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

项目摘要 心血管疾病（CVD）是世界上主要的死亡原因。目前的CVD疗法旨在治疗 症状，而不是疾病的根本原因，因此我们迫切需要新的见解， CVD的分子机制，以开发新的治疗方法。钙离子（Ca 2+）在基础代谢中起着关键作用。 心血管系统中的生理过程是组织稳态和泵功能的基础。一 心力衰竭的普遍特征是通过肌浆网/内质网的Ca 2+循环受损 （S/ER），心肌细胞中Ca 2+的主要内部储存。Ca 2+再螯合到S/ER中是由一个 Ca ~（2+）-ATP酶泵（SERCA），其活性受受磷蛋白（PLN）和肌磷脂（SLN）调节。在 在过去的3年中，我们已经鉴定出4种额外的调节肌细胞SERCA活性的保守小蛋白 和非肌细胞细胞类型，包括肌调节蛋白（MLN）、内皮调节蛋白（ELN）、另一调节蛋白（ALN）和侏儒细胞（dwarf 打开阅读帧（DWORF）。该提案概述了一个全面的计划，以剖析这些功能， 肽提供关键的洞察SERCA调节和Ca 2+稳态的复杂机制。 为了实现这一点，我将采用以下三个具体目标：1。检查ELN和ALN作为节点的作用 SERCA活性的调节剂，2.为了分析心脏表达的SERCA调节蛋白在体内的功能， 健康与疾病，3。探讨钙离子失调和SERCA功能障碍在脑缺血再灌注损伤中的作用。 代谢紊乱和心血管疾病的发展。该提案的K99部分将被执行 在著名分子生物学家埃里克·奥尔森博士的实验室里在奥尔森博士的指导下，我将 描述ALN和ELN功能丧失小鼠的表型并阐明其作用机制（目的 1）。目的2a和2b，其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中详细介绍的评价全身代谢的技术代表了新的研究经验 在我的K99阶段，我需要广泛的培训。我所掌握的知识和技能 在此期间获得的将作为我自己的独立实验室的基础的关键组成部分。 总而言之，独立之路奖申请中提出的综合计划将提供 我有机会继续发展一个独立的研究计划，这将有助于我 作为科学家的成功重要的是，通过完成拟议的 Aims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。

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.