Direct and Indirect Involvement of MicroRNA-499 in Clinical and Experimental Cardiomyopathy

Rationale: MicroRNA-499 and other members of the myomiR family regulate myosin isoforms in pressure-overload hypertrophy. miR-499 expression varies in human disease, but results of mouse cardiac miR-499 overexpression are inconsistent, either protecting against ischemic damage or aggravating cardiomyopathy after pressure overload. Likewise, there is disagreement over direct and indirect cardiac mRNAs targeted in vivo by miR-499. Objective: To define the associations between regulated miR-499 level in clinical and experimental heart disease and modulation of its predicted mRNA targets and to determine the consequences of increased cardiac miR-499 on direct mRNA targeting, indirect mRNA modulation, and on myocardial protein content and posttranslational modification. Methods and Results: miR-499 levels were increased in failing and hypertrophied human hearts and associated with decreased levels of predicted target mRNAs. Likewise, miR-499 is increased in Gq-mediated murine cardiomyopathy. Forced cardiomyocyte expression of miR-499 at levels comparable to human cardiomyopathy induced progressive murine heart failure and exacerbated cardiac remodeling after pressure overloading. Genome-wide RNA-induced silencing complex and RNA sequencing identified 67 direct, and numerous indirect, cardiac mRNA targets, including Akt and MAPKs. Myocardial proteomics identified alterations in protein phosphorylation linked to the miR-499 cardiomyopathy phenotype, including of heat shock protein 90 and protein serine/threonine phosphatase 1-&agr;. Conclusions: miR-499 is increased in human and murine cardiac hypertrophy and cardiomyopathy, is sufficient to cause murine heart failure, and accelerates maladaptation to pressure overloading. The deleterious effects of miR-499 reflect the cumulative consequences of direct and indirect mRNA regulation, modulation of cardiac kinase and phosphatase pathways, and higher-order effects on posttranslational modification of myocardial proteins.

[1]  D. Brautigan,et al.  Protein Ser/ Thr phosphatases – the ugly ducklings of cell signalling , 2013, The FEBS journal.

[2]  R. Danziger,et al.  Non-histone lysine acetylated proteins in heart failure. , 2012, Biochimica et biophysica acta.

[3]  G. Dorn,et al.  A Human 3′ miR-499 Mutation Alters Cardiac mRNA Targeting and Function , 2012, Circulation research.

[4]  L. Neckers,et al.  Post-translational modifications of Hsp90 and their contributions to chaperone regulation. , 2012, Biochimica et biophysica acta.

[5]  S. Fichtlscherer,et al.  Circulating microRNAs: biomarkers or mediators of cardiovascular diseases? , 2011, Arteriosclerosis, thrombosis, and vascular biology.

[6]  E. Olson,et al.  Therapeutic Inhibition of miR-208a Improves Cardiac Function and Survival During Heart Failure , 2011, Circulation.

[7]  G. Dorn,et al.  miR-15 Family Regulates Postnatal Mitotic Arrest of Cardiomyocytes , 2011, Circulation research.

[8]  D. Bartel,et al.  Weak Seed-Pairing Stability and High Target-Site Abundance Decrease the Proficiency of lsy-6 and Other miRNAs , 2011, Nature Structural &Molecular Biology.

[9]  D. Srivastava,et al.  Elevated miR-499 Levels Blunt the Cardiac Stress Response , 2011, PloS one.

[10]  E. Saridakis,et al.  Oxidation State-dependent Protein-Protein Interactions in Disulfide Cascades , 2011, The Journal of Biological Chemistry.

[11]  Derek J Van Booven,et al.  RISC RNA Sequencing for Context-Specific Identification of In Vivo MicroRNA Targets , 2011, Circulation research.

[12]  S. Nef,et al.  The Molecular Chaperone Hsp90α Is Required for Meiotic Progression of Spermatocytes beyond Pachytene in the Mouse , 2010, PloS one.

