Desmosomes in cardiomyocyte homeostasis and disease

桥粒在心肌细胞稳态和疾病中的作用

基本信息

批准号：
10606894
负责人：
William Tswenching Pu
金额：
$ 81.65万
依托单位：
BOSTON CHILDREN'S HOSPITAL
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-12-15 至 2026-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10606894
关键词：
ATAC-seq Ablation Acceleration Adhesives Adipose tissue Adult Arrhythmia Biomedical Engineering Cardiac Cardiac Myocytes Cardiomyopathies Cell Nucleus Cells Chemicals Complex DISC components Data Desmosomes Development Disease Dissection Elements Experimental Models Fluorescent in Situ Hybridization Functional disorder Gene Expression Gene Expression Regulation Genes Genetic Genetic Transcription Goals Heart Abnormalities Heart failure Homeostasis Human In Situ Hybridization Intercalated disc Knowledge Lead Life Link Metabolism Methods Modeling Molecular Mus Muscle function Mutation Myocardial Contraction Myocardial dysfunction Myocardium N-Cadherin Pathogenesis Pathway interactions Patients Periodicity Phenotype Plasma Proteins Proteomics Regulatory Element Relaxation Resolution Role Sarcomeres Signal Transduction Testing Ventricular Arrhythmia WNT Signaling Pathway arrhythmogenic cardiomyopathy candidate identification candidate selection defined contribution desmoplakin follow-up gain of function genetic approach heart rhythm improved in vivo induced pluripotent stem cell inhibitor innovation insight loss of function mosaic multiple omics mutant novel preservation protein function single molecule single nucleus RNA-sequencing transcriptome transcriptomics

ATAC-seq Ablation Acceleration Adhesives Adipose tissue Adult Arrhythmia Biomedical Engineering Cardiac Cardiac Myocytes Cardiomyopathies Cell Nucleus Cells Chemicals Complex DISC components Data Desmosomes Development Disease Dissection Elements Experimental Models Fluorescent in Situ Hybridization Functional disorder Gene Expression Gene Expression Regulation Genes Genetic Genetic Transcription Goals Heart Abnormalities Heart failure Homeostasis Human In Situ Hybridization Intercalated disc Knowledge Lead Life Link Metabolism Methods Modeling Molecular Mus Muscle function Mutation Myocardial Contraction Myocardial dysfunction Myocardium N-Cadherin Pathogenesis Pathway interactions Patients Periodicity Phenotype Plasma Proteins Proteomics Regulatory Element Relaxation Resolution Role Sarcomeres Signal Transduction Testing Ventricular Arrhythmia WNT Signaling Pathway arrhythmogenic cardiomyopathy candidate identification candidate selection defined contribution desmoplakin follow-up gain of function genetic approach heart rhythm improved in vivo induced pluripotent stem cell inhibitor innovation insight loss of function mosaic multiple omics mutant novel preservation protein function single molecule single nucleus RNA-sequencing transcriptome transcriptomics

