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.

项目摘要/摘要 内分泌干扰性化学品(EDCs)包括1,000或更多的合成或自然产生的化学品或 可以干扰荷尔蒙作用的混合物；一些被称为“肥胖症”的混合物已被证明直接 增加体重增加和/或体内脂肪堆积。代谢紊乱的流行，如肥胖， 目前正受到社会的极大关注。据估计，肥胖个人每年为美国贡献2150亿美元 由于医疗保健和生产力丧失而产生的医疗保健成本，还显示出II型糖尿病风险的增加， 心血管疾病、高血压和其他对健康不利的影响。尽管越来越多的关注和 干预措施，美国的发病率仍然很高：9%的婴儿/幼儿，17%的2-19岁，38%的 20岁及以上的成年人都是肥胖者。我们的实验室已经证明，超过一半的房屋灰尘 样本，一个无处不在的EDC暴露源，表现出对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拮抗剂活性(TR-β)。这两种受体 在脂肪生成和甘油三酯积累中起关键作用，以及与这些受体相互作用的内分泌细胞 通常在房屋灰尘的提取物中检测到，这是一种与环境有关的混合物。我们还有 证明了房屋灰尘提取物可以促进甘油三酯在3T3-L1细胞中的积累，这表明 这些受体的破坏可能是导致观察到的活动的原因。然而，许多其他受体可以 有助于脂肪生成，其中一些受体的配体经常在室内被检测到 灰尘。这项建议旨在询问推动甘油三酯在室内积累的主要信号通路。 粉尘提取物通过体外共处理实验和基因表达分析。此外，这一团契将 利用修饰的、非固定化的PPARγ和TRβ的受体配体结合域从 粉尘样本提取物可能与在3T3-L1细胞中观察到的成脂活性有关。这个 建议的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，以及2)提供应用能力 鉴定和定量已知和未知的分析分离和质谱学技术 环境混合物中存在的内源性碳化物。这些技能对于我的近期目标来说是必不可少的 具有分析化学隔离和检测方法的工作知识，可用于识别和 对复杂环境混合物中的致病内分泌干扰物进行量化。此外，这将支持我的长期目标 在学术环境中成为一名独立的健康科学研究员，能够将这些技能与 确定以前未报告的环境内分泌干扰物，并描述其潜在的健康影响。这 在杜克大学一个成功的导师团队的支持下，研究项目将促进我的职业生涯 发展，解决化学品混合物评估中的关键需求，并将有助于更好地开发和评估 识别环境EDCs的工具，以便在未来进行更有针对性的混合物评估。