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：
9549064
负责人：
J. Richard Pilsner
金额：
$ 45.95万
依托单位：
UNIVERSITY OF MASSACHUSETTS AMHERST
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-01 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9549064
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 Pregnancy Prevention Regulation Reporting Reproductive Health Research 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 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.

项目概要邻苯二甲酸盐是一类用于塑料和个人护理产品的内分泌干扰化合物 (EDC)，是普遍存在的环境污染物，导致人类广泛接触。表明父系邻苯二甲酸盐对生殖健康不利，包括精子质量差，最近，怀孕时间更长——后者表明精子产生的效应越来越多，令人信服。表明环境暴露可以通过以下方式体现在发育中的雄性生殖细胞中表观遗传标记反过来又可以在受精时传递信息，从而影响健康和发展轨迹众所周知，雄性生殖细胞的产前表观遗传重编程是一个过程。现在已经出现了特别容易受到邻苯二甲酸盐等环境暴露影响的窗口。证明生殖细胞发育过程中存在其他此类易感窗口。确定成人接触的影响邻苯二甲酸二(2-乙基己基)酯 (DEHP)、邻苯二甲酸二正丁酯 (DBP) 和 DEHP和DBP混合物对精子DNA甲基化的影响精子衍生的变化的持续性孕体和 F1 成体生殖细胞的 DNA 甲基化编程我们将揭示成体生殖细胞。年龄雄性 C57BL/6J 小鼠，给予 0、2.5 mg/kg/天 DEHP、2.5 mg/kg/天 DBP 及其混合物（DEHP 和 DBP（每次 2.5 毫克/公斤/天）持续 70 天——大约两个附睾精子发生周期。从每组小鼠中收集并进行全基因组亚硫酸氢盐测序（WGBS）分析。接下来，暴露的 C57BL/6J 雄性将与未暴露的 DBA/2J 雌性交配，这将允许等位基因- 我们将对产生的 F1 杂交胚胎的基因组进行特异性分析。在原肠胚 E6.5 胚胎的胚胎和胎盘细胞中测定父本 DEHP 和/或的影响最后，我们将分析每个不同谱系的精子质量参数、卵母细胞分期和我们还将对 F1 后代进行 RNA 测序和 WGBS。这项研究预计将通过精子来确定表观遗传学。甲基化对孕前男性 EDC 暴露和后代生殖健康的影响。了解精子表观遗传学在暴露疾病范式中的参与将为我们提供信息开发治疗和预防新方法的转化研究途径不利生殖健康。