Interrogation of molecular mechanisms involved in driving adipogenesis in environmental mixtures and novel analytical techniques for identifying putative causative chemicals

探究环境混合物中驱动脂肪生成的分子机制以及用于识别假定致病化学物质的新颖分析技术

• 批准号: 9328723
• 负责人: Christopher Dennis Kassotis
• 金额: $ 5.71万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-05-01 至 2020-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9328723
• 关键词: 19 year old; 3T3-L1 Cells; Address; Adipocytes; Adult; Affinity; Agonist; Androgen Receptor; Animal Model; Attention; Automobile Driving; Award; Biological Assay; Breathing; Cardiovascular Diseases; Caring; Chemical Exposure; Chemical Structure; Chemicals; Child; Competence; Complex; Complex Mixtures; Data; Detection; Development; Disease; Dust; Endocrine Disruptors; Estrogen Receptors; Evaluation; Exhibits; Exposure to; Fellowship; Fostering; Future; Gene Expression; Gene Expression Profiling; Genes; Glucocorticoid Receptor; Goals; Health; Health Care Costs; Health Sciences; Health trends; Hormonal; Hormones; House Dust; Hypertension; In Vitro; Individual; Infant; Intervention; Knowledge; Laboratories; Life; Ligand Binding Domain; Ligands; Lipids; Liver X Receptor; Mass Spectrum Analysis; Medical; Mentorship; Metabolic Diseases; Metabolism; Methods; Molecular; Mus; National Research Service Awards; Non-Insulin-Dependent Diabetes Mellitus; Nuclear Receptors; Obesity; Oral; PPAR gamma; Pathway interactions; Play; Positioning Attribute; Prevalence; Productivity; RXR; Receptor Signaling; Regulation; Reporting; Research; Research Personnel; Research Proposals; Research Training; Resolution; Risk; Role; Route; Sampling; Signal Pathway; Signal Transduction; Source; Techniques; Testing; Thyroid Gland; Time; Toddler; Training; Triglycerides; Universities; Weight Gain; Work; adipocyte differentiation; age group; aged; career development; cost; critical period; environmental justice; experimental study; exposed human population; in vivo; indoor exposure; lipid biosynthesis; new technology; novel; obesogen; programs; public health relevance; receptor; response; skills; tool

Project Summary/Abstract Endocrine disrupting chemicals (EDCs) include 1,000 or more synthetic or naturally occurring chemicals or mixtures that can interfere with hormone action; some, termed “obesogens”, have been shown to directly increase weight gain and/or lipid accumulation in vivo. The prevalence of metabolic disorders, such as obesity, is currently of great societal concern. Obese individuals contribute to an estimated $215 billion in annual US health care costs due to medical care and lost productivity, and also exhibit increased risks of type II diabetes, cardiovascular disease, hypertension, and other adverse health effects. Despite increased attention and interventions, rates of US occurrence remain high: 9% of infants/toddlers, 17% of 2-19 year olds, and 38% of adults aged 20 and older are obese. Our laboratory has demonstrated that more than half of house dust samples, a ubiquitous EDC exposure source, exhibit agonist activity for peroxisome proliferator-activated receptor-gamma (PPARγ) and/or antagonist activity for thyroid receptor-beta (TRβ). Both of these receptors play critical roles in adipogenesis and triglyceride accumulation, and EDCs that interact with these receptors are commonly detected in extracts of house dust, an environmentally relevant mixture. We have also demonstrated that house dust extracts can promote triglyceride accumulation in 3T3-L1 cells, suggesting that disruption of these receptors may contribute to the observed activity. However, many other receptors can contribute to adipogenesis, and ligands for some of these receptors have been frequently detected in house dust. This proposal aims to interrogate the main signaling pathways driving triglyceride accumulation in house dust extracts via in vitro co-treatment experiments and gene expression analyses. Further, this fellowship will utilize modified, non-immobilized receptor ligand binding domains for PPARγ and TRβ to isolate ligands from dust sample extracts that may be responsible for the adipogenic activity observed in 3T3-L1 cells. The proposed NRSA postdoctoral fellowship will 1) provide additional training in molecular mechanism characterization through gene expression and mixture analyses and 2) provide competency in applying analytical isolation and mass spectrometry techniques to identify and quantitate both known and unknown EDCs present in environmental mixtures. These skills are essential to my immediate goals of developing a working knowledge of analytical chemical isolation and detection methods that can be used to identify and quantify causative EDCs in complex environmental mixtures. Furthermore, this will support my long-term goals of becoming an independent health-science researcher in an academic setting able to couple these skills to identify previously unreported environmental EDCs and characterize their potential health impacts. This research program, supported by a successful mentorship team at Duke University, will foster my career development, address critical needs in chemical mixture assessments, and will help develop and assess better tools to identify environmental EDCs in order to allow for more directed mixture assessments in the future.

项目概要/摘要 内分泌干​​扰化学物质 (EDC) 包括 1,000 种或更多合成或天然存在的化学物质，或 可以干扰激素作用的混合物；一些被称为“致肥胖物质”的混合物已被证明可以直接干扰激素作用。 增加体内体重增加和/或脂质积累。代谢紊乱的患病率，例如肥胖， 目前，肥胖者每年为美国带来约 2150 亿美元的收入，受到社会极大关注。 由于医疗保健和生产力损失导致的医疗保健费用，并且还表现出 II 型糖尿病的风险增加， 尽管人们越来越关注和关注心血管疾病、高血压和其他不良健康影响。 尽管采取了干预措施，美国的发生率仍然很高：婴儿/幼儿为 9%，2-19 岁儿童为 17%，2-19 岁儿童为 38% 我们的实验室证明，超过一半的室内灰尘导致 20 岁及以上的成年人肥胖。 样品是一种普遍存在的 EDC 暴露源，表现出过氧化物酶体增殖物激活的激动剂活性 受体-γ (PPARγ) 和/或甲状腺受体-β (TRβ) 的拮抗剂活性。 在脂肪生成和甘油三酯积累中发挥关键作用，并且 EDC 与这些受体相互作用 通常在房屋灰尘的提取物中检测到，这是一种与环境相关的混合物。 证明室内灰尘提取物可以促进 3T3-L1 细胞中甘油三酯的积累，这表明 这些受体的破坏可能有助于观察到的活性，但是，许多其他受体也可以。 有助于脂肪生成，其中一些受体的配体已在内部经常检测到 该提案旨在探究驱动室内甘油三酯积累的主要信号通路。 此外，该奖学金还将通过体外共处理实验和基因表达分析来研究灰尘提取物。 利用 PPARγ 和 TRβ 的修饰的非固定化受体配体结合域来分离配体 灰尘样品提取物可能与 3T3-L1 细胞中观察到的脂肪形成活性有关。 拟议的 NRSA 博士后奖学金将 1) 提供分子机制方面的额外培训 通过基因表达和混合物分析进行表征，2）提供应用能力 用于识别和定量已知和未知物质的分析分离和质谱技术 环境混合物中存在 EDC，这些技能对于我开发 EDC 的近期目标至关重要。 分析化学分离和检测方法的工作知识，可用于识别和 此外，这将支持我的长期目标。 成为学术环境中的独立健康科学研究人员，能够将这些技能与 识别以前未报告的环境 EDC 并描述其潜在的健康影响。 由杜克大学成功的导师团队支持的研究计划将促进我的职业生涯 开发，解决化学混合物评估的关键需求，并将有助于更好地开发和评估 识别环境 EDC 的工具，以便将来进行更有针对性的混合物评估。