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​​扰化学物质（EDC）包括1,000或更多的合成或天然化学物质或 可以干扰骑马动作的混合物；有些被称为“肥胖症”，已被证明直接 增加体内体重增加和/或脂质积累。肥胖症等代谢疾病的患病率， 目前非常关注社会问题。肥胖个人为美国年度估计贡献了2150亿美元 医疗保健费用由于医疗保健和生产力失去，还暴露了II型糖尿病风险的增加， 心血管疾病，高血压和其他不良健康影响。尽管注意力越来越大 干预措施，美国发生率仍然很高：婴儿/幼儿的9％，2-19岁的17％和38％ 20岁及20岁以上的成年人肥胖。我们的实验室证明了超过一半的房屋灰尘 样品是一种无处不在的EDC暴露来源，暴露于过氧化物组增殖物激活的激动剂活性 甲状腺受体β（TRβ）的受体γ（PPARγ）和/或拮抗剂活性。这两个受体 在脂肪生成和甘油三酸酯积累中起关键作用，与这些受体相互作用的EDC 通常在房屋灰尘的提取物（一种与环境相关的混合物）的提取物中检测到。我们也有 证明房屋灰尘提取物可以促进3T3-L1细胞中的甘油三酸酯的积累，这表明 这些受体的破坏可能有助于观察到的活性。但是，许多其他受体可以 在房屋中经常检测到这些受体中的某些受体的脂肪形成和配体 灰尘。该提案旨在询问驱动在房屋中触发积累的主要信号通路 通过体外共同处理实验和基因表达分析进行粉尘提取。此外，这个奖学金将 PPARγ和TRβ的经过修改的，非弹性的受体配体结合结构域，以分离配体从 可能导致3T3-L1细胞中观察到的掺杂活性的灰尘样品提取物。 拟议的NRSA博士后奖学金将1）提供分子机制的额外培训 通过基因表达和混合分析表征，2）提供了应用的能力 分析隔离和质谱技术，以识别和定量已知和未知 EDC存在于环境混合物中。这些技能对于我开发一个目标至关重要 分析化学隔离和检测方法的工作知识，可用于识别和 在复杂的环境混合物中量化病原体EDC。此外，这将支持我的长期目标 在学术环境中成为独立的健康科学研究人员，能够伴随这些技能 确定以前未报告的环境EDC，并表征其潜在的健康影响。这 在杜克大学成功的Mentalship团队的支持下，研究计划将促进我的职业生涯 开发，满足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，以便将来进行更多的定向混合物评估。