Dynamics of cardiac nuclei in heart disease

心脏病中心肌细胞核的动力学

• 批准号: 10523028
• 负责人: Thomas M. Vondriska
• 金额: $ 39万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-01-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10523028
• 关键词: ATAC-seq; Adult; Architecture; Biology; Cardiac; Cardiac development; Cardiovascular system; Cell Nucleus; Cells; Chromatin; Chromatin Loop; Chromatin Modeling; Chromatin Structure; Data; Development; Disease; Disease Progression; EP300 gene; Engineering; Enterobacteria phage P1 Cre recombinase; Environment; Enzymes; Epigenetic Process; Family; Female; Fibroblasts; Fluorescent in Situ Hybridization; Gel; Gene Activation; Gene Expression; Genes; Genetic Materials; Genetic Transcription; Genome; Genomics; Grant; Guide RNA; Heart; Heart Diseases; Heart Hypertrophy; Heart failure; Histology; Histone H1; Histones; Hypertrophy; Incidence; Injury; Interphase Cell; Investigation; Isoproterenol; Job Description; Measures; Microscopy; Mitotic; Modification; Molecular; Molecular Conformation; Molecular Target; Mus; Muscle Cells; Myocardial Infarction; Myofibroblast; Neighborhoods; Nuclear; Organ; Organism; Pathogenesis; Pathologic; Phenotype; Physical condensation; Process; Protein Isoforms; Resolution; Role; Sex Differences; Signal Transduction; Smooth Muscle Actin Staining Method; Structure; Symptoms; Taxes; Testing; Tissues; Transforming Growth Factor beta; Transgenic Mice; Validation; Work; adrenergic stress; base; cell type; chromatin remodeling; experimental study; extracellular; gene repression; genomic locus; heart function; histone modification; in vivo; insight; ischemic injury; loss of function; male; novel; periostin; preservation; pressure; promoter; protein expression; reconstruction; response; response to injury; sex; simulation; single-cell RNA sequencing; three-dimensional modeling; tool; transcriptome; transcriptome sequencing; vector

PROJECT SUMMARY/ABSTRACT Epigenetic processes have been implicated in control of cardiac development and in the incidence and progression of disease. Heart failure in particular has been shown to involve the actions of chromatin remodeling enzymes and to proceed by temporal reorganization of histone modifications and gene expression. The scientific premise of this application is that understanding the principles of chromatin structure-function, including the roles of specific molecular targets such as the linker histone H1 family, is key to re-engineering healthy transcriptomes in the setting of disease. Based on preliminary data implicating its role in fibroblast phenotype, we will mechanistically investigate the role of linker histone H1.0 in chromatin organization, using gain- and loss-of- function approaches in cells and in vivo. We hypothesize that precise structural orientation of the genome is underpinned by cell type-specific molecular processes and that disease results from reorganization of global genome architecture, thereby enabling pathologic gene expression. To test this hypothesis, we will perform the first ever reconstruction of genome topology on distinct cell types—fibroblasts and myocytes—from the same organ. We will precisely measure differences in chromatin architecture in fibroblast nuclei from male and female mice, examining the role of sex differences in genome organization as a potential unexplored contributor to differences in gene expression and cardiovascular phenotype between the sexes. We will use dCas CLOuD9- based chromatin loop reorganization tools to definitively test the causative role of chromatin interactions in transcription and fibroblast activation. Based on preliminary data implicating a privileged role for linker histone H1.0 in cardiac fibroblasts, we will examine the molecular mechanisms whereby H1.0 controls fibroblast gene expression and locus specific chromatin accessibility. We will use gain- and loss-of-function approaches in isolated fibroblasts to examine the role of histone H1.0 to regulate nuclear condensation, fibroblast gel contraction, proliferation and myofibroblast protein expression. We will conclusively determine the in vivo role of histone H1.0 in cardiac fibroblast phenotype under basal conditions and in the setting of pressure overload, beta adrenergic stress by isoproterenol, or ischemic injury. This approach will allow us to test the role of histone H1.0 to regulate assembly of specific chromatin neighborhoods identified in our genome structure studies as well as to examine whether these neighborhoods are reorganized in a histone H1.0-dependent manner in vivo concomitant with development of disease. The proposed experiments will provide mechanistic insights into how histone H1.0 contributes to fibroblast activation and cardiac function in vivo, revealing molecular details underpinning epigenetic control of the heart’s response to injury.

项目摘要/摘要 在控制心脏发展和事件中，已经暗示了表观遗传过程 疾病的进展。特别是心力衰竭涉及染色质重塑的作用 酶并通过暂时重组组蛋白修饰和基因表达进行。科学 此应用的前提是了解染色质结构功能的原理，包括角色 特定分子靶标（例如接头组蛋白H1家族）是重新设计健康转录组的关键 在疾病的情况下。基于初步数据隐含其在成纤维细胞表型中的作用，我们将 使用增益和丧失，机械师研究接头组蛋白H1.0在染色质组织中的作用 细胞和体内的功能方法。我们假设基因组的精确结构取向是 受细胞类型特异性分子过程的基础，该疾病是由全球重组引起的 基因组结构，从而实现病理基因表达。为了检验这一假设，我们将执行 在不同的细胞类型（纤维细胞和心肌细胞）上，基因组拓扑的首次重建。 器官。我们将准确测量男性和女性的成纤维细胞核中染色质结构的差异 小鼠，研究性别差异在基因组组织中的作用，作为潜在的意外贡献者 性别之间基因表达和心血管表型的差异。我们将使用DCAS Cloud9- 基于染色质环的重组工具，以确定测试染色质相互作用在 转录和成纤维细胞激活。基于初步数据，暗示了接头组蛋白的特权角色 H1.0在心脏成纤维细胞中，我们将检查H1.0控制成纤维细胞基因的分子机制 表达和基因座特异性染色质可及性。我们将在 分离的成纤维细胞检查组蛋白H1.0在调节核凝结，成纤维细胞凝胶中的作用 收缩，增殖和肌纤维细胞蛋白表达。我们将最终确定体内的作用 Hisstone H1.0在基本条件下和压力超负荷的情况下，心脏成纤维细胞表型 异丙肾上腺素或缺血性损伤的肾上腺应激。这种方法将使我们能够测试组蛋白H1.0的作用 调节在我们的基因组结构研究中确定的特定染色质社区的组装以及 检查这些社区是否以组蛋白H1.0依赖性方式重组 伴随着疾病的发展。拟议的实验将提供有关如何 Hisstone H1.0在体内有助于成纤维细胞激活和心脏功能，揭示了分子细节 对心脏对损伤的反应的表观遗传控制的基础。