Phthalate Exposure, Sperm DNA Methylation and Early-life Development

总结 这个K22职业发展奖的目的是促进J.理查德博士的职业发展 皮尔森是一位跨学科的分子流行病学家。一个长期的研究目标是描述 表观遗传学作为连接母亲和父亲环境的机制途径的中间步骤 生殖成功和随后的后代健康和发育的风险。作为一个新任命的 助理教授，我已经提供了必要的设施和资源，以建立一个强大的高- 该实验室是一个高通量实验室，可进行专为大型人群研究量身定制的表观遗传分析。的目的 K22的应用是建立一项队列研究，对250对寻求体外受精的低生育力夫妇进行检查， 父亲邻苯二甲酸酯暴露、精子表观遗传学和生殖健康之间的关联。本 最后，我已经确定了一些具体的领域，在这些领域，额外的发展将提高我作为一个独立的人的成功 研究员这些包括：1）内分泌干扰化合物（EDCs）; 2）精子生物学和男性生殖 健康; 3）队列和生物储存库的实施和4）表观遗传信息学和计算生物学。 邻苯二甲酸酯是一种EDC，是普遍存在的环境污染物，用于塑料和个人护理 产品.邻苯二甲酸酯暴露与多种生殖结果相关，包括男性 生殖健康、生育结果和后代神经发育;然而， 这些关联尚未明确阐明。表观遗传重编程对精子至关重要 成熟，并代表了对环境诱导的表观遗传错误的敏感性的关键窗口， 反过来又可能影响生殖健康。新的动物和人类数据表明， 健康与精子的表观遗传失调有关。此外，最近的证据表明， 保留在成熟精子中的表观遗传信息在发育和印记基因中富集， 精子的表观遗传模式可能对受精后的胚胎发育很重要。实现 我们的研究目标是：1）检查父亲尿邻苯二甲酸酯暴露与 精子DNA甲基化在印迹基因和Illumina的450 K表观基因组阵列; 2）检查影响 父亲邻苯二甲酸酯暴露对精子质量和胚胎发育的影响; 3）使用统计中介， 检查精子甲基化是否是父亲邻苯二甲酸酯影响精子和胚胎质量的途径。 这项研究是创新的，因为：1）虽然许多人类的表观遗传学研究受到DNA的阻碍， 混合细胞类型（例如，白细胞），我们的研究利用单细胞类型-精子生殖细胞; 2）它将提供， 据我们所知，第一个关于邻苯二甲酸酯暴露与精子DNA之间关系的人类数据 通过使用靶向基因和表观基因组方法来抑制甲基化;以及3）它可以挑战 通过提供关于父亲对生殖健康的环境贡献的新数据， 繁殖成功。

Summary The purpose of this K22 Career Development Award is to foster the career development of Dr. J. Richard Pilsner, an interdisciplinary molecular epidemiologist. A long-term research goal is to delineate the role of epigenetics as an intermediate step in the mechanistic pathway linking maternal and paternal environmental exposures to reproductive success and subsequent offspring health and development. As a newly appointed Assistant Professor, I have been provided the necessary facilities and resources to establish a robust high- throughput laboratory for epigenetic analyses tailored for large population-based studies. The objective of this K22 application is to establish a cohort study of 250 subfertile couples seeking in vitro fertilization to examine the association between paternal phthalate exposure, sperm epigenetics, and reproductive health. To this end, I have identified specific areas where additional development will enhance my success as an independent researcher. These include: 1) endocrine disruptor compounds (EDCs); 2) sperm biology and male reproductive health; 3) cohort and biorepository implementation and 4) epigenetic informatics and computational biology. Phthalates, an EDC, are ubiquitous environmental contaminants that are used in plastics and personal care products. Phthalate exposure is associated with a diverse set of reproductive outcomes, including male reproductive health, birth outcomes, and offspring neurodevelopment; however, a mechanistic understanding of these associations has not been clearly elucidated. Epigenetic reprogramming is essential for sperm maturation and represents a critical window of susceptibility to environmentally-induced epigenetic errors that may, in turn, influence reproductive health. Emerging animal and human data indicate that male reproductive health is associated with epigenetic dysregulation in sperm. Moreover, recent evidence indicates that epigenetic information retained in mature sperm is enriched at developmental and imprinted genes, signifying that epigenetic patterns in sperm may be important for embryonic development after fertilization. To achieve our research objectives, we will: 1) examine the relationships between paternal urinary phthalate exposure on sperm DNA methylation at imprinted genes and by Illumina's 450K epigenomic array; 2) examine the influence of paternal phthalate exposure on sperm quality and embryo development; 3) use statistical mediation to examine if sperm methylation is a pathway by which paternal phthalates influence sperm and embryo quality. This study is innovative because: 1) while many epigenetic studies in humans are hampered by DNA from mixed cell types (e.g., leukocytes), our study utilizes a single cell type - sperm germ cells; 2) it will provide, to our knowledge, the first human data on the association between phthalate exposure and sperm DNA methylation through the use of both targeted gene and epigenomic approaches; and 3) it may challenge the current dogma of reproductive health by providing novel data on paternal environmental contributions to reproductive success.