[13]  G. Dorn,et al.  Receptor-Independent Cardiac Protein Kinase C&agr; Activation by Calpain-Mediated Truncation of Regulatory Domains , 2010, Circulation research.

[14]  A. Newton,et al.  PHLPP-1 Negatively Regulates Akt Activity and Survival in the Heart , 2010, Circulation research.

[15]  Qingbo Xu,et al.  Short Communication: Asymmetric Dimethylarginine Impairs Angiogenic Progenitor Cell Function in Patients With Coronary Artery Disease Through a MicroRNA-21–Dependent Mechanism , 2010, Circulation research.

[16]  Nicholas T. Ingolia,et al.  Mammalian microRNAs predominantly act to decrease target mRNA levels , 2010, Nature.

[17]  Derek J Van Booven,et al.  Deep mRNA Sequencing for In Vivo Functional Analysis of Cardiac Transcriptional Regulators: Application to G&agr;q , 2010, Circulation research.

[18]  Jiahuai Han,et al.  Specific Regulation of Noncanonical p38&agr; Activation by Hsp90-Cdc37 Chaperone Complex in Cardiomyocyte , 2010, Circulation research.

[19]  E. Olson,et al.  MicroRNA regulatory networks in cardiovascular development. , 2010, Developmental cell.

[20]  Yixue Li,et al.  Regulation of Cellular Metabolism by Protein Lysine Acetylation , 2010, Science.

[21]  L. Leinwand,et al.  Uncoupling of Expression of an Intronic MicroRNA and Its Myosin Host Gene by Exon Skipping , 2010, Molecular and Cellular Biology.

[22]  J. Nerbonne,et al.  MicroRNA-133a Protects Against Myocardial Fibrosis and Modulates Electrical Repolarization Without Affecting Hypertrophy in Pressure-Overloaded Adult Hearts , 2010, Circulation research.

[23]  E. Olson,et al.  A family of microRNAs encoded by myosin genes governs myosin expression and muscle performance. , 2009, Developmental cell.

[24]  P. Lasko,et al.  Hsp90 regulates the function of argonaute 2 and its recruitment to stress granules and P-bodies. , 2009, Molecular biology of the cell.

[25]  L. Pachter,et al.  TopHat: discovering splice junctions with RNA-Seq , 2009, Bioinform..

[26]  Derek J Van Booven,et al.  Reciprocal Regulation of Myocardial microRNAs and Messenger RNA in Human Cardiomyopathy and Reversal of the microRNA Signature by Biomechanical Support , 2009, Circulation.

[27]  S. Soddu,et al.  HIPKs: Jack of all trades in basic nuclear activities. , 2008, Biochimica et biophysica acta.

[28]  D. Bartel,et al.  The impact of microRNAs on protein output , 2008, Nature.

[29]  M. Stephens,et al.  RNA-seq: an assessment of technical reproducibility and comparison with gene expression arrays. , 2008, Genome research.

[30]  B. Williams,et al.  Mapping and quantifying mammalian transcriptomes by RNA-Seq , 2008, Nature Methods.

[31]  M. Hecker,et al.  A proteome map of murine heart and skeletal muscle , 2008, Proteomics.

[32]  F. Veglia,et al.  Oxidized proteins in plasma of patients with heart failure: Role in endothelial damage , 2008, European journal of heart failure.

[33]  G. Dorn,et al.  Nix-Mediated Apoptosis Links Myocardial Fibrosis, Cardiac Remodeling, and Hypertrophy Decompensation , 2008, Circulation.

[34]  Roy Parker,et al.  The Two Faces of miRNA , 2007, Science.

[35]  Zhenyu Xuan,et al.  A biochemical approach to identifying microRNA targets , 2007, Proceedings of the National Academy of Sciences.

[36]  Richard J Jackson,et al.  MicroRNAs repress translation of m7Gppp-capped target mRNAs in vitro by inhibiting initiation and promoting deadenylation. , 2007, Genes & development.

