Effects of PAHs on the Epigenome and Differentiation Capacity of Embryonic and Ne

PAHs对胚胎和神经细胞表观基因组和分化能力的影响

• 批准号: 8387978
• 负责人: Raymond David Hawkins
• 金额: $ 23.18万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8387978
• 关键词: 7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide; Affect; Antibodies; Aromatic Polycyclic Hydrocarbons; Biological Markers; Biological Models; Cells; ChIP-seq; Cytosine; DNA; DNA Adducts; DNA Methylation; Defect; Development; Disease; Embryo; Environment; Epigenetic Process; Event; Exposure to; Genome; Genomics; Germ Layers; Goals; Human Development; Lead; Location; Maps; Methods; Methylation; Modification; Monitor; Mutation; Neurologic; Pattern; Population; Protocols documentation; Pyrenes; Research Personnel; Role; Site; Stem cells; System; Time; Toxic Environmental Substances; adduct; benzo(a)pyrene-7,8-dihydrodiol-9,10-epoxide-DNA; cell fate specification; cell type; epigenomics; genome-wide; histone modification; human embryonic stem cell; nerve stem cell; novel; progenitor; response

DESCRIPTION (provided by applicant): Human embryonic stem cells (hESCs) provide an excellent model system for studying cellular differentiation and human development. These cells can give rise to cell types from all three germ layers, with numerous protocols for cell-specific populations. However, this system has been under-utilized to determine the effects of the environment, specifically regarding exposure to toxic agents that can have a detrimental effect. Here hESCs will be used to determine how polycyclic aromatic hydrocarbons (PAHs) alter the epigenome of hESCs and ESC-derived neural progenitor cells (NPCs) and if this in turn affects their differentiation capacity. As proof of principle the focus will be on the widely studied 7,8 dihydroxy-9,10-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene or BPDE. BPDE is known to form DNA adducts that can lead to mutational changes, but additional evidence has shown that such adducts are often targeted to methylated CpGs. As has been previously demonstrated, hESCs have very unique epigenomes through the global mapping and analysis of histone modifications and DNA methylation. It was determined that cytosine methylation patterns change in response to different concentrations of BPDE. Therefore, the intent here is to examine if exposure has a detrimental effect on the ability of hESCs and NPCs to differentiate, with regards to timing or efficiency of lineage specification. In addition, it is important to determine the specific genomic sites of BPDE-DNA adducts. Expertise in ChIP-seq will be leveraged to develop a global identification method of these adducts using antibodies against BPDE-DNA. PUBLIC HEALTH RELEVANCE: The investigators will establish human embryonic stem cells and neural progenitor cells as a system for determining the environmental epigenomic effects on these progenitor populations by exposure to the polycyclic aromatic hydrocarbon BPDE. They will determine if this affects the differentiation capacity and results in changes to global DNA methylation. The investigators will also develop a method to determine the genome-wide location of DNA adducts caused by BPDE, called BPDE-DNA-seq.

描述（由申请人提供）：人类胚胎干细胞（hESC）为研究细胞分化和人类发育提供了极好的模型系统。这些细胞可以产生来自所有三个胚层的细胞类型，并针对细胞特异性群体提供多种方案。然而，该系统在确定环境影响方面尚未得到充分利用，特别是在接触可能产生有害影响的有毒物质方面。在此，hESC 将用于确定多环芳烃 (PAH) 如何改变 hESC 和 ESC 衍生的神经祖细胞 (NPC) 的表观基因组，以及这是否反过来影响它们的分化能力。作为原理证明，重点将放在广泛研究的 7,8 二羟基-9,10-环氧-7,8,9,10-四氢苯并[a]芘或 BPDE 上。已知 BPDE 会形成可导致突变变化的 DNA 加合物，但更多证据表明，此类加合物通常针对甲基化 CpG。正如之前所证明的，通过组蛋白修饰和 DNA 甲基化的全局作图和分析，hESC 具有非常独特的表观基因组。已确定胞嘧啶甲基化模式会响应不同浓度的 BPDE 而变化。因此，本文的目的是检查暴露是否对 hESC 和 NPC 的分化能力（在谱系规范的时间或效率方面）产生不利影响。此外，确定特定的基因组也很重要。 BPDE-DNA 加合物位点。将利用 ChIP-seq 方面的专业知识，使用 BPDE-DNA 抗体开发这些加合物的全球鉴定方法。 公共健康相关性：研究人员将建立人类胚胎干细胞和神经祖细胞作为一个系统，通过暴露于多环芳烃 BPDE 来确定环境表观基因组对这些祖细胞群的影响。他们将确定这是否会影响分化能力并导致整体 DNA 甲基化的变化。研究人员还将开发一种方法来确定 BPDE 引起的 DNA 加合物的全基因组位置，称为 BPDE-DNA-seq。