Dissecting the role of novel calcium regulatory proteins in the cardiovascular system

剖析新型钙调节蛋白在心血管系统中的作用

• 批准号: 9505666
• 负责人: Catherine A Makarewich
• 金额: $ 8.84万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-05-01 至 2020-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9505666
• 关键词: Animals; Apoptosis; Award; Biological; Blood Vessels; Ca(2+)-Transporting ATPase; Calcium; Cardiac; Cardiac Myocytes; Cardiovascular Diseases; Cardiovascular Models; Cardiovascular system; Cause of Death; Cell physiology; Characteristics; Companions; Complex; Developed Countries; Developing Countries; Development; Diet; Dilated Cardiomyopathy; Disease; Down-Regulation; Dwarfism; Endoplasmic Reticulum; Endothelial Cells; Energy Metabolism; Etiology; Event; Family; Foundations; Functional disorder; Gene Combinations; Gene Delivery; Genes; Genetic; Health; Heart; Heart failure; High Fat Diet; Homeostasis; Impairment; In Vitro; Ions; Knockout Mice; Knowledge; Malignant Neoplasms; Mediating; Mentors; Metabolic Diseases; Metabolic dysfunction; Metabolic syndrome; Metabolism; Modeling; Molecular; Mus; Muscle Cells; Muscle Contraction; Nodal; Non-Insulin-Dependent Diabetes Mellitus; Obese Mice; Obesity; Open Reading Frames; Pathway interactions; Patients; Peptides; Phase; Phenotype; Physiological; Physiological Processes; Play; Prevalence; Procedures; Proteins; Pump; Regulation; Research; Resistance; Reticulum; Role; Science; Scientist; Second Messenger Systems; Site; Smooth Muscle Myocytes; Stress; Supervision; Techniques; Testing; Therapeutic; Time; Tissues; Training; Transgenes; cardiovascular disorder therapy; career; cell type; design; disability; effective therapy; experience; genetic regulatory protein; heart function; in vivo; inhibitor/antagonist; insight; loss of function; mouse model; novel; novel therapeutics; overexpression; phospholamban; programs; sarcolipin; sarcoplasmic reticulum calcium ATPase; skills; success; symptom treatment; treatment strategy

Project Summary Cardiovascular disease (CVD) is the leading cause of death in the world. Current therapies for CVD aim to treat the symptoms rather than the underlying cause of the disease, thus we desperately need novel insights into the molecular mechanisms of CVD to develop new therapies. Calcium (Ca2+) ions play critical roles in fundamental physiological processes in the cardiovascular system that underlie tissue homeostasis and pump function. A universal characteristic of the failing heart is impaired Ca2+ cycling through the sarco/endoplasmic reticulum (S/ER), the major internal store of Ca2+ in cardiomyocytes. Ca2+ re-sequestration into the S/ER is mediated by a Ca2+-ATPase pump (SERCA), whose activity is regulated by phospholamban (PLN) and sarcolipin (SLN). In the past 3 years, we have identified 4 additional conserved small proteins that regulate SERCA activity in myocytes and non-myocyte cell types, consisting of myoregulin (MLN), endoregulin (ELN), anotheregulin (ALN) and dwarf open reading frame (DWORF). This proposal outlines a comprehensive plan to dissect the function of these peptides to provide critical insights into the complex mechanism of SERCA regulation and Ca2+ homeostasis. To accomplish this, I will employ the following 3 specific aims: 1. To examine the role of ELN and ALN as nodal regulators of SERCA activity, 2. To analyze the in vivo function of the cardiac expressed SERCA-regulins in health and disease, and 3. To investigate the role of Ca2+ dysregulation and SERCA dysfunction in the development of metabolic disorders and cardiovascular disease. The K99 portion of this proposal will be carried out in the lab of the renowned molecular biologist, Dr. Eric Olson. Under Dr. Olson’s guidance, I will functionally characterize the phenotype of ALN and ELN loss-of-function mice and elucidate their mechanism of action (Aim 1). Aims 2a and 2b, which are dedicated to characterizing the cardiomyocyte expressed peptides (ALN, PLN and DWORF), will be initiated under Dr. Olson’s supervision and continued during the independent phase. Finally, the techniques to evaluate systemic metabolism detailed in Aim 3 represent new research experiences for me and will require extensive training during the K99 phase of my award. The knowledge and skills that I acquire during this time will serve as critical components of the foundation of my own independent lab. In summary, the comprehensive plan proposed in this Pathway to Independence Award application will provide me with the opportunity to continue to develop an independent research program that will be instrumental in my success as a scientist. Importantly, the collective body of work generated through the completion of the proposed Aims will make major contributions to the field of cardiovascular science and will set the tone for my career.

项目概要 心血管疾病（CVD）是世界上主要的死亡原因。目前的 CVD 治疗旨在治疗 疾病的症状而不是根本原因，因此我们迫切需要对疾病有新的见解 CVD的分子机制，以开发新疗法。钙 (Ca2+) 离子在基本功能中发挥着关键作用 心血管系统中构成组织稳态和泵功能的生理过程。一个 衰竭心脏的普遍特征是通过肌浆/内质网的 Ca2+ 循环受损 (S/ER)，心肌细胞内 Ca2+ 的主要内部储存。 Ca2+ 重新封存在 S/ER 中是由 Ca2+-ATPase 泵 (SERCA)，其活性受磷蛋白 (PLN) 和肌磷脂 (SLN) 调节。在 过去 3 年，我们鉴定了另外 4 个保守的小蛋白，可调节心肌细胞中的 SERCA 活性 和非肌细胞类型，包括肌调节蛋白 (MLN)、内皮调节蛋白 (ELN)、另一种调节蛋白 (ALN) 和矮细胞 开放阅读框（DWORF）。该提案概述了一个全面的计划来剖析这些功能 肽为 SERCA 调节和 Ca2+ 稳态的复杂机制提供重要见解。 为了实现这一目标，我将采用以下 3 个具体目标： 1. 研究 ELN 和 ALN 作为节点的作用 SERCA活性的调节因子，2.分析心脏表达的SERCA调节蛋白的体内功能 健康和疾病，以及 3. 研究 Ca2+ 失调和 SERCA 功能障碍在 代谢紊乱和心血管疾病的发展。该提案的K99部分将被采纳 在著名分子生物学家埃里克·奥尔森 (Eric Olson) 博士的实验室中。在奥尔森博士的指导下，我将在功能上 表征 ALN 和 ELN 功能丧失小鼠的表型并阐明其作用机制（Aim 1）。目标 2a 和 2b，致力于表征心肌细胞表达的肽（ALN、PLN 和 DWORF），将在 Olson 博士的监督下启动，并在独立阶段继续进行。 最后，目标 3 中详述的评估全身代谢的技术代表了新的研究经验 对我来说，在我的奖励的 K99 阶段需要进行广泛的培训。我所掌握的知识和技能 在此期间获得的成果将成为我自己的独立实验室基础的关键组成部分。 总之，独立之路奖申请中提出的全面计划将提供 我有机会继续开发一个独立的研究项目，这将有助于我 作为一名科学家的成功。重要的是，通过完成拟议的工作而产生的集体工作 目标将为心血管科学领域做出重大贡献，并将为我的职业生涯定下基调。