Dynamic changes of the Nav1.5 interactome and contributions to heart failure

Nav1.5 相互作用组的动态变化及其对心力衰竭的影响

基本信息

批准号：
10478131
负责人：
Steven O Marx
金额：
$ 67.97万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-01 至 2025-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10478131
关键词：
ANK3 gene Ablation Action Potentials Affect Anterior Arrhythmia Arteries Binding Biotin Brugada syndrome Cardiac Cardiac Myocytes Cardiomyopathies Chronic Complex Congestive Heart Failure Coupled Development Dilated Cardiomyopathy Disease Fibroblast Growth Factor Functional disorder Generations Goals Heart failure In Situ Individual Infusion procedures Inherited Intercalated disc Investigation Ion Channel Isoproterenol Knock-out Label Lateral Left Life Ligase Ligation Long QT Syndrome Macromolecular Complexes Maps Mass Spectrum Analysis Membrane Modeling Multiprotein Complexes Mus Mutation Neighborhoods Pathologic Peroxidases Phosphoproteins Physiological Property Proteins Proteomics Records Regulation Risk Role Signal Transduction Sodium Channel Specificity Validation ascorbate candidate validation cohort constriction defined contribution design fibroblast growth factor 13 innovation insight junctophilin knock-down member mouse model mutant novel novel strategies overexpression pressure protective effect sodium channel proteins tool voltage

项目摘要

The NaV1.5 voltage-gated Na+ channel encoded by SCN5A is the fundamental component of macromolecular protein complexes that initiate the cardiac action potential. Abnormal NaV1.5 function is a prominent substrate for inherited and acquired forms of cardiac arrhythmias, reflected by a staggering array of identified NaV1.5 mutations. A small subset of these are associated with dilated cardiomyopathy but the underlying mechanisms are not known. A leading hypothesis, that the arrhythmias drive the cardiomyopathy, cannot explain why most arrhythmogenic NaV1.5 mutations do not cause cardiomyopathy nor why knockout of the NaV1.5 interacting protein FGF13 leads to arrhythmias yet is protective for pressure overload-induced heart failure (HF) despite associated NaV1.5 dysfunction. Moreover, HF from other causes leads to pathological remodeling that disrupts regulation of the VGSC macromolecular complex and increases arrhythmia risk through mechanisms that are poorly understood. Complicating mechanistic insight is that there are different NaV1.5 pools defined by distinct subcellular localizations with the cardiomyocyte, each hypothesized to have protein partners that uniquely define the distinct pools and confer specific channel properties and functions. However, the critical partners remain poorly understood because of challenges of low throughput “favorite” candidate approaches. We propose an unbiased multilevel discovery strategy, employing newly developed second generation proximity labeling tools, novel mouse models, coupled with carefully calibrated cross comparisons designed to increase the specificity of our findings. Exploiting the expertise from two labs with individual and collaborative track records applying a large tool set to dissect complex physiologic mechanisms and define perturbations in pathological states, we propose adaptable candidate validation approaches to establish a comprehensive picture of NaV1.5 interactomes under physiological states and when perturbed by disease. With these innovative approaches we propose to: 1) Define the static and dynamic NaV1.5 channel interactomes and “neighborhoods” within distinct subcellular pools; 2) Elucidate how HF alters the NaV1.5 microenvironment; and 3) Determine the HF-protective effects for ablation of the NaV1.5 interactor, FGF13. With these aims, our goals are to define the contributions of the NaV1.5 macromolecular to development and progression of HF and its associated arrhythmias and to unravel how HF perturbs the NaV1.5 complex to increase arrhythmia risk and exacerbate HF in a vicious cycle.

由 SCN5A 编码的 NaV1.5 电压门控 Na+ 通道是启动心脏动作电位的大分子蛋白复合物是 NaV1.5 功能异常。遗传性和获得性心律失常的一个重要基础，反映在令人震惊的一系列已确定的 NaV1.5 突变，其中一小部分与扩张型心肌病相关。但潜在的机制尚不清楚。一个主要的假设是心律失常驱动。心肌病，无法解释为什么大多数致心律失常 NaV1.5 突变不会引起心肌病以及为什么敲除 NaV1.5 蛋白 FGF13 会导致心律失常尽管存在 NaV1.5 功能障碍，但对压力超负荷引起的心力衰竭 (HF) 具有保护作用。此外，其他原因引起的心力衰竭会导致病理性重塑，从而扰乱 VGSC 的调节大分子复合物，并通过人们知之甚少的机制增加心律失常风险。复杂的机制洞察是，不同的亚细胞定义了不同的 NaV1.5 库。与心肌细胞的定位，每个都发展为具有独特定义的蛋白质伙伴不同的池和特定的渠道属性和功能然而，关键的合作伙伴仍然存在。由于低吞吐量“最喜欢的”候选方法的挑战，人们对此知之甚少。我们提出了一种公正的多层次发现策略，采用新开发的第二层一代邻近标记工具、新颖的小鼠模型，加上仔细校准的交叉旨在提高我们研究结果的特异性的比较，利用两个实验室的专业知识。具有个人和协作跟踪记录，应用大型工具集来剖析复杂的生理学机制并定义病理状态的扰动，我们提出自适应候选验证建立生理状态下 NaV1.5 相互作用组的全面图景的方法当受到疾病困扰时，我们建议：1）定义静态和静态。不同亚细胞池内的动态 NaV1.5 通道相互作用体和“邻域”2) 阐明； HF 如何改变 NaV1.5 微环境；3) 确定 HF 对消融的保护作用 NaV1.5 相互作用因子 FGF13。带着这些目标，我们的目标是确定 NaV1.5 大分子对心力衰竭及其相关心律失常的发生和进展，并揭示心力衰竭如何扰乱心律失常 NaV1.5复合物会增加心律失常风险并加剧心力衰竭，形成恶性循环。