Translational Control of Cardiac Excitability

心脏兴奋性的转化控制

• 批准号: 10367512
• 负责人: Gail A Robertson
• 金额: $ 70.4万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-15 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10367512
• 关键词: Action Potentials; Address; Adult; Affinity; Anabolism; Binding Proteins; Biogenesis; Biological Assay; Calcium; Cardiac; Cardiac Myocytes; Cells; Collection; Complex; Coupled; Data; Disease; Electrophysiology (science); Embryo; Equilibrium; Exhibits; Fluorescent in Situ Hybridization; Funding; Heart; Heart failure; Homeostasis; Human; Immunofluorescence Immunologic; Ion Channel; Knowledge; Lead; Macromolecular Complexes; Mass Spectrum Analysis; Mediating; Membrane Potentials; Messenger RNA; Modeling; Multiprotein Complexes; Muscle Cells; Myocardium; Nature; Outcome; Pacemakers; Pathway interactions; Physiological; Proteins; RNA; RNA Interference; RNA-Binding Proteins; Regulation; Reverse Transcriptase Polymerase Chain Reaction; Role; Sarcolemma; Science; Shapes; Sinoatrial Node; Sodium Channel; Specificity; System; Testing; Therapeutic; Tissues; Transcript; Translations; Ventricular; base; heart function; heart rhythm; induced pluripotent stem cell; induced pluripotent stem cell derived cardiomyocytes; innovation; insight; knock-down; macromolecular assembly; novel; protein expression; single molecule; stoichiometry; therapeutic development; translatome

PROJECT SUMMARY/ABSTRACT The regular beating of the heart requires the orchestrated expression of ion channel ensembles specific for ventricular repolarization, pacemaking, conduction and other functions. Recent progress establishes a new way in which channel expression is coordinated. The central hypothesis of this proposal is that a “microtranslatome” of interacting mRNA species encodes functionally related proteins, such as those producing the ventricular action potential. These ion channels assemble cotranslationally into macromolecular complexes that govern higher-order cardiac excitability. The aims of the proposal are to resolve components of cotranslational complexes mediating cardiac repolarization using single-molecule fluorescence in situ hybridization combined with protein immunofluorescence (smFISH/IF) in both embryonic (iPSC) and adult cardiomyocytes. Hypotheses regarding the composition, stoichiometry and cellular localization of these complexes will be tested. Whether such complexes regulate other cardiac functions, such as pacemaking, will be determined. Mechanisms mediating mRNA association, such as candidate RNA binding proteins (RBPs) identified with affinity-capture mass spectrometry, and direct mRNA interactions, will be resolved. These studies are expected to illuminate new mechanisms by which cardiac excitability is controlled and identify novel targets for disease and therapeutic development.

项目概要/摘要 心脏的规律跳动需要特定离子通道群的精心策划的表达 心室复极、起搏、传导等功能的最新进展确立了新的功能。 该提案的中心假设是协调渠道表达的方式。 相互作用的 mRNA 物种的“微翻译组”编码功能相关的蛋白质，例如 产生心室动作电位。 该提案的目的是解决控制高阶心脏兴奋性的复合物。 使用单分子荧光原位介导心脏复极的共翻译复合物 在胚胎 (iPSC) 和成人中进行杂交结合蛋白质免疫荧光 (smFISH/IF) 关于心肌细胞的组成、化学计量和细胞定位的假设。 将测试这些复合物是否调节其他心脏功能，例如起搏功能。 介导 mRNA 关联的机制，例如候选 RNA 结合蛋白 (RBP) 通过亲和捕获质谱法鉴定以及直接的 mRNA 相互作用将得到解决。 预计研究将阐明控制心脏兴奋性的新机制并确定新的机制 疾病和治疗开发的目标。