Mechanisms of transgenerational epigenetic inheritance

跨代表观遗传机制

• 批准号: 9899105
• 负责人: Victor G. Corces
• 金额: $ 34.73万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-04-01 至 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9899105
• 关键词: 3-Dimensional; ATAC-seq; Adipocytes; Adult; Affect; Antibodies; Architecture; Binding; Binding Sites; Cell Differentiation process; Cell Nucleus; ChIP-seq; Chromatin; Chromosomes; Complex; DNA; DNA Methylation; DNA-Binding Proteins; Data; Development; Enhancers; Environment; Environmental Risk Factor; Epigenetic Process; Exposure to; Female; Fertilization; Frequencies; Gene Expression; Generations; Genes; Genetic Transcription; Genome; Germ Lines; Histones; Human; Hypersensitivity; Individual; Link; Location; Maintenance; Mammals; Maps; Mass Spectrum Analysis; Mediating; Mediator of activation protein; Metabolic Diseases; Modification; Mus; Nature; Nucleosomes; Obese Mice; Obesity; Onset of illness; Outcome; Paint; Parents; Pattern; Phenotype; Positioning Attribute; Pregnancy; Procedures; Protamines; Proteins; Protocols documentation; RNA Polymerase II; Resolution; Site; Somatic Cell; Spermatids; Spermatogenesis; Spermiogenesis; Structure of primordial sex cell; Suggestion; Techniques; Tissues; Transcription Alteration; Transcription Initiation Site; Untranslated RNA; Work; base; bisphenol A; bisulfite sequencing; chromosome conformation capture; cohesin; condensin; embryo cell; epigenome; experimental study; histone modification; insight; male; mouse model; next generation; novel; offspring; pluripotency; pregnant; sperm cell; stem cells; transcription factor; transcriptome sequencing; transgenerational epigenetic inheritance; transmission process

Project Summary/Abstract Environmental factors increase the frequency of metabolic disorders. The mechanisms by which these epiphenotypes are transmitted between the exposed and subsequent generations through the paternal germline remains poorly understood. The nuclei of mammalian sperm are highly condensed, the DNA is mostly covered by protamines with only a few retained nucleosomes, and epigenetic information stored in the form of DNA methylation is quickly erased from paternal chromosomes shortly after fertilization. Experiments carried out in our lab suggest a more complex picture of mouse sperm, suggesting the presence of multiple histone modifications, nucleosomes positioned around transcription start sites and transcription factor binding sites, presence of CTCF, cohesin, condensin, FoxA1, Oct4, Nanog, Mediator and RNAPII phosphorylated in Ser2. We also find thousands of enhancers and super-enhancers in a poised or active epigenetic state based on the presence of both specific histone modifications and transcription factors. This information suggests that mammalian sperm contain a rich and complex palette of epigenetic information that could be altered by environmental factors to paint novel phenotypic outcomes in the next generation. In this application, we propose to carry out experiments to dissect the mechanisms by which epigenetic information is established and altered by the environment during male germline development, how this information is stored in sperm, and how it is transmitted to the somatic cells of adult tissues of the next generation. To accomplish this we will use obese mice who are 4th generation descendants of females exposed to Bisphenol A (BPA) during pregnancy. We will use mass spectrometry to identify a wide range of transcription factors present in sperm, and ChIP-seq to examine differences in the distribution of these transcription factors in the sperm of control versus obese mice. We will then use Chromosome Conformation Capture techniques to examine the consequence of alterations in transcription factor binding on the 3D architecture of mouse sperm. To understand how and when differences in the epigenome of control and obese mice are established, we will examine the effect of BPA treatment on transcription and transcription factor distribution using RNA-seq, ATAC-seq and ChIP-seq in primordial germ cells and spermatogonial stem cells. Similar analyses will be performed in adipocytes of obese mice in order to understand which of these epigenetic alterations are maintained in adult tissues and may be responsible for the observed obese phenotype. Results from this work will give critical insights into the mechanisms by which alterations in the epigenome are established and transmitted between generations.

项目总结/摘要 环境因素会增加代谢紊乱的发生率。的机制 这些表型通过以下途径在暴露者和后代之间传递： 父系生殖系仍然知之甚少。哺乳动物精子的细胞核 浓缩后，DNA大部分被鱼精蛋白覆盖，只有少数保留的核小体， 而以DNA甲基化形式储存的表观遗传信息很快就从父亲的 受精后不久的染色体。我们实验室的实验表明， 小鼠精子的复杂图片，表明存在多种组蛋白修饰， 位于转录起始位点和转录因子结合位点周围的核小体， 存在CTCF、粘附素、缩合素、FoxA 1、Oct 4、Nanog、介体和RNAPII 丝氨酸2磷酸化。我们还发现了成千上万的增强子和超级增强子 或基于存在特异性组蛋白修饰和 转录因子这一信息表明，哺乳动物的精子含有丰富的， 复杂的表观遗传信息调色板，可以被环境因素改变， 下一代的新表型结果。在本申请中，我们建议执行 实验，以剖析机制，其中表观遗传信息是建立和 在男性生殖系发育过程中，这些信息是如何被储存在 精子，以及它是如何传递给下一代成年组织的体细胞的。到 为了实现这一点，我们将使用肥胖小鼠，这些小鼠是暴露于 双酚A（BPA）在怀孕期间我们会用质谱仪来鉴定 精子中存在的转录因子，以及ChIP-seq来检查分布的差异 这些转录因子在对照组和肥胖组小鼠精子中的表达。然后我们将使用 染色体构象捕获技术，以检查 转录因子结合小鼠精子的3D结构。了解如何和 当对照小鼠和肥胖小鼠的表观基因组差异确定后，我们将研究 使用RNA-seq的BPA处理对转录和转录因子分布的影响， 原始生殖细胞和精原干细胞中的ATAC-seq和ChIP-seq。类似 将在肥胖小鼠的脂肪细胞中进行分析，以了解这些脂肪细胞中的哪一个 表观遗传改变维持在成体组织中，并可能是观察到的 肥胖表型这项工作的结果将提供关键的见解的机制， 表观基因组中的改变在世代之间建立和传递。