The Role of 3-Dimensional Genome Integrity In Cardiac Laminopathies

三维基因组完整性在心脏核纤层蛋白病中的作用

• 批准号: 10417144
• 负责人: Ioannis Karakikes
• 金额: $ 52.64万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10417144
• 关键词: 3-Dimensional; Architecture; Arrhythmia; Biological Assay; Ca(2+)-Calmodulin Dependent Protein Kinase; Calcium; Cardiac; Cardiac Myocytes; Cells; Chromatin; Clustered Regularly Interspaced Short Palindromic Repeats; Coupling; Data; Development; Dilated Cardiomyopathy; Disease; Disease Progression; Distal; Etiology; Functional disorder; Gene Expression; Genes; Genetic Transcription; Genome; Genomics; Goals; Heart failure; Homeobox; Human; Impairment; In Vitro; Knowledge; Lamin Type A; Lead; Length; Life; Link; Mediating; Mediator of activation protein; Modeling; Molecular; Molecular Target; Mutate; Mutation; Nuclear; Nuclear Envelope; Nuclear Lamina; PDGFRB gene; Pathogenesis; Patients; Phenotype; Physiology; Platelet-Derived Growth Factor Receptor; Play; Positioning Attribute; Process; Proteins; Proteome; Proteomics; Research Proposals; Role; Signal Transduction; Small Interfering RNA; Structure; Technology; Therapeutic; Transcriptional Regulation; Translating; Up-Regulation; base; calmodulin-dependent protein kinase II; chromatin protein; computerized tools; design; epigenome; epigenomics; genome integrity; genome-wide; genomic locus; heart cell; human disease; induced pluripotent stem cell; induced pluripotent stem cell derived cardiomyocytes; induced pluripotent stem cell technology; mammalian genome; multidisciplinary; mutant; new technology; novel therapeutics; precision medicine; restoration; sound; stem cell model; stem cells; sudden cardiac death; targeted treatment; transcriptome

Project Summary Mutations in the LMNA gene, which encodes type-A lamin proteins that constitute the nuclear lamina network, cause a broad spectrum of diseases collectively called laminopathies. LMNA is one of the most frequent genes involved in dilated cardiomyopathy (DCM), an incurable disease characterized arrhythmias and sudden cardiac death. How mutations in LMNA cause DCM is poorly understood. In the mammalian genome, lamin A interacts with chromatin in a cell-specific manner at lamina-associated domains (LADs) at the nuclear periphery, suggesting a key role for lamin A proteins in the overall genome integrity and transcriptional regulation. However, where and how lamin A interacts with the genome and whether DCM-causing lamin A mutations rearrange the genome architecture in DCM remains elusive. The central hypothesis of this proposal is that a subset of LMNA mutations cause DCM through loss of structural function of lamin A, leading to spatial reorganization of the genome architecture and aberrant gene expression. This hypothesis is based on strong preliminary data showing that a DCM LMNA mutation causes haploinsufficiency and triggers genome-wide changes in LADs concomitantly with aberrant gene expression in patient-specific human induced pluripotent stem cells derived cardiomyocytes (iPSC-CMs). Together our data suggest a mechanistic link between the dysregulation of genome integrity and the DCM phenotype. The overarching goal of the study is to understand the molecular etiology responsible for the cardiac-specific phenotypes caused by LMNA mutations and to demonstrate that impaired lamin A chromatin- and protein-interactions cause DCM. In Aim 1 we will validate the disruption of the 3D genome reorganization as a causative mechanism of LMNA-related DCM. In Aim 2 we will decipher the molecular basis of calcium dysregulation and arrhythmia in LMNA DCM, and in Aim 3 we will develop a therapeutic strategy by modulating the local genome organization. We provided compelling preliminary data to support the soundness of our hypothesis-driven research proposal. We have assembled a multidisciplinary team and we are well positioned to achieve the project goals within four years. If successful, our studies will provide a new paradigm for understanding the pathogenesis and pave the way for targeted therapies for LMNA-related DCM.

项目摘要 LMNA基因中的突变，该突变编码构成核层网络的A型层粘连蛋白， 引起广泛的疾病，统称为椎板病。 LMNA是最常见的基因之一 参与扩张的心肌病（DCM），一种无法治愈的疾病，表征心律不齐和突然心脏 死亡。 LMNA中的突变导致DCM的理解很少。在哺乳动物基因组中，层粘连a有相互作用 在核周围的层状相关结构域（LAD）处的染色质特异性方式， 提出层固定蛋白在整体基因组完整性和转录调控中的关键作用。 但是，层粘连蛋白A与基因组的相互作用以及如何引起DCM引起层粘连蛋白A突变 重新排列DCM中的基因组建筑仍然难以捉摸。该提议的核心假设是 LMNA突变的子集导致DCM通过层粘连蛋白A的结构功能丧失，从而导致空间 基因组结构和异常基因表达的重组。该假设基于强大 初步数据表明，DCM LMNA突变会导致单倍不足并触发全基因组 在患者特异性人类诱导的多能中，LAD的变化与异常的基因表达发生变化 干细胞得出的心肌细胞（IPSC-CMS）。我们的数据一起提出了 基因组完整性和DCM表型的失调。研究的总体目标是了解 负责LMNA突变引起的心脏特异性表型的分子病因 证明层粘连蛋白A染色质和蛋白质互动受损会导致DCM。在AIM 1中，我们将验证 3D基因组重组的破坏是LMNA相关DCM的致病机制。在目标2中我们 将破译LMNA DCM中钙失调和心律不齐的分子基础，在AIM 3中，我们将 通过调节本地基因组组织制定治疗策略。我们提供了引人注目的 初步数据，以支持我们假设驱动的研究建议的合理性。我们已经组装了 多学科团队和我们在四年内实现项目目标的好处。如果成功， 我们的研究将为理解发病机理的新范式提供新的范式 LMNA相关DCM的疗法。