regulator of physiological angiogenesis. Transplanted mesenchymal stem cells (MSCs) release soluble factors contributing to neoangiogenesis and cardiac repair. Therefore, we hypothesized that the overexpression of miR-126 may prolong MSC survival and enhance the cell secretome, thereby improving post-infarction angiogenesis and cardiac function. In this study, MSCs harvested from male C57BL/6 mouse bone marrow were infected in vitro with miR-126 (MSCmiR-126) by using recombinant lentiviral vectors; the control cells were either non-transfected or transduced with mock vectors (MSCnull). The results showed an increased secretion of angiogenic factors and a higher resistance against hypoxia in MSCmiR-126 compared with the control cells. The expression of the Notch ligand Delta-like (Dll)-4 in the MSCmiR-126 group was also increased. For in vivo experiments, MSCmiR-126 cultures were intramyocardially injected into the infarct region of the hearts of female C57BL/6 mice (an acute myocardial infarction model) who had undergone ligation of the left anterior descending coronary artery. The survival of MSCmiR-126 cultures, determined by Sry expression, was increased at 7 days after transplantation. MSCmiR-126-treated animals showed significantly improved cardiac function as assessed by echocardiography 2 weeks later. The expression levels of angiogenic factors and Dll-4 in the infarcted myocardium were further increased by MSCmiR-126 compared with MSCs or MSCnull cultures. Furthermore, fluorescent microsphere and histological studies revealed that myocardial blood flow and microvessel density were significantly increased in the MSCmiR-126-transplanted animals. In addition, we found increased immature vessel proliferation following the transplantation of MSCmiR-126 cultures in which the expression of Dll-4 had been knocked down. However, blood vessels with lumen were barely detected, which indicated that Dll-4 plays a key role in tubulogenesis. We conclude that the transplantation of MSCs overexpressing miR-126 can further enhance functional angiogenesis in the ischemic myocardium possibly by the secretion of angiogenic factors and the activation of Dll-4, thus increasing MSC survival. Therefore, MSCs modified with miR-126 may represent a novel and efficient therapeutic approach for ischemic angiogenesis and the improvement of cardiac function.

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