The non-hypertrophic role of calcineurin in regulating cardiac structure-function

钙调神经磷酸酶在调节心脏结构功能中的非肥厚作用

基本信息

批准号：
8012835
负责人：
Jennifer Michelle Davis
金额：
$ 5.3万
依托单位：
CINCINNATI CHILDRENS HOSP MED CTR
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-12-15 至 2011-12-14
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8012835
关键词：
Acute Adrenergic Agents Adult American Biology Calcineurin Calcineurin inhibitor Cardiac Cardiac Myocytes Cardiovascular Diseases Cell physiology Cells Cessation of life Coupling Critical Pathways Development Diagnosis Gene Expression Gene Transfer Genetic Genetic Models Genetic Transcription Grant Growth Heart Heart Diseases Heart failure Hypertrophy In Vitro Investigation Language Left Ventricular Function Measurement Mediating Molecular Morphology Mus Muscle Cells Muscle function Myocardium Outcome Pathologic Pathway interactions Performance Phosphoric Monoester Hydrolases Phosphorylation Physiological Play Process Protein phosphatase Proteins Proteomics Rattus Regulation Relaxation Reporting Role Signal Transduction Small Interfering RNA Staging Stress Structure Testing Tissues Transgenes Ventricular Ventricular Function Ventricular Remodeling Work adrenergic comparative design functional outcomes in vivo inhibitor/antagonist innovation insight mouse model new growth novel therapeutics premature response ventricular hypertrophy

项目摘要

DESCRIPTION (provided by applicant): Over 5 million Americans are diagnosed with heart failure yearly. Heart failure is generally accompanied by pathologic growth of the heart. Calcineurin signaling is an essential regulator of the hypertrophic response. A new non-hypertrophic role for calcineurin is emerging, in which calcineurin might directly maintain viability and cardiac performance, as mice with cardiac-specific calcineurin deletion have normal cardiac morphology but poor left ventricular function and premature death. To date the direct versus compensatory effects of calcineurin deletion on cardiac function remain unresolved, and reports show both positive and negative functional outcomes in calcineurin-deleted cardiac muscle. Substantial alterations in Ca2+ handling gene expression were identified in a screen of calcineurin deficient hearts. This suggests that calcineurin may regulate Ca2+ cycling proteins, which are critical functional components, as fluctuations in cytosolic Ca2+ initiate contraction and relaxation. Thus, this proposal seeks to test the hypothesis that calcineurin is directly regulating hypertrophy-independent cardiac viability and ventricular performance through post-translational and NFAT-dependent regulation of Ca2+ handling. This grant features a comparative analysis of acute gene transfer of calcineurin-targeted siRNA to cardiac myocytes and a conditional Lox-P targeted calcineurin mouse model to elucidate the direct versus indirect effects of calcineurin loss on cardiac myocyte function in vitro (aiml), and to determine the phosphorylation status of Ca2+ handling proteins (aim 2). Myocyte contractility will be examined in isolated cardiac myocytes from both genetic models using high fidelity cell shortening and Ca2+ measurements, while candidate protein and proteomic analysis will determine the phosphorylation status of Ca2+ handling proteins. A genetic rescue strategy will also be used to investigate if NFAT-dependent transcription underlies the poor survivability in calcineurin deficient mice (aim 3). This proposal should make substantial contributions to elucidating calcineurin's additional regulatory roles in the heart thereby laying the ground work for developing new therapeutic strategies for heart failure. Lay Language: Calcineurin-dependent pathologic growth of the heart is associated with end-stage heart failure. A new growth-independent regulatory role for calcineurin in cardiac function is now emerging that suggests calcineurin deletion can directly alter cardiac muscle function. This proposal is designed to elucidate the direct versus indirect effects of calcineurin on cardiac function independent of cardiac growth at the cellular and organismal level. The outcome of this work should make significant inroads into designing molecular heart failure therapies with translational potential.

描述（由申请人提供）：每年被诊断出超过500万美国人的心力衰竭。心力衰竭通常伴随着心脏的病理生长。钙调蛋白信号传导是肥厚反应的重要调节剂。钙调蛋白酶的新型非嗜性作用正在出现，其中钙调神经蛋白可能直接保持生存能力和心脏表现，因为具有心脏特异性钙调神经磷酸缺失的小鼠具有正常的心脏形态，但左心室功能和过早死亡较差。迄今为止，钙调蛋白缺失对心脏功能的直接与补偿性影响尚未解决，报告显示钙调蛋白缺失的心脏肌肉的阳性和负功能结果。 Ca2+处理基因表达的实质变化在钙调神经蛋白缺乏心脏的筛选中鉴定出来。这表明钙调蛋白可以调节Ca2+循环蛋白，这些蛋白是关键功能成分，因为胞质Ca2+启动收缩和放松的波动。因此，该提案旨在检验钙调神经蛋白直接通过翻译后和NFAT依赖性CA2+处理的调节来调节骨蛋白直接调节肥大依赖肥大的心脏活力和心室性能的假设。该赠款具有比较分析钙调神经素靶向心肌细胞的急性基因转移以及有条件的LOX-P靶向钙调蛋白小鼠模型的比较分析，以阐明钙调神经蛋白损失对心脏肌细胞功能对体外功能的直接和间接影响（AIML），并确定CA2+ protsing protee的磷酸化状态（AIML）（AIML）。使用高富达细胞缩短和Ca2+测量值，将在遗传模型的孤立心肌细胞中检查肌细胞收缩力，而候选蛋白和蛋白质组学分析将确定Ca2+处理蛋白的磷酸化状态。遗传救援策略还将用于研究NFAT依赖性转录是否是钙调神经磷酸酶缺乏小鼠的生存能力较差的基础（AIM 3）。该提案应为阐明钙调神经素在心脏中的其他调节作用做出重大贡献，从而为开发新的心力衰竭治疗策略奠定了基础工作。外行语言：心脏依赖钙调神经素的病理生长与终末期心力衰竭有关。现在正在出现钙调神经蛋白酶在心脏功能中的一种新的非生长调节作用，这表明钙调神经磷酸酶缺失可以直接改变心脏肌肉功能。该建议旨在阐明钙调神经蛋白对心脏功能的直接与间接作用，而与细胞和有机水平的心脏生长无关。这项工作的结果应大大涉及设计具有转化潜力的分子心力衰竭疗法。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Heart-specific deletion of CnB1 reveals multiple mechanisms whereby calcineurin regulates cardiac growth and function.

DOI：
10.1074/jbc.m109.056143
发表时间：
2010-02-26
期刊：
The Journal of biological chemistry
影响因子：
0
作者：
Maillet M;Davis J;Auger-Messier M;York A;Osinska H;Piquereau J;Lorenz JN;Robbins J;Ventura-Clapier R;Molkentin JD
通讯作者：
Molkentin JD