[37]  Xiaoxia Qi,et al.  Control of Stress-Dependent Cardiac Growth and Gene Expression by a MicroRNA , 2007, Science.

[38]  Thomas Thum,et al.  MicroRNAs in the Human Heart: A Clue to Fetal Gene Reprogramming in Heart Failure , 2007, Circulation.

[39]  J. Buchner,et al.  The phosphatase Ppt1 is a dedicated regulator of the molecular chaperone Hsp90 , 2006, The EMBO journal.

[40]  Martin Kuiper,et al.  BiNGO: a Cytoscape plugin to assess overrepresentation of Gene Ontology categories in Biological Networks , 2005, Bioinform..

[41]  D. Chuang,et al.  Crystal structure of pyruvate dehydrogenase kinase 3 bound to lipoyl domain 2 of human pyruvate dehydrogenase complex , 2005, The EMBO journal.

[42]  Andrew N. Carr,et al.  Enhancement of Cardiac Function and Suppression of Heart Failure Progression By Inhibition of Protein Phosphatase 1 , 2005, Circulation research.

[43]  G. Dorn,et al.  Physiological Growth Synergizes With Pathological Genes in Experimental Cardiomyopathy , 2004, Circulation research.

[44]  P. Csermely,et al.  Hsp90 isoforms: functions, expression and clinical importance , 2004, FEBS letters.

[45]  D. Schulze,et al.  Sodium/Calcium Exchanger (NCX1) Macromolecular Complex* , 2003, Journal of Biological Chemistry.

[46]  A. Brown,et al.  Role of the Cytosolic Chaperones Hsp70 and Hsp90 in Maturation of the Cardiac Potassium Channel hERG , 2003, Circulation research.

[47]  G. Dorn,et al.  Phenotyping hypertrophy: eschew obfuscation. , 2003, Circulation research.

[48]  N. Rosen,et al.  Akt Forms an Intracellular Complex with Heat Shock Protein 90 (Hsp90) and Cdc37 and Is Destabilized by Inhibitors of Hsp90 Function* , 2002, The Journal of Biological Chemistry.

[49]  S. Reiken,et al.  Protein Kinase A and Two Phosphatases Are Components of the Inositol 1,4,5-Trisphosphate Receptor Macromolecular Signaling Complex* , 2002, The Journal of Biological Chemistry.

[50]  F. Speleman,et al.  Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes , 2002, Genome Biology.

[51]  A. Ishida,et al.  Involvement of Hsp90 in Signaling and Stability of 3-Phosphoinositide-dependent Kinase-1* , 2002, The Journal of Biological Chemistry.

[52]  S. Marx,et al.  Phosphorylation-Dependent Regulation of Ryanodine Receptors , 2001, The Journal of cell biology.

[53]  G. Varani,et al.  The G x U wobble base pair. A fundamental building block of RNA structure crucial to RNA function in diverse biological systems. , 2000, EMBO reports.

[54]  John W. Adams,et al.  Enhanced Galphaq signaling: a common pathway mediates cardiac hypertrophy and apoptotic heart failure. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[55]  G. Dorn,et al.  Transgenic Gαq overexpression induces cardiac contractile failure in mice , 1997 .

[56]  P. Greengard,et al.  Phosphorylation and inactivation of protein phosphatase 1 by cyclin-dependent kinases. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[57]  Yanrui Li,et al.  miR-499 regulates mitochondrial dynamics by targeting calcineurin and dynamin-related protein-1 , 2011, Nature Medicine.

[58]  Roy Parker,et al.  Molecular biology. The two faces of miRNA. , 2007, Science.

[59]  E. Olson,et al.  Balancing contractility and energy production: the role of myocyte enhancer factor 2 (MEF2) in cardiac hypertrophy. , 2004, Recent progress in hormone research.

[60]  S. Miyamoto,et al.  Cardiomyocyte calcium and calcium/calmodulin-dependent protein kinase II: friends or foes? , 2004, Recent progress in hormone research.