Polybrominated Diphenyl Ether Effects on Human Neuronal Development

多溴二苯醚对人类神经元发育的影响

基本信息

批准号：
8678771
负责人：
Joshua Frederick Robinson
金额：
$ 10.02万
依托单位：
UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-06-01 至 2016-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8678771
关键词：
Adult Area Behavior Bioinformatics Biological Markers Biological Models Biological Process Body Burden Cell Adhesion Molecules Cell model Cells Chemicals Childhood Critical Pathways Defect Development Disease Dose Ectoderm Embryonic Development Environmental Exposure Environmental Health Epidemiology Epigenetic Process Event Exposure to Flame Retardants Funding Future Gene Expression Gene Expression Profile Gene Targeting Generations Genes Genomics Germ Layers Goals Growth Factor Human Human Development In Vitro Investigation Knowledge Laboratories Lead Learning Measures Mentors Methods Modeling Modification Molecular Molecular Biology Molecular Profiling Mus Nervous system structure Neurodevelopmental Disorder Neuronal Differentiation Neurons Neurotoxins Outcome Oxidative Stress Pathway interactions Pattern Phase Placenta Play Population Pregnancy Process RNA Sequences Rattus Research Research Personnel Risk Rodent Model Role Seminal Series Signal Transduction Staging Statistical Methods System Techniques Testing Thyroid Hormones Toxic Environmental Substances Toxic effect Training Umbilical Cord Blood United States National Institutes of Health Work base career consumer product design developmental toxicology early life exposure embryonic stem cell environmental chemical exposure gene environment interaction human embryonic stem cell human embryonic stem cell line in vivo Model interest lentiviral-mediated nerve stem cell nervous system development neurobehavior neurobehavioral neurodevelopment neurogenesis neuron development neurotoxicity phenyl ether prenatal prenatal environmental exposure public health relevance research study screening self-renewal skills stem cell biology stem cell differentiation toxicant transcription factor

项目摘要

DESCRIPTION (provided by candidate): I seek funding to extend my training in environmental health science and developmental toxicology by acquiring additional expertise in three areas: human embryonic stem cell (hESC) biology, molecular biology, and bioinformatics. Learning laboratory-based methods and statistical approaches used in these three areas will provide me with the skills that are required to transition into an independent academic career, investigating how environmental exposures alter early human embryogenesis, and their potential contributions to childhood and adult diseases. For the K99 phase, my primary mentor is Dr. Susan Fisher, whose expertise includes using hESC models for studying human development. I will also receive valuable input from my other mentors who are leaders in the fields of human neuronal development, prenatal environmental exposures, and bioinformatics. I will obtain extensive training in hESC biology, methods for differentiating these cells into neurons, and approaches for studying the effects of toxicants on this process. HESCs are a valuable model of human embryogenesis because they develop into the three primary germ layers and their derivatives such as neurons. Additionally, I will learn state-of-the-art technique for investigating the transcriptome, e.g., RNA- sequencing (RNA-seq). In general, I theorize that specific environmental exposures during pregnancy impact hESC self-renewal and/or one or more steps in their differentiation to neurons, resulting in detrimental developmental and neurobehavioral outcomes. Here, I aim to test the specific hypothesis that, at environmentally relevant levels, polybrominated diphenyl ethers (PBDE), widely-used flame retardants and potential neurodevelopmental toxicants, disrupt human neurogenesis. First, using an hESC model of neuronal differentiation, I will establish the relevant effective doses of congeners PBDE-47 and/or PBDE-99 (Aim 1). Second, honing in on the relevant ranges, I will characterize the dose-dependent effects of PBDEs at the level of the transcriptome (Aim 2). Finally, I will determine the functional relevance of PBDE gene targets in terms of hESC self-renewal and/or neuronal development (Aim 3). The proposed experiments are the first investigation of PBDE effects on hESCs and their neuronal derivatives. The expected result is identification of alterations at the level of the transcriptome and a functional analysis of genes whose expression levels change. Subsequently, this work could lead to several new research directions. For example, gene-environment interactions identified in vitro could be validated, in terms of adverse neurodevelopmental outcomes, using in vivo models. Additionally, it will be interesting to learn if the PBDE targets are specific to these chemicals or more broadly indicative of neurodevelopmental toxicity. My primary mentor and co-mentors, in complementary fields, will guide me in learning the techniques required to carry out the proposed research. By completing this project, I will master powerful approaches that will enable me to reach my ultimate goal of studying, as an independent investigator, the effects of potential toxicants on seminal aspects of early human development.

描述（由候选人提供）：我寻求资金来扩展我在环境健康科学和发育毒理学方面的培训，通过在三个领域获得其他专业知识：人类胚胎干细胞（HESC）生物学，分子生物学和生物信息学。在这三个领域中使用的基于实验室的方法和统计方法将为我提供过渡到独立学术职业所需的技能，研究环境暴露如何改变早期的人类胚胎发生及其对童年和成人疾病的潜在贡献。对于K99阶段，我的主要导师是苏珊·费舍尔（Susan Fisher）博士，他的专业知识包括使用hESC模型来研究人类发展。我还将收到其他导师的宝贵意见，这些导师是人类神经元发展，产前环境暴露和生物信息学领域的领导者。我将获得有关HESC生物学的广泛培训，将这些细胞区分为神经元的方法以及研究毒物对这一过程的影响的方法。 hESC是人类胚胎发生的宝贵模型，因为它们会发展为三个主要的细菌层及其衍生物，例如神经元。此外，我将学习研究转录组的最新技术，例如RNA测序（RNA-Seq）。通常，我认为怀孕期间的特定环境暴露会影响hESC自我更新和/或一个或多个步骤，从而导致其神经元的分化，从而导致有害的发育和神经行为结果。在这里，我旨在检验以下特定假设，即在环境相关的水平上，多溴二苯基醚（PBDE），广泛使用的阻燃剂和潜在的神经发育有毒物质，破坏了人类神经发生。首先，使用神经元分化的hESC模型，我将建立相关的有效剂量PBDE-47和/或PBDE-99（AIM 1）。其次，在相关范围内磨练，我将表征PBDE在转录组级别上的剂量依赖性作用（AIM 2）。最后，我将根据hESC自我更新和/或神经元发展来确定PBDE基因靶标的功能相关性（AIM 3）。提出的实验是对PBDE对HESC及其神经元衍生物的影响的首次研究。预期的结果是鉴定转录组水平的变化以及表达水平变化的基因的功能分析。随后，这项工作可能会导致几个新的研究方向。例如，使用体内模型，可以在不良神经发育结果中验证在体外鉴定的基因环境相互作用。此外，了解PBDE靶标是否特定于这些化学物质或更广泛地表明神经发育毒性的特异性会很有趣。我在互补领域的主要导师和联合主管将指导我学习进行拟议的研究所需的技术。通过完成该项目，我将掌握强大的方法，使我能够实现作为独立研究者学习潜在毒物对早期人类发展方面的影响的最终目标。