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。我们研究的重点是负责由血管内皮产生NO的基因，即内皮一氧化氮合酶（ENOS）。我们报道说，eNOS基因的启动子在体外和体内均在内皮细胞中脱甲基化。相比之下，启动子在从未表达eNOS的细胞类型中分离的基因组DNA中被密集甲基化。我们还报道说，包含eNOS核心启动子的核小体在表达和非表达细胞类型的激活组蛋白修饰方面高度富集。该项目的总体假设是表观遗传过程在EC基因表达的空间和时间调节中起主要作用。该项目与我们的整体PPG主题非常互补。我们将与该PPG的所有成员合作，并充分利用PPG核心。在目的中，我们将定义eNOS近端启动子处细胞特异性表观遗传标记的维持对于在不表达eNOS的分化细胞类型中维持ECS中eNOS的本构和转录抑制至关重要。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表达的时间调节的贡献。我们将使用体外和体内方法。我们发现，许多EC特异性基因的启动子的表观遗传修饰的差异模式表明，我们对eNOS基因的发现可能广泛相关。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节的贡献。我们还将确定特别富含EC的大型互助非编码RNA。我们预计我们的研究将为您提供新的见解表观遗传途径对EC特异性基因表达全球模式的贡献