Mitochondrial Calmodulin Kinase II in Physiology and Disease

线粒体钙调蛋白激酶 II 在生理学和疾病中的作用

• 批准号: 8909894
• 负责人: MARK E ANDERSON
• 金额: $ 39.69万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-09 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8909894
• 关键词: ATP Synthesis Pathway; Abbreviations; Agonist; Automobile Driving; Binding; Ca(2+)-Calmodulin Dependent Protein Kinase; Calcium; Calcium/calmodulin-dependent protein kinase; Cardiac Myocytes; Cardiomyopathies; Cell Death; Cells; Cessation of life; Charge; Data; Disease; Dominant-Negative Mutation; Enzymes; Event; Figs - dietary; Genes; Goals; Heart failure; Homeostasis; Infusion procedures; Injury; Inner mitochondrial membrane; Ion Channel; Isoproterenol; Knockout Mice; Mediating; Membrane; Membrane Potentials; Metabolism; Mitochondria; Modeling; Molecular; Molecular Target; Mus; Myocardial; Myocardial dysfunction; Myocardium; NADH; Nature; Neonatal; Oxidative Phosphorylation; Oxidoreductase; Pathway interactions; Permeability; Pharmaceutical Preparations; Phosphorylation; Physiological; Physiology; Production; Proteins; Public Health; Reactive Oxygen Species; Resistance; Role; Second Messenger Systems; Serine; Site; Source; Stimulus; Stress; Testing; Time; Transgenic Organisms; Work; base; biophysical properties; calmodulin-dependent protein kinase II; clinically relevant; effective therapy; in vivo; innovation; knock-down; mouse model; prevent; public health relevance; response; second messenger; therapeutic target

DESCRIPTION (provided by applicant): Heart failure is a major public health problem without adequate therapies. Loss of myocardial Ca2+ homeostasis and mitochondrial Ca2+ overload are fundamental events driving heart failure progression, but no currently available therapies prevent excessive mitochondrial Ca2+ entry. In 2011, after a 50 year search, two groups independently identified the molecular basis for the mitochondrial Ca2+ uniporter (MCU), the main pathway for Ca2+ entry into mitochondria. We developed new, myocardial-selective transgenic and inducible knock out mouse models of MCU inhibition to test this concept in vivo. Our new mice with myocardial MCU inhibition are viable and our preliminary data show they are resistant to myocardial death after isoproterenol infusion. Here we propose to establish how MCU inhibition contributes to myocardial physiology and disease. The multifunctional Ca2+ and calmodulin-dependent protein kinase II (CaMKII) contributes to heart failure by promoting defective intracellular Ca2+ handling, including mitochondrial Ca2+ overload, but the molecular targets for cardiomyopathic actions of CaMKII are uncertain. During the original period of this competing renewal, we found that CaMKII is present in mitochondria, that mitochondrial CaMKII inhibition reduces MCU- mediated mitochondrial Ca2+ entry and protects against mitochondrial Ca2+ overload in clinically-relevant models of heart failure (Joiner Nature 2012). We identified key sites on MCU (serines 57 and 92) that are essential for CaMKII agonist actions. Thus, MCU is the first validated CaMKII target protein in mitochondria. Here we propose to test the role of mitochondrial CaMKII at MCU in myocardial physiology and disease. The overall goal of this competitive renewal application is to determine the importance of MCU and mitochondrial CaMKII-dependent MCU phosphorylation for myocardial metabolism and disease using 3 specific aims. 1. Determine the effect of MCU inhibition on myocardial physiology; 2. Determine the effect of MCU inhibition on myocardial responses to pathological stress; 3. Determine the effects of CaMKII-dependent MCU phosphorylation on metabolism and disease.

描述（由申请人提供）：心力衰竭是一个没有适当治疗的重大公共卫生问题。心肌Ca2+稳态的丧失和线粒体Ca2+超载是驱动心力衰竭进展的基本事件，但目前没有可用的治疗方法防止过量的线粒体Ca2+进入。2011年，经过50年的研究，两个研究小组独立确定了线粒体Ca2+单转运体（MCU）的分子基础，这是Ca2+进入线粒体的主要途径。我们开发了新的心肌选择性转基因和诱导敲除小鼠模型，以在体内验证这一概念。我们的新小鼠心肌MCU抑制是有活力的，我们的初步数据显示异丙肾上腺素输注后它们对心肌死亡具有抵抗性。在这里，我们提出建立MCU抑制如何有助于心肌生理和疾病。多功能Ca2+和钙调素依赖性蛋白激酶II （CaMKII）通过促进细胞内Ca2+处理缺陷，包括线粒体Ca2+超载，有助于心力衰竭，但CaMKII的心肌病作用的分子靶点是不确定的。在这种竞争性更新的初始阶段，我们发现CaMKII存在于线粒体中，线粒体CaMKII抑制减少MCU介导的线粒体Ca2+进入，并在临床相关的心力衰竭模型中保护线粒体Ca2+过载（Joiner Nature 2012）。我们确定了MCU上的关键位点（丝氨酸57和92）是CaMKII激动剂作用所必需的。因此，MCU是线粒体中第一个被证实的CaMKII靶蛋白。在这里，我们提议测试线粒体CaMKII在MCU中在心肌生理和疾病中的作用。这项竞争性更新申请的总体目标是确定MCU和线粒体camkii依赖的MCU磷酸化对心肌代谢和疾病的重要性。1. 确定MCU抑制对心肌生理的影响；2. 测定MCU抑制对心肌病理应激反应的影响；3. 确定camkii依赖性MCU磷酸化对代谢和疾病的影响。