Chromatin modifications and DNA methylation during early lung development

早期肺部发育过程中的染色质修饰和 DNA 甲基化

基本信息

批准号：
7391424
负责人：
Maria Isabel Ramirez
金额：
$ 43.91万
依托单位：
BOSTON UNIVERSITY MEDICAL CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-12-01 至 2012-11-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7391424
关键词：
Antibodies Binding Biological Assay Breeding Cell Differentiation process Cell Lineage Cells Chromatin Chromatin Remodeling Factor Commit Complex DNA DNA Binding DNA Methylation DNA Modification Methylases DNA amplification Deoxycytidine Development Dominant-Negative Mutation EZH2 gene Embryo Endoderm Epigenetic Process Epithelium Evaluation FGFR2 gene Gene Activation Gene Expression Gene Expression Regulation Gene Targeting Genes Genetic Transcription Genome Genomics Goals Histones In Vitro Knowledge Lentivirus Vector Link Liver Lung Mesenchyme Messenger RNA Methylation Microarray Analysis Modification Mus Organ Organogenesis Pattern Play Polycomb Post-Translational Protein Processing Primitive foregut structure Primordium Proteins RNA Polymerase II Regulation Role System Testing Tissues Undifferentiated base bisulfite brahma brahma protein chromatin immunoprecipitation chromatin remodeling day embryonic stem cell enzyme activity genetic regulatory protein in vivo lung development programs promoter protein expression recombinase small hairpin RNA stem

项目摘要

The goal of this project is to investigate the role of chromatin modifications and DMA methylation in activation or silencing of gene expression during initiation of lung development. Chromatin modifications and DNA methylation in conjunction with DNA binding factors regulate transcription to achieve cell specific gene expression. These mechanisms play a defined role in developmental programs, and have been shown to be involved in the establishment of the specific cell lineages that will give rise to various organs, including foregut derivatives such as the liver. Little is known about how these mechanisms influence gene expression in the developing lung. In a microarray analysis of the developing foregut carried during the previous period of this Program Project, we identified significant changes in the expression of genes associated with chromatin remodeling and DNA methylation, coincident with the formation of the lung primordium. The known important role of chromatin remodeling in cell fate decisions and differentiation led us to hypothesize that chromatin remodeling and DNA methylation play a regulatory role in initiation of lung development by establishing spatial and temporal patterns of gene expression in the embryonic foregut. Thus we propose to: 1) identify changes in epigenetic modifications of selected lung gene promoters during initiation of lung development in vivo and in cultured ES cells, 2) search for targets of activating chromatin remodeling complexes in embryonic lung cells, 3) analyze the function of chromatin remodeling genes and DNA methylation on the transcriptional activity of lung promoters in cells, and in wild type and genetically modified lungs. The proposed studies include the characterization of chromatin modifications of lung gene promoters by chromatin immunoprecipitation, DNA methylation and chromatin accessibility analyses in vitro and in vivo, a global analysis of new targets of epigenetic gene activation, and the functional evaluation of chromatin modification genes in lung gene development using foregut cultures and genetically modified mice. Our findings will increase the knowledge of how the early patterns of lung expression are regulated in vivo and in ES cells.

该项目的目的是研究染色质修饰和DMA甲基化在激活中的作用或在肺发育开始过程中基因表达的沉默。染色质修饰和DNA 与DNA结合因子结合的甲基化调节转录以实现细胞特异性基因表达。这些机制在发展计划中起着明确的作用，已被证明是参与建立特定细胞谱系，这些谱系将引起各种器官，包括肝之类的前肢衍生物。这些机制如何影响基因知识知之甚少发育中的肺表达。在对在此期间进行的发育前表面的微阵列分析该程序项目的上一个时期，我们确定了基因表达的重大变化与染色质重塑和DNA甲基化相关，与肺的形成一致原始。染色质重塑在细胞命运决策和分化LED中的已知重要作用我们假设染色质重塑和DNA甲基化在肺的启动中起着调节作用通过在胚胎前肠中建立基因表达的空间和时间模式来发展。因此，我们建议：1）确定选定肺基因启动子的表观遗传修饰的变化在体内和培养的ES细胞中肺发育的启动，2）搜索激活染色质的靶标胚胎肺细胞中的重塑复合物，3）分析染色质重塑基因的功能和 DNA甲基化对细胞中肺启动子的转录活性以及野生型和遗传的转录活性修饰的肺。拟议的研究包括肺基因染色质修饰的表征通过染色质免疫沉淀，DNA甲基化和染色质可及性分析的启动子在体外进行在体内，对表观遗传基因激活的新靶标的全球分析以及对功能评估使用前述培养物和基因修饰的肺基因发育中的染色质修饰基因老鼠。我们的发现将增加有关如何调节肺表达的早期模式的知识体内和ES细胞中。