Epigenetic Transgenerational Endocrine Disruptor Actions

描述（由申请人提供）：环境因素的跨代效应，如杀虫剂，塑料和杀真菌剂，显着放大了这些化合物的影响和健康危害。这些化合物的跨代作用需要种系的可遗传表观遗传改变。这种跨代表观遗传现象预计是成人发病疾病病因学的一个重要方面，并表明祖先的环境暴露可能会影响疾病流行病学。目前的建议旨在研究这种跨代现象和潜在的表观遗传机制。使用的模型内分泌干扰物将是杀真菌剂vinclozolin，一种抗雄激素化合物，在远交啮齿动物（即大鼠）系统中进行了研究。以前，我们证明，乙烯氯唑啉暴露在胚胎性腺性别决定促进男性生殖细胞的表观遗传重编程，然后诱导跨代成年男性不育症，前列腺疾病，肾脏疾病，免疫异常和肿瘤发展的疾病状态。本提案的目的是进一步深入了解这种内分泌干扰物在诱导这种跨代表观遗传现象方面的分子、细胞和生理（即系统生物学）作用。待检验的总体假设是，子宫内短暂暴露于内分泌干扰物乙烯氯唑啉促进雄性生殖系表观基因组的永久性重编程（即DNA甲基化），然后通过关键表观遗传调控机制的改变，转代促进成年发病疾病（例如男性不育症）。先前的研究表明，通过DNA甲基化的改变，对男性生殖系（精子）产生跨代表观遗传效应。生殖系中的这种表观遗传改变被认为随后促进了对成人许多器官系统的表观基因组和转录组的跨代效应。目前的建议旨在进一步研究这些跨代的表观遗传对男性生殖系的影响，以确定与特定成人发病疾病状态的诱导的功能关系，并揭示负责这种现象的潜在表观遗传机制。验证上述假设的实验方法包括以下具体目的：1）研究乙烯唑啉对精子表观基因组（DNA甲基化）的跨代作用，以鉴定全基因组表观遗传生物标志物。2)描述乙烯氯唑啉暴露对胎儿雄性生殖细胞表观基因组和转录组的直接和跨代影响。3)将精子表观遗传生物标志物与跨代成人发病疾病表型相关联，并研究生物标志物在与特定疾病状态相关的成人体组织转录组失调中的潜在作用。完成拟议的研究将确定环境暴露和化合物如何以跨代方式促进成人发病。将鉴定与雄性生殖系的表观遗传重编程相关的表观遗传生物标志物。这些生物标志物可能在新创建的表观遗传控制区（ECR）中促进组织转录组的跨代失调的潜在作用将被建立，并提供对成人发病疾病病因的深入了解。表观遗传生物标志物用作特定成人发病疾病状态的早期诊断标志物的潜力也将被确定。拟议的研究将更彻底地调查这种表观遗传的跨代现象，并阐明所涉及的分子机制。

DESCRIPTION (provided by applicant): Transgenerational effects of environmental factors, such as pesticides, plastics and fungicides, significantly amplify the impact and health hazards of these compounds. The transgenerational actions of these compounds require a heritable epigenetic alteration of the germline. This transgenerational epigenetic phenomenon is anticipated to be an important aspect of adult onset disease etiology, and suggests ancestral environmental exposure may influence disease epidemiology. The current proposal is designed to investigate this transgenerational phenomenon and the underlying epigenetic mechanism(s) involved. The model endocrine disruptor utilized will be the fungicide vinclozolin, an anti-androgenic compound, studied in an outbred rodent (i.e. rat) system. Previously, we demonstrated that vinclozolin exposure during embryonic gonadal sex determination promotes an epigenetic reprogramming of the male germline that then induces transgenerational adult onset disease states of male infertility, prostate disease, kidney disease, immune abnormalities and tumor development. The objective of the current proposal is to provide further insights into the molecular, cellular and physiological (i.e. systems biology) actions of this endocrine disruptor on the induction of this transgenerational epigenetic phenomenon. The overall hypothesis to be tested is that transient in utero exposure to the endocrine disruptor vinclozolin promotes a permanent reprogramming of the epigenome (i.e. DNA methylation) of the male germ line that then, through alterations in critical epigenetic regulatory mechanism(s), transgenerationally promotes adult onset disease (e.g. male infertility). Previous studies have shown a transgenerational epigenetic effect on the male germ line (sperm) through alterations in DNA methylation. This epigenetic alteration in the germ line is proposed to subsequently promote transgenerational effects on the epigenomes and transcriptomes of numerous organ systems in the adult. The current proposal is designed to further investigate these transgenerational epigenetic effects on the male germ line to determine the functional relationship with the induction of specific adult onset disease states, and to reveal the underlying epigenetic mechanisms responsible for this phenomenon. The experimental approach to test the above hypothesis consists of the following specific aims: 1) Investigate the transgenerational actions of vinclozolin on the sperm epigenome (DNA methylation) to identify genome-wide epigenetic biomarkers. 2) Characterize the direct and transgenerational effects of vinclozolin exposure on the fetal male germ cell epigenome and transcriptome. 3) Correlate the sperm epigenetic biomarkers with transgenerational adult onset disease phenotypes, and investigate the potential role of the biomarkers in the dysregulation of adult somatic tissue transcriptomes associated with specific disease states. Completion of the proposed research will determine how environmental exposures and compounds may promote adult onset disease in a transgenerational manner. The epigenetic biomarkers associated with the epigenetic reprogramming of the male germ line will be identified. The potential role these biomarkers may have in newly created epigenetic control regions (ECR) to promote transgenerational dysregulation of tissue transcriptomes will be established and provide insight into the etiology of adult onset disease. The potential for epigenetic biomarkers to be used as early stage diagnostic markers for specific adult onset disease states will also be established. The proposed research will more thoroughly investigate this epigenetic transgenerational phenomenon and elucidate the molecular mechanisms involved.