title: MicroRNA-1 and -133 increase arrhythmogenesis in heart failure by dissociating phosphatase activity from RyR2 complex.
abstract: In heart failure (HF), arrhythmogenic spontaneous sarcoplasmic reticulum (SR) Ca(2+) release and afterdepolarizations in cardiac myocytes have been linked to abnormally high activity of ryanodine receptors (RyR2s) associated with enhanced phosphorylation of the channel. However, the specific molecular mechanisms underlying RyR2 hyperphosphorylation in HF remain poorly understood. The objective of the current study was to test the hypothesis that the enhanced expression of muscle-specific microRNAs (miRNAs) underlies the HF-related alterations in RyR2 phosphorylation in ventricular myocytes by targeting phosphatase activity localized to the RyR2. We studied hearts isolated from canines with chronic HF exhibiting increased left ventricular (LV) dimensions and decreased LV contractility. qRT-PCR revealed that the levels of miR-1 and miR-133, the most abundant muscle-specific miRNAs, were significantly increased in HF myocytes compared with controls (2- and 1.6-fold, respectively). Western blot analyses demonstrated that expression levels of the protein phosphatase 2A (PP2A) catalytic and regulatory subunits, which are putative targets of miR-133 and miR-1, were decreased in HF cells. PP2A catalytic subunit mRNAs were validated as targets of miR-133 by using luciferase reporter assays. Pharmacological inhibition of phosphatase activity increased the frequency of diastolic Ca(2+) waves and afterdepolarizations in control myocytes. The decreased PP2A activity observed in HF was accompanied by enhanced Ca(2+)/calmodulin-dependent protein kinase (CaMKII)-mediated phosphorylation of RyR2 at sites Ser-2814 and Ser-2030 and increased frequency of diastolic Ca(2+) waves and afterdepolarizations in HF myocytes compared with controls. In HF myocytes, CaMKII inhibitory peptide normalized the frequency of pro-arrhythmic spontaneous diastolic Ca(2+) waves. These findings suggest that altered levels of major muscle-specific miRNAs contribute to abnormal RyR2 function in HF by depressing phosphatase activity localized to the channel, which in turn, leads to the excessive phosphorylation of RyR2s, abnormal Ca(2+) cycling, and increased propensity to arrhythmogenesis.
authors.affiliation: The Davis Heart and Lung Research Institute, The Ohio State University, Columbus, Ohio, United States of America.
authors.forename: Andriy E
authors.initials: AE
authors.lastname: Belevych
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Sansom
authors.forename: Radmila
authors.initials: R
authors.lastname: Terentyeva
authors.forename: Hsiang-Ting
authors.initials: HT
authors.lastname: Ho
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Nishijima
authors.forename: Mickey M
authors.initials: MM
authors.lastname: Martin
authors.forename: Hitesh K
authors.initials: HK
authors.lastname: Jindal
authors.forename: Jennifer A
authors.initials: JA
authors.lastname: Rochira
authors.forename: Yukiko
authors.initials: Y
authors.lastname: Kunitomo
authors.forename: Maha
authors.initials: M
authors.lastname: Abdellatif
authors.forename: Cynthia A
authors.initials: CA
authors.lastname: Carnes
authors.forename: Terry S
authors.initials: TS
authors.lastname: Elton
authors.forename: Sandor
authors.initials: S
authors.lastname: Györke
authors.forename: Dmitry
authors.initials: D
authors.lastname: Terentyev
mesh_terms: D000818:Animals; D001145:Arrhythmias, Cardiac / Q000235:genetics* / Q000378:metabolism*; D002118:Calcium / Q000378:metabolism; D002384:Catalysis; D020134:Catalytic Domain; D004285:Dogs; D004594:Electrophysiology / Q000379:methods; D017930:Genes, Reporter; D006333:Heart Failure / Q000150:complications / Q000378:metabolism; D006352:Heart Ventricles / Q000473:pathology; D006801:Humans; D007545:Isoproterenol / Q000494:pharmacology; D035683:MicroRNAs / Q000378:metabolism*; D008954:Models, Biological; D032342:Muscle Cells / Q000378:metabolism; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*; D010766:Phosphorylation; D012333:RNA, Messenger / Q000378:metabolism; D011943:Receptors, Adrenergic, beta / Q000378:metabolism; D019837:Ryanodine Receptor Calcium Release Channel / Q000378:metabolism*; D012519:Sarcoplasmic Reticulum / Q000378:metabolism
chemical_list: C544547:MIRN1 microRNA, human; C548234:MIRN133 microRNA, human; D035683:MicroRNAs; D012333:RNA, Messenger; D011943:Receptors, Adrenergic, beta; D019837:Ryanodine Receptor Calcium Release Channel; D010744:Phosphoric Monoester Hydrolases; D007545:Isoproterenol; D002118:Calcium