The specific roles of miRNAs in vascular function are just beginning to be explored. Our previous work has shown that reduction of overall miRNA levels via Dicer silencing in human endothelial cells (EC) strongly regulates angiogenic gene expression and impairs aspects of in vitro angiogenesis including EC growth and morphogenesis. These in vitro experiments are supported by new exciting preliminary data in vivo data using mice conditionally lacking the rate limiting enzyme (Dicer) in miRNA synthesis in EC. Mice lacking Dicer in EC (EC specific Dicer KO) are viable but exhibit impaired post-natal angiogenic responses. In order to dissect the relationships between signal transduction, gene expression and miRNAs in EC, we show that treatment of EC with VEGF stimulates time-dependent changes in global miRNA profiles and show that components of the miRNA cluster, miR 17-92 can regulate aspects of VEGF induced cell growth and morphogenesis. In addition, we have identified a specific miRNA (miR-155) that regulates the levels of endothelial nitric oxide synthase (eNOS) in cultured EC. Thus, we hypothesize that Dicer generated endothelial miRNAs regulate the extent of angiogenesis and blood flow control in vivo by regulating mRNA levels and/or the translational stability of important proteins. We will: 1. Identify and characterize angiogenic growth factor regulated miRNAs in EC;2. Define the roles of miR 17-92 in models of angiogenesis and 3. Define the roles of miR-155 in regulating eNOS function in vitro and in vivo. Collectively, this work will facilitate the understanding of the importance of miRNAs in EC biology and shed insights into their role as potential therapeutics targets.

Public Health Relevance

This research is relevant to public health since endothelial dysfunction is a common manifestation of most cardiovascular diseases. Our research has discovered the major mechanisms of how the endothelium control blood flow and atherogenesis. Research supported by this grant may help identify new drugs that reduce heart disease and improve the quality of life of people suffering with cardiovascular disease.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
5R01HL096670-03
Application #
8245750
Study Section
Vascular Cell and Molecular Biology Study Section (VCMB)
Program Officer
Gao, Yunling
Project Start
2010-04-06
Project End
2014-03-31
Budget Start
2012-04-01
Budget End
2013-03-31
Support Year
3
Fiscal Year
2012
Total Cost
$470,224
Indirect Cost
$182,052
Name
Yale University
Department
Pharmacology
Type
Schools of Medicine
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520
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