Effects of Phthalates on Trophoblast Differentiation: From Biology to Biomarkers

项目概要/摘要 我寻求资金来扩展我在流行病学的两个新方向的培训-即人类胎盘生物学 和质谱分析。学习这些基于实验室的方法将使我更接近于完成我的 成为一名独立的学术研究者，并提供新的见解和方法， 低剂量长期暴露于内分泌干扰化合物及其在健康疾病中的作用的研究 这是在怀孕早期发生的。在K99阶段，在Susan Fisher博士的主要指导下，我 将接受滋养层（Tb）干细胞生物学的培训，最先进的基于微阵列的方法， 用于在全球水平上分析基因表达，包括用于数据分析的生物信息学方法。 基于我最初的博士后研究结果，我认为结核分枝杆菌，携带结核分枝杆菌的特化细胞， 胎盘的许多最重要的功能，在怀孕早期暴露于邻苯二甲酸酯， 对胎盘发育和功能的不良后果。我们将测试关于临床的假设 结果和生物学参数，包括人类的分子，细胞和形态学测量 胎盘的发育和功能。具体地说，我将进行实验来验证培养的假设， 人结核病干细胞将在基因表达模式中显示剂量依赖性变化， 暴露于环境相关剂量的邻苯二甲酸二（2-乙基己基）酯和邻苯二甲酸二正丁酯（目标1）。 在R 00阶段，我将在一项对孕妇的纵向研究中验证这一理论，即胎盘基因 作为邻苯二甲酸酯体外暴露的结果而差异表达的蛋白质， 剂量依赖性的方式，在体内（目的2）。作为这一目标的一部分，我将评估 产前暴露的常规剂量计、尿邻苯二甲酸酯代谢物浓度和 目标是胎盘组织我的主要导师和妇产科，质谱学， 生物统计学和流行病学将指导我学习所需方法的过程， 实现本提案提出的目标。在这个培训项目结束时， 我和其他研究人员可以使用新的方法和工具来询问有关 妊娠期间邻苯二甲酸酯暴露对胎盘功能的影响， 因此，人类发展。这项工作很重要，因为邻苯二甲酸酯，可以破坏细胞， 组织分化，是公认的生殖和发育毒物在啮齿动物;然而， 这些发现与人类的相关性还没有得到很好的理解。与邻苯二甲酸酯诱导的特异性生物标志物 胎盘损伤，它将成为可能，以确定高风险怀孕，并提供机会， 在人口一级进行预防，并可能在保健提供系统一级进行干预， 改善胎盘和胎儿结局。

Project Summary/Abstract I seek funding to extend my training in epidemiology in two new directions-namely, human placental biology and mass spectrometry. Learning these laboratory-based methods will move me closer to accomplishing my long-term goals of becoming an independent academic investigator and offering new insights and methods to research on low dose chronic exposures to endocrine disrupting compounds and their role in health disorders that originate in early pregnancy. During the K99 phase, under the primary mentorship of Dr. Susan Fisher, I will receive training in trophoblast (Tb) stem cell biology, state-of-the-art microarray-based methodologies that are used to analyze gene expression at a global level, including bioinformatics approaches for data analysis. Building on the results of my initial post-doctoral research, I theorize that Tbs, the specialized cells that carry out many of the placenta's most important functions, are exposed to phthalates early in pregnancy with adverse consequences on placental development and function. We will test hypotheses regarding clinical outcomes and biological parameters that include molecular, cellular, and morphologic measures of human placental development and function. Specifically, I will conduct experiments to test the hypothesis that cultured human Tb stem cells will show dose-dependent changes in patterns of gene expression when they are exposed to environmentally relevant doses of di-(2-ethylhexyl) phthalate and di-n-butyl phthalate (Aim 1). During the R00 phase, I will test the theory, in a longitudinal study of pregnant women, that placental genes that are differentially expressed as a consequence of phthalate exposure in vitro are similarly regulated, in a dose-dependant manner, in vivo (Aim 2). As part of this aim, I will evaluate the correlation between the conventional dosimeter of prenatal exposure, urinary phthalate metabolite concentrations, and the dose to the target placental tissue. My primary mentor and co-mentors in obstetrics/gynecology, mass spectrometry, biostatistics, and epidemiology will guide the process whereby I learn the methods that are required to accomplish the goals set forth in this proposal. At the conclusion of this training program I will have developed new methodologies and tools that I and other investigators can use to ask more directed questions about the consequences of phthalate exposures during pregnancy in terms of alterations in placental function and consequently, human development. This work is important because phthalates, which can disrupt cell and tissue differentiation, are well established as reproductive and developmental toxicants in rodents; yet the relevance of these finding to humans is not well understood. With biomarkers specific to phthalate-induced placental damage, it will become possible to identify high-risk pregnancies and provide opportunities for prevention, at the population level, and possibly intervention at the level of health care delivery systems to improve placental and fetal outcomes.