microRNAs (miRs) are small non-coding RNAs that regulate protein expression by destabilization and/or translational inhibition of target messenger RNAs (mRNAs). Like mRNAs, expression of miRs is regulated in cardiac hypertrophy and heart failure, fine-tuning cardiomyocyte function in response to immediate physiological demands. Single miRs tend to regulate numerous effectors within the same functional pathway, producing a coherent physiological response via multiple parallel perturbations, and are therefore attractive therapeutic targets. However, targeting of dozens or hundreds of different mRNAs by one miR, together with regulated expression of miRs and mRNAs in cardiac disease, complicates delineation of specific miR functions relevant to heart disease. We have developed new techniques combining RNA sequencing on massively parallel next generation platforms with in vivo miR programming of cardiac RNA-induced signaling complexes to overcome this problem. Additionally, we have identified common sequence variants and rare mutations within the mature miR sequence of 30 human miRs and demonstrated that both seed sequence and non-seed sequence miR variants can alter mRNA targeting. Accordingly, we hypothesize that miR effects on the normal and diseased heart are determined by the levels of expressed miRs, the levels of expressed mRNAs, and the efficiency of miR-mRNA pairing as determined by sequence complementarity, and that miR sequence variations further alter miR effects, independent of miR and mRNA expression levels. To examine these hypotheses we propose the first studies to systematically and comprehensively define important miR-mRNA pairing events and determine their functional consequences in genetically programmed mouse hearts under different pathophysiological conditions. We will follow with studies to determining the consequences of human miR sequence variants and mutations on cardiac miR-mRNA targeting, target protein expression, and cardiac structure and function. We are uniquely positioned to achieve these goals by combining our expertise in human genomics, RNA sequencing, and generation and analysis of murine genetic models. It is our long-term goal to integrate studies of human gene variation and cardiac disease with in vitro cell-based and in vivo murine experimental systems to fully understand the impact of miRs on the heart.

Public Health Relevance

microRNAs (miRs) represent an entirely new and unexpected level of biological regulation, targeting mRNA transcripts for degradation or suppression. miRs are dynamically regulated in human cardiac disease. We have found that miR sequence variations, which will affect their binding to and targeting of mRNAs, are surprisingly common in human subjects. Here, we propose studies to define cardiac miR-mRNA interactions in order to better understand miR biology in normal and diseased hearts. These studies will employ novel but fully validated next generation sequencing technologies in combination with state-of-the art in vitro and in vivo functional modeling to translate the clinical observations into mechanistic insights.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Project (R01)
Project #
1R01HL108943-01A1
Application #
8238967
Study Section
Cardiac Contractility, Hypertrophy, and Failure Study Section (CCHF)
Program Officer
Adhikari, Bishow B
Project Start
2011-12-15
Project End
2015-11-30
Budget Start
2011-12-15
Budget End
2012-11-30
Support Year
1
Fiscal Year
2012
Total Cost
$380,000
Indirect Cost
$130,000
Name
Washington University
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
068552207
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
Dorn 2nd, Gerald W (2016) Central Parkin: The evolving role of Parkin in the heart. Biochim Biophys Acta 1857:1307-12
Song, Moshi; Matkovich, Scot J; Zhang, Yan et al. (2015) Combined cardiomyocyte PKCδ and PKCε gene deletion uncovers their central role in restraining developmental and reactive heart growth. Sci Signal 8:ra39
Matkovich, Scot J; Dorn 2nd, Gerald W; Grossenheider, Tiffani C et al. (2015) Cardiac Disease Status Dictates Functional mRNA Targeting Profiles of Individual MicroRNAs. Circ Cardiovasc Genet 8:774-84
Dorn 2nd, Gerald W (2015) Cardiac regeneration - Alchemy, science, and a wee bit of magic? J Mol Cell Cardiol 81:10-1
Matkovich, Scot J; Dorn 2nd, Gerald W (2015) Deep sequencing of cardiac microRNA-mRNA interactomes in clinical and experimental cardiomyopathy. Methods Mol Biol 1299:27-49
Gong, Guohua; Song, Moshi; Csordas, Gyorgy et al. (2015) Parkin-mediated mitophagy directs perinatal cardiac metabolic maturation in mice. Science 350:aad2459
Devaux, Yvan; Zangrando, Jennifer; Schroen, Blanche et al. (2015) Long noncoding RNAs in cardiac development and ageing. Nat Rev Cardiol 12:415-25
Dorn 2nd, Gerald W; Matkovich, Scot J (2015) Epitranscriptional regulation of cardiovascular development and disease. J Physiol 593:1799-808
Dorn 2nd, Gerald W (2014) LIPCAR: a mitochondrial lnc in the noncoding RNA chain? Circ Res 114:1548-50
Matkovich, Scot J; Edwards, John R; Grossenheider, Tiffani C et al. (2014) Epigenetic coordination of embryonic heart transcription by dynamically regulated long noncoding RNAs. Proc Natl Acad Sci U S A 111:12264-9

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