Elucidation of the Role of a Novel Cardiac Micropeptide in the Control of Heart Function and Dysfunction

阐明新型心脏微肽在控制心脏功能和功能障碍中的作用

基本信息

批准号：
9205181
负责人：
Catherine A Makarewich
金额：
$ 5.71万
依托单位：
UT SOUTHWESTERN MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-02-01 至 2019-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9205181
关键词：
Amino Acids Animals Binding Biochemical Biological Assay Ca(2+)-Transporting ATPase Calcineurin Calcium Cardiac Cardiac Myocytes Cardiovascular Diseases Cause of Death Cells Cessation of life Characteristics Chronic Clinical Confocal Microscopy Coupling DNA Sequence Defect Depressed mood Developed Countries Developing Countries Development Dilated Cardiomyopathy Disease Disease Progression Disease model Disease susceptibility Electron Microscopy Epidemic Event Expenditure Financial compensation Genes Goals Healthcare Heart Heart Abnormalities Heart Diseases Heart failure Homeostasis Human Human Characteristics Impairment In Vitro Integral Membrane Protein Knockout Mice Lead Mass Spectrum Analysis Measurement Mediating Membrane Methodology Molecular Morbidity - disease rate Mus Muscle Muscle Cells Muscle relaxation phase Myocardial dysfunction Names Open Reading Frames PPP3CA gene Pathologic Pathologic Processes Patients Peptides Performance Physiological Physiological Processes Plant Genome Prevalence Property Protein phosphatase Proteins Public Health Pump RNA Regulation Regulatory Pathway Relaxation Role Sarcoplasmic Reticulum Sequence Homology Signal Transduction Skeletal Muscle Structure Systole Systolic heart failure Testing Therapeutic Untranslated RNA Yeasts adeno-associated viral vector base disability gene therapy genetic approach heart function in vivo inhibitor/antagonist insight mortality neglect new therapeutic target novel novel therapeutics overexpression phospholamban pressure public health relevance response sarcolipin yeast two hybrid system

项目摘要

DESCRIPTION (provided by applicant): Cardiovascular disease (CVD) is the leading cause of death and disability in industrialized nations and its prevalence is rising rapidly in developing nations. In particular, heart failure (HF) has been singled out as an epidemic and is a staggering clinical and public health problem associated with significant morbidity, mortality, and healthcare expenditures. Current therapies only delay morbidity and mortality in patients with chronic systolic heart failure, as disease progression typically continues unabated resulting in death. Novel therapies for heart failure are desperately needed, which will require a greater understanding of the underlying molecular mechanisms that underlie this disease. A universal characteristic of the failing heart is impaired Ca2+ cycling through the sarcoplasmic reticulum (SR), the major internal store of Ca2+ in cardiomyocytes. Indeed, cardiomyocytes from failing hearts consistently show defective excitation contraction-coupling characterized by diminished SR Ca2+ sequestration and decreased intracellular Ca2+ transients, events that contribute to impaired contractility and relaxation that culminate in depressed pumping action of the heart. Ca2+ re- sequestration into the SR is mediated by a Ca2+-ATPase (SERCA), whose activity is known to be reversibly regulated by the small integral membrane proteins phospholamban (PLN) and sarcolipin (SLN). Enhancing SERCA activity and function has been suggested as an important clinical approach for treating HF by loading the SR with greater Ca2+ levels to augment Ca2+ release resulting in greater myocyte contractility during systole. Apart from gene therapy to actually replace SERCA protein (which is currently in human clinical trails with an AAV vector), we currently lack a facile therapeutic approach aimed at correcting alterations in Ca2+ and SERCA function in the heart. However, modulation of SERCA activity through manipulation of PLN or small peptides like it represents a novel and straightforward therapeutic approach. Our lab has identified a novel micropeptide encoded by a muscle-specific RNA currently annotated as a long non-coding RNA (lncRNA), which we have named DWORF (DWarf Open Reading Frame). DWORF shares strong sequence homology and predicted domain structure with the well-described SERCA inhibitors PLN and SLN and our preliminary results show it localizes to the SR and can bind directly to SERCA. We hypothesize that DWORF is functionally homologous to PLN and SLN and that it regulates SERCA activity and myocyte contractility. We will utilize biochemical, cell-based, and genetic approaches, along with in vivo physiological studies to define the function and regulation of DWORF in the heart and to examine how it might impact cardiac disease susceptibility. Collectively, our studies will provide new insights into Ca2+ cycling and regulation in the heart and potentially provide the basis for development of a novel therapeutic target in cardiovascular disease.

描述（由适用提供）：心血管疾病（CVD）是工业化国家的死亡和残疾的主要原因，并且其流行率在发展中国家迅速上升。特别是，心力衰竭（HF）被列为流行病，是与大量发病率，死亡率和医疗保健相关的惊人临床和公共卫生问题支出。当前的疗法仅延迟慢性收缩性心力衰竭患者的发病率和死亡率，因为疾病进展通常会继续延长导致死亡。迫切需要进行心力衰竭的新型疗法，这将需要对这种疾病的基本分子机制有更深入的了解。心脏失败的心脏的普遍特征受到CA2+通过肌浆网（SR）的循环损害，实际上，主要的心肌细胞始终显示出缺陷的兴奋性合同耦合，其特征是SR CA2+序列的减小和延迟的较小的固定型及其静脉内的CAC2+静脉内的抑制作用，而抑制了抑制作用的抑制作用，这些抑制作用的抑制作用是造成不足的抑制作用，这些抑制作用造成了不足的效果。心。 Ca2+将重新隔离为SR，由Ca2+ -ATPase（SERCA）介导，其活性被小型整合性膜蛋白磷脂（PLN）（PLN）和Sarcolipin（SLN）逆转调节。通过将SR加载更高的Ca2+水平来增加Ca2+释放，从而增强SERCA活性和功能是治疗HF的重要临床方法，从而在收缩期间产生更大的肌细胞收缩力。除了基因疗法实际上取代SERCA蛋白（目前在人类临床踪迹中具有AAV载体）外，我们目前缺乏一种旨在纠正CA2+和SERCA功能变化的便利理论方法。然而，通过操纵PLN或小胡椒的调制，它代表了一种新颖而直接的治疗方法。我们的实验室已经确定了一种新型的微肽，该菌皮由当前注释为长的非编码RNA（LNCRNA）的肌肉特异性RNA编码，我们将其命名为Dworf（矮人开放阅读框）。 Dworf与良好描述的SERCA抑制剂PLN和SLN共享强序同源性和预测域结构，我们的初步结果表明它本地化与SR并可以直接结合到SERCA。我们假设Dworf在功能上与PLN和SLN同源，并且它调节SERCA活性和心肌收缩力。我们将利用生化，基于细胞的和遗传学方法以及体内生理研究来定义Dworf在心脏中的功能和调节，并研究它可能影响心脏病敏感性。总的来说，我们的研究将为心脏中的CA2+循环和调节提供新的见解，并有可能为发展中新型治疗靶点的基础提供了心血管疾病的基础。