Male preconception phthalates and offspring embryo and sperm allele-specific methylome programming.

男性孕前邻苯二甲酸盐和后代胚胎和精子等位基因特异性甲基化组编程。

基本信息

批准号：
10414482
负责人：
J. Richard Pilsner
金额：
$ 43.31万
依托单位：
WAYNE STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-01 至 2024-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10414482
关键词：
Adult Adult Children Affect Alleles Animal Model Bioinformatics Biological Models Cells Conceptus DNA Methylation Data Development Dibutyl Phthalate Diethylhexyl Phthalate Disease Embryo Embryonic Development Endocrine Disruptors Environment Environmental Exposure Environmental Health Environmental Pollution Epigenetic Process Exposure to Female Fertilization Gene Expression Genetic Genome Genomics Germ Cells Goals Health Heritability Human Hybrids Interdisciplinary Study Knowledge Link Massachusetts Methylation Mus Nutritional Oocytes Partner in relationship Placenta Plastics Prevention Regulation Reporting Reproductive Health Research Spermatocytes Spermatogenesis Staging Time Tissues Toxicology Translational Research Universities androgenic animal data behavioral health bisulfite sequencing embryo tissue epidemiologic data exposed human population fetal frontier male methylome multidisciplinary novel strategies offspring personal care products phthalates prenatal reproductive reproductive hormone research and development sperm cell sperm quality time-to-pregnancy toxicant transcriptome sequencing whole genome

项目摘要

Project Summary Phthalates, a class of endocrine disrupting compounds (EDCs) used in plastics and personal care products, are ubiquitous environmental contaminants resulting in widespread human exposure. Epidemiologic data implicate paternal phthalates with adverse reproductive health including poor sperm quality, and more recently, with longer time to pregnancy − the latter suggests a sperm-derived effect. A growing body of compelling data demonstrates that environmental exposures can be embodied within the developing male germ cell through epigenetic marks, which in turn, can impart information at fertilization to affect the trajectory of health and development of offspring. It is known that prenatal epigenetic reprogramming of male germ cells is a particularly susceptible window to environmental exposures such as phthalates. Research has now emerged demonstrating that other such susceptible windows exist during germ cell development. Our objective is to determine the effects of adult exposure to di(2-ethylhexyl) phthalate (DEHP), di-n-butyl phthalate (DBP) and the mixture of DEHP and DBP on sperm DNA methylation and the persistence of sperm-derived changes on DNA methylation programming of the conceptus and F1 adult germ cells. We will exposure adult reproductive age male C57BL/6J mice with 0, 2.5 mg/kg/day of DEHP, 2.5 mg/kg/day of DBP, and its mixture (DEHP and DBP each at 2.5 mg/kg/day) for 70 days – approximately two spermatogenesis cycles. Epididymal sperm will be collected from mice in each group and will be analyzed for whole genome bisulfite sequencing (WGBS). Next, exposed C57BL/6J males will be mated with unexposed DBA/2Jfemales, which will allow for allele- specific analysis across the genome in the F1 hybrid embryos produced. We will perform RNA-seq and WGBS in embryonic and placental cells of gastrulating E6.5 embryos to determine the effects of paternal DEHP and/or DBP on each distinct lineage. Finally, we will analyze sperm quality parameters, oocyte follicle staging and reproductive hormones in male and female F1 offspring. We will also perform RNA-seq and WGBS on F1 sperm and oocytes. This research proposed is expected to identify an epigenetic understanding via sperm methylation of the effects of preconception male EDC exposure and offspring reproductive health. Understanding the involvement of sperm epigenetics within the exposure-disease paradigm will inform avenues of translational research for the development of novel approaches for the treatment and prevention of adverse reproductive health.

项目摘要邻苯二甲酸盐，一类用于塑料和个人护理产品的内分泌干扰化合物（EDC），无处不在的环境污染物会导致人类暴露宽度。流行病学数据暗示具有不良繁殖健康（包括精子质量差）的父亲邻苯二甲酸盐，最近随着怀孕的时间，后者表明精子衍生的作用。越来越多的引人入胜的数据证明环境暴露可以通过表观遗传标记又可以在受精时传达信息，以影响健康的轨迹后代的发展。众所周知，雄性生殖细胞的产前表观遗传重编程是一个特别容易受到环境暴露（例如邻苯二甲酸盐）的窗口。研究现已出现证明在生殖细胞发育过程中存在其他这种易感窗户。我们的目标是确定成人接触的影响 DI（2-乙基己基）邻苯二甲酸酯（DEHP），二丁基苯甲酸酯（DBP）和 DEHP和DBP在精子DNA甲基化和DBP的混合物以及精子衍生的变化的持久性概念和F1成人生殖细胞的DNA甲基化编程。我们将接触成人生殖年龄雄性C57BL/6J小鼠，DEHP的0、2.5 mg/kg/天，DBP的2.5 mg/kg/day及其混合物（DEHP和DEHP DBP分别为2.5 mg/kg/天）70天 - 大约两个精子发生周期。附子精子会从每组中的小鼠中收集，并将分析全基因组Bisulfite测序（WGB）。接下来，暴露的C57BL/6J男性将与意外的DBA/2Jfemales成为伴侣，这将允许使用 - 在产生的F1杂种胚胎中跨基因组的特定分析。我们将执行RNA-Seq和WGB 在胃组织E6.5胚胎的胚胎和斑点细胞中，以确定父亲DEHP和/或每个不同的谱系上的DBP。最后，我们将分析精子质量参数，卵母细胞分期和男性和女性F1后代的生殖恐怖。我们还将在F1上执行RNA-Seq和WGB 精子和卵母细胞。这项提出的研究预计将通过精子确定表观遗传学的理解雄性EDC暴露和后代生殖健康的影响的甲基化。了解精子表观遗传学在暴露范围范式中的参与将告知转化研究的途径，用于开发新方法的治疗和预防方法不良复制健康。