Intercalated disc-nuclear lamina coupling as a molecular substrate for arrhythmogenic cardiomyopathy

闰盘-核层耦合作为致心律失常性心肌病的分子底物

基本信息

批准号：
10713689
负责人：
Eric M Small
金额：
$ 69.92万
依托单位：
UNIVERSITY OF ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-08-01 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10713689
关键词：
Acceleration Adhesions Architecture Arrhythmia Arrhythmogenic Right Ventricular Dysplasia Attention Cardiac Cardiac Myocytes Cardiomyopathies Cell Communication Cell Death Cell Nucleus Cell membrane Cell surface Cell-Matrix Junction Cells Chromatin Code Complex Coupling Cytoskeleton DNA Damage Data Desmin Desmosomes Disease Disease Progression Ecosystem Filament Foundations Functional disorder Gene Expression Genes Genetic Diseases Genetic Epistasis Genetic Transcription Genome Genomics Heart Arrest Hereditary Disease Homeostasis Human Image Induced Mutation Intercalated disc Intermediate Filaments Knowledge Lamin Type A Lamins Life Link Mechanics Mediating Methods Microscopy Microtubules Modernization Molecular Morbidity - disease rate Mus Muscle Cells Mutant Strains Mice Mutation Myocardial Myocardial dysfunction Myocardium Nuclear Nuclear Envelope Nuclear Lamina Nuclear Structure Onset of illness Organelles Pathogenesis Pathologic Pathway interactions Patients Physiology Proteins Proteome Proteomics Publishing Reporting Research Role Sampling Signal Transduction Structure Surface TP53 gene Testing Tissues Translating Variant Ventricular Ventricular Arrhythmia Work arrhythmogenic cardiomyopathy desmoplakin disease phenotype druggable target env Gene Products experimental study gene repression mechanical load molecular imaging mortality mouse model novel therapeutic intervention pharmacologic plakophilin 2 prevent response scaffold single molecule transcriptional reprogramming transcriptomics transmission process

项目摘要

Plakophilin-2 (PKP2) is classically defined as a protein of the desmosome, an intercellular adhesion structure residing in the cardiac intercalated disc (ID). Mutations in PKP2 associate with most cases of gene-positive arrhythmogenic right ventricular cardiomyopathy (ARVC), a disease characterized by high propensity to life- threatening arrhythmias and myocardial structural damage, often of right ventricular predominance. Much attention has been given to the loss of cell-cell attachment at the ID as a disease mechanism. Yet, it is becoming evident that PKP2 mutations also lead to an array of poorly understood cardiomyocyte (CM)-intrinsic disturbances. Desmin intermediate filaments are anchored to the desmosome in a PKP2-dependent manner, supporting CM structural integrity and facilitating communication from the cell surface to the nucleus. Our prior work in mouse models and human patient samples found PKP2 mutation disrupts CM nuclear envelope (NE) integrity and leads to DNA damage. Based on published reports and our preliminary data, we hypothesize that PKP2 deficiency, or disease relevant PKP2 mutations, disrupt the structural, functional and molecular integrity of the cardiomyocyte nuclear envelope, leading to genomic reorganization, the DNA damage response, and altered transcription. The following aims will investigate how PKP2 deficiency disrupts the nucleus to accelerate ARVC disease progression. Aim 1: Define the impact of PKP2 deficiency on the cardiomyocyte nuclear lamina protein interactome. We hypothesize that PKP2 deficiency alters the proteome of the cardiomyocyte NE, and that this disruption is an early trigger for the disease phenotype. We will interrogate changes in the molecular ecosystem of the cardiomyocyte NE after loss of PKP2 expression using proteomics and single molecule imaging. Aim 2: Define the impact of PKP2 deficiency on cardiomyocyte genomic organization. We hypothesize that loss of NE integrity in PKP2 deficient CMs disrupts genomic organization at Lamin Associated Domains and causes transcriptional remodeling. We will determine how structural damage is transmitted from the cell membrane to the genome, focusing on changes that occur in the vicinity of the NE through advanced imaging, genomic and transcriptomic approaches. Aim 3: Investigate strategies to reduce DDR and delay cardiomyopathy in PKP2 deficient mice. We hypothesize data that PKP2 mutation induces P53-dependent DNA damage response (DDR), which may exacerbate ARVC disease progression. Genetic epistasis experiments and pharmacological approaches will investigate how the P53-dependent DDR contributes to PKP2-dependent cardiomyopathy. Defining pathological changes to nuclear architecture that precede overt myocardial structural remodeling will reveal exciting opportunities for new therapeutic strategies aimed at slowing ARVC disease progression by restoring nuclear envelope homeostasis or preventing the DNA damage response.

plakophilin-2（PKP2）经典定义为脱粒的蛋白质，一种细胞间粘附结构居住在心脏插入的椎间盘（ID）中。 PKP2中的突变与大多数基因阳性情况相关心律失常右心肌病（ARVC），这种疾病以高倾向的特征威胁性心律不齐和心肌结构损害，通常是右心室优势。很多注意将ID处于疾病机制的细胞细胞附着的丧失引起人们的注意。然而，它正在变成显然，PKP2突变也导致一系列知识不足的心肌细胞（CM） - intrinsic 干扰。 Desmin中间丝以PKP2依赖性方式锚定在脱骨体上，支持CM结构完整性并促进从细胞表面到核的通信。我们的先验在小鼠模型和人类患者样品中的工作发现PKP2突变破坏了CM核包膜（NE）完整性并导致DNA损伤。根据已发表的报告和我们的初步数据，我们假设 PKP2缺乏症或疾病相关的PKP2突变破坏了结构，功能和分子完整性心肌细胞核包膜，导致基因组重组，DNA损伤反应和转录改变。以下目的将研究PKP2缺乏如何破坏核以加速 ARVC疾病进展。目标1：定义PKP2缺乏对心肌细胞核薄片蛋白相互作用组的影响。我们假设PKP2缺乏改变了心肌细胞的蛋白质组，并且这种干扰是疾病表型的早期触发因素。我们将询问对分子生态系统的变化使用蛋白质组学和单分子成像失去PKP2表达后的心肌细胞。目标2：定义PKP2缺乏对心肌细胞基因组组织的影响。我们假设PKP2缺陷CMS中NE完整性的丧失会破坏层固定的基因组组织相关域并引起转录重塑。我们将确定结构性损害从细胞膜传播到基因组，重点是在NE附近发生的变化通过先进的成像，基因组和转录组方法。 AIM 3：研究减少DDR和延迟心肌病的策略。我们假设数据表明PKP2突变诱导p53依赖性DNA损伤响应（DDR），这可能恶化的ARVC疾病进展。遗传上的实验和药理学方法将研究依赖p53的DDR有助于PKP2依赖性心肌病。定义对明显心肌结构重塑之前的核结构的病理变化将为新的治疗策略揭示激动人心的机会，旨在通过恢复核包膜稳态或防止DNA损伤反应。