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

• 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
• 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

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 邻苯二甲酸酯，一类用于塑料和个人护理产品的内分泌干扰化合物(EDCs)， 是无处不在的环境污染物，导致人类广泛接触。流行病学数据 与父亲的邻苯二甲酸酯有关，导致生殖健康不良，包括精子质量差，最近， 随着怀孕时间的延长，−暗示了一种精子来源的效应。不断增长的令人信服的数据 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，反过来可以在受精时传递信息，影响健康和 后代的发育。众所周知，男性生殖细胞的产前表观遗传重新编程是一种 特别容易受到邻苯二甲酸盐等环境暴露的窗户。研究现已浮出水面 表明在生殖细胞发育过程中还存在其他这样的易感窗口。我们的目标是 确定成人接触空气的影响 邻苯二甲酸二乙基己酯(DEHP)、邻苯二甲酸二丁酯(DBP)和 DEHP和DBP混合物对精子DNA甲基化和精子DNA甲基化的影响 精子来源变化的持久性 胚胎和F1成体生殖细胞的DNA甲基化编程。我们将曝光成人生殖 日龄雄性C57BL/6J小鼠分别给予0、2.5 mg/kg/d的DEHP、2.5 mg/kg/d的DBP及其混合物(DEHP和 DBP，每次2.5毫克/公斤/天)，持续70天--大约两个精子发生周期。附睾精子会 从每组小鼠身上收集，并将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-- 对产生的F1杂交胚胎进行跨基因组的特定分析。我们将执行RNA-SEQ和WGBS 在原肠E6.5胚胎的胚胎和胎盘细胞中检测父本DEHP和/或 在每个不同的血统上都有DBP。最后，我们将分析精子质量参数、卵泡分期和 雄性和雌性F1后代的生殖激素。我们还将在F1上进行RNA-Seq和WGBS 精子和卵母细胞。这项提出的研究有望通过精子识别表观遗传学。 先入为主的男性EDC暴露和后代生殖健康的影响的甲基化。 理解精子表观遗传学在暴露-疾病范例中的参与将有助于 开发新的治疗和预防方法的转化性研究途径 对生殖健康不利。