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+通道是启动心脏作用电位的大分子蛋白复合物。异常nav1.5功能是遗传和获得形式的心律不齐形式的突出底物，反映了惊人已识别的NAV1.5突变阵列。其中一小部分与扩张的心肌病有关但是，基本机制尚不清楚。一个主要假设，即心律不齐驱动心肌病，无法解释为什么大多数心律失常NAV1.5突变不会引起心肌病也不是为什么NAV1.5相互作用蛋白FGF13的敲除导致心律不齐的压力超负荷引起的心力衰竭（HF）的保护性NAV1.5功能障碍。此外，来自其他原因的HF导致病理重塑，破坏了VGSC的调节大分子复合物并通过知之甚少的机制增加心律不齐的风险。复杂的机械洞察力是，有不同的NAV1.5池由不同的亚细胞定义与心肌细胞的局部定位，每个人都有蛋白质伴侣的蛋白质伴侣，可以独特地定义不同的池和会议特定的渠道属性和功能。但是，关键伙伴仍然由于低吞吐量“最喜欢的”候选方法的挑战，理解不佳。我们提出了一种公正的多级发现策略，采用新开发的第二生成接近标签工具，新型鼠标模型，再加上精心校准的十字架旨在提高我们发现的特异性的比较。利用两个实验室的专业知识随着个人和协作记录，应用大型工具集来剖析复杂的生理机制和定义病理状态的扰动，我们提出了适应性的候选验证在物理状态下建立NAV1.5相互作用的全面图片的方法当疾病扰动时。通过这些创新的方法，我们建议：1）定义静态和动态NAV1.5通道相互作用和“邻域”在不同的亚细胞池中； 2）阐明 HF如何改变NAV1.5微环境； 3）确定消融的HF保护作用 NAV1.5交互式，FGF13。以这些目的，我们的目标是定义NAV1.5大分子的贡献 HF及其相关心律失常的发展和进展，并揭示HF如何渗透 NAV1.5复合物在恶性循环中增加心律不齐的风险和加剧HF。