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出色的模型系统。这些细胞可以从所有三个细菌层中产生细胞类型，并具有许多用于细胞特异性种群的方案。但是，该系统已被详尽地确定环境的影响，特别是关于暴露于可能产生不利影响的有毒药物的影响。这里将使用hESC来确定多环芳烃（PAHS）如何改变hESC和ESC衍生的神经祖细胞（NPC）的表观基因组（NPC），并且如果这反过来会影响其分化能力。作为原则的证明，焦点将放在经过广泛研究的7,8二羟基-9,10-Epoxy-7,8,9,9,10-四氢苯甲酸[a] pyrene或bpde。已知BPDE形成可能导致突变变化的DNA加合物，但其他证据表明，这种加合物通常针对甲基化的CPG。如前所述，通过对组蛋白修饰和DNA甲基化的全局映射和分析，hESC具有非常独特的表观基因概。据确定，胞嘧啶甲基化模式会因不同浓度的BPDE而发生变化。因此，这里的目的是检查暴露是否对hESC和NPC分化的能力有害影响，涉及谱系规范的时间或效率。另外，确定特定基因组很重要 BPDE-DNA加合物的位点。将利用CHIP-SEQ方面的专业知识来使用针对BPDE-DNA的抗体开发这些加合物的全球识别方法。 公共卫生相关性：研究人员将通过暴露于多环芳族烃BPDE来确定对这些祖细胞种群的环境表观基因组影响的系统。他们将确定这是否影响分化能力并导致全局DNA甲基化的变化。研究人员还将开发一种确定由BPDE引起的DNA加合物的全基因组位置的方法，称为BPDE-DNA-SEQ。