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Endothelial Lineage Diversity: Role of Epigenetics

内皮谱系多样性：表观遗传学的作用

基本信息

批准号：
8001269
负责人：
William C Aird
金额：
$ 27.07万
依托单位：
BETH ISRAEL DEACONESS MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2010
资助国家：
美国
起止时间：
2010-08-01 至 2015-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8001269
关键词：
Adult Area Attention Blood Circulation Blood Vessels Blood flow Cardiovascular system Cell Differentiation process Cell Lineage Cell Maintenance Cell model Cell physiology Cells Chromatin Clinical Complex Custom DNA DNA Methylation DNA Sequence Development Diagnosis Disease Elements Embryo Endothelial Cells Epigenetic Process Event Functional RNA Future Gene Expression Gene Mutation Genes Genetic Transcription Genomics Goals Health Heterogeneity Histone Code Hypoxia Immunoprecipitation In Situ In Vitro Laboratories Mediating Modification Molecular Mus Mutation Nucleic Acid Regulatory Sequences Nucleosomes Pathway interactions Pattern Phenotype Play Polycomb Process Production Program Research Project Grants Regenerative Medicine Regulation Regulatory Pathway Reporting Resolution Role Site Smooth Muscle Myocytes Stimulus Time Transcriptional Activation Vascular Endothelium Work base cDNA Arrays cell type defined contribution density embryonic stem cell gene repression histone modification human NOS3 protein human disease in vitro Model in vivo insight interest laser capture microdissection member pluripotency promoter research study sodium bisulfite stem cell differentiation success

项目摘要

The scientific goal of this project is to gain further insjght into how chromatin-based pathways contribute to endothelial cell (EC) gene expression. A focus of our studies is the gene responsible for the production of NO by vascular endothelium, namely endothelial nitric oxide synthase (eNOS). We have reported that the promoter of the eNOS gene is hypomethylated in endothelial cells both in vitro and in vivo. In contrast, the promoter was densely methylated in genomic DNA isolated from cell types that do not express eNOS. We have also reported that the nucleosomes that encompass the eNOS core promoter were highly enriched in activating histone modifications in expressing versus non-expressing cell types. The overall hypothesis of this project is that epigenetic processes play a major role in the spatial and temporal regulation of EC gene expression. This project is extremely complementary to our overall PPG theme. We will collaborate with all members of this PPG and take full advantage of the PPG Cores. In Aim I we will define whether the maintenance of cell-specific epigenetic marks at the eNOS proximal promoter is critically important for maintaining constitutive transcription of eNOS in ECs and transcriptional repression in differentiated cell types that do not express eNOS. In Aim II we will define the contribution of epigenetic pathways to the temporal regulation of eNOS expression during EC differentiation and vascular development. We will use in vitro and in vivo approaches. We have found differential patterns of epigenetic modifications at the promoters of a number of EC-specific genes suggesting that our findings with the eNOS gene may be broadly relevant. Therefore, in Aim III we will use a ChlP-on-chip approach to define the contribution of epigenetic pathways to the global regulation of EC-specific gene expression. We will also identify large-intergenic non-coding RNAs that especially enriched in ECs. We anticipate that our studies will provide new insight into the contribution of epigenetic pathways to global patterns of EC-specific gene expression

该项目的科学目标是进一步深入了解基于染色质的途径如何促进内皮细胞（EC）基因表达。我们的研究重点是负责血管内皮产生一氧化氮的基因，即内皮型一氧化氮合酶（eNOS）。我们已经报道了在体外和体内内皮细胞中eNOS基因的启动子是低甲基化的。相反，从不表达eNOS的细胞类型中分离的基因组DNA中，启动子被密集甲基化。我们还报道了包含eNOS核心启动子的核小体在表达与非表达细胞类型中激活组蛋白修饰时高度富集。本项目的总体假设是表观遗传过程在EC基因表达的时空调控中起主要作用。这个项目是对我们整体PPG主题的极大补充。我们将与PPG的所有成员合作，充分利用PPG的核心。在Aim I中，我们将确定eNOS近端启动子上细胞特异性表观遗传标记的维持是否对维持ec中eNOS的组成性转录和不表达eNOS的分化细胞类型的转录抑制至关重要。在Aim II中，我们将定义表观遗传途径对EC分化和血管发育过程中eNOS表达的时间调控的贡献。我们将使用体外和体内方法。我们在许多ec特异性基因的启动子上发现了不同的表观遗传修饰模式，这表明我们对eNOS基因的研究结果可能具有广泛的相关性。因此，在Aim III中，我们将使用芯片上的chlp方法来定义表观遗传途径对ec特异性基因表达的全球调控的贡献。我们还将鉴定在ECs中特别富集的大基因间非编码rna。我们期望我们的研究将提供新的见解