展开

项目摘要

SUMMARY Intercalated disks (ICDs) connect the termini of adjacent cardiomyocytes (CMs) physically, electrically, and chemically. The structural role of ICDs to preserve CM integrity in the face of billions of cycles of forceful con- traction and relaxation is well appreciated; however, the function of ICDs as essential CM signaling hubs is only now emerging. Arrhythmogenic cardiomyopathy (ACM) provides a unique window into the function of ICDs and specifically desmosomes. ACM is a potentially lethal disorder characterized by high arrhythmia bur- den, loss of contractile myocardium, and replacement by fibro-fatty tissue. Mutations of desmosome genes (PKP2, DSG2, DSC2, DSP, JUP) occur in approximately half of ACM patients. Despite growing knowledge about ACM disease pathogenesis, the mechanistic links between desmosome mutations and arrhythmias, my- ocardial dysfunction, and fibrofatty replacement remain poorly understood. The overall goal of this proposal is to gain insights into the mechanisms by which desmosome mutations cause arrhythmia and myocardial dysfunction; Our overarching hypothesis is that desmosomes are inte- gral for maintaining normal cardiomyocyte homeostasis through both their structural and signaling activities. ACM mutations disrupt these activities to cause both loss of structural integrity and aberrant signaling. We will test these hypotheses through four parallel but complementary Specific Aims: (1) We will examine cell composition and gene regulation of human ACM myocardium, using concurrent single nucleus RNA-seq and ATAC-seq, and spatial transcriptomics (snMulti-seq) with massively parallel single molecule fluo- rescent in situ hybridization (MERFISH); (2) We will use mosaic, adult, cardiomyocyte specific inactivation of Dsp to probe the cell autonomous functions of desmosomes. This model will be studied using snMulti-seq and MERFISH, followed by interrogation of key predicted regulators using in vivo gain- and loss-of-function ap- proaches; (3) Using proximity proteomics of ICD component N-cadherin, we identified novel ICD components and ICD components that are altered by Dsp ablation. We will use in vivo gain- and loss-of-function ap- proaches to study the function of selected candidates identified by this screen; (4) Define the contributions of WNT and GSK3 signaling to ACM phenotypes in DSP mutant hiPSC-CMs. Using genetic approaches in bioen- gineered hiPSC-CMs, we will dissect the involvement of GSK3 and WNT signaling to ACM pathogenesis. Impact: This proposal will advance our understanding of the function of desmosomes and ICDs in CM homeostasis and the molecular pathogenesis of ACM. This knowledge will accelerate efforts to develop targeted ACM therapies.

概括插入式磁盘（ICD）连接相邻心肌细胞（CMS）的末端化学。 ICD在数十亿个有力的周期中保持CM完整性的结构作用牵引和放松得到很好的赞赏；但是，ICD作为必需CM信号中心的功能是直到现在出现。心律失常性心肌病（ACM）提供了一个独特的窗口 ICD，特别是脱染色体。 ACM是一种潜在的致命疾病，其特征是高心律失常 DEN，收缩心肌的损失以及纤维脂肪组织的替代。腔体基因的突变（PKP2，DSG2，DSC2，DSP，JUP）发生在大约一半的ACM患者中。尽管知识越来越大关于ACM疾病发病机理，脱骨突变与心律不齐之间的机械联系，my- 心脏功能障碍和纤维状置换术仍然知之甚少。该提案的总体目标是了解脱骨突变的机制导致心律不齐和心肌功能障碍；我们的总体假设是脱糖体是inte的通过其结构和信号传导维持正常心肌稳态的gral 活动。 ACM突变破坏了这些活动，以引起结构完整性的丧失和异常信号。我们将通过四个平行但互补的特定目的来检验这些假设：（1）我们将使用并发的单核检查人ACM心肌的细胞组成和基因调节 RNA-SEQ和ATAC-SEQ，以及空间转录组学（SNMULTI-SEQ），具有大量平行的单分子氟恢复原位杂交（Merfish）；（2）我们将使用马赛克，成人，心肌细胞特异性灭活 DSP探测脱糖体的细胞自主函数。该模型将使用snmulti-seq和 Merfish，然后使用体内获得和功能丧失的关键预测调节剂进行审问促进；（3）使用ICD成分N-钙粘蛋白的接近蛋白质组学，我们确定了新型ICD成分以及通过DSP消融改变的ICD组件。我们将使用体内增益和功能丧失ap- 促进该屏幕确定的选定候选者的功能；（4）定义的贡献 Wnt和GSK3信号传导DSP突变HIPSC-CM中的ACM表型。在生物元中使用遗传方法 Gineered HipsC-CMS，我们将剖析GSK3和Wnt信号传导与ACM发病机理的参与。影响：该提案将促进我们对CM中脱粒和ICD的功能的理解稳态和ACM的分子发病机理。这些知识将加速发展的努力靶向ACM疗法。