Mechanisms of Environmental-Mixture Induced Metabolic Disruption

项目总结/摘要 最近的研究表明，通常与肥胖有关的因素，如活动水平， 遗传和饮食不足以解释肥胖症流行病恶化的程度和速度。 越来越多的研究，以评估其他致病因素集中在环境污染物， 被证明会影响代谢健康。许多化学品已被证明可以促进 脂肪细胞从多能前体定型或促进甘油三酯积累和/或前脂肪细胞 在体外增殖，而一些已被进一步证明直接增加体重增加和/或 体内代谢功能障碍。烷基酚和醇聚乙氧基化表面活性剂在 非常规石油和天然气废水，室内房屋灰尘提取物和废水流出物，并已被 最近证明，在糖尿病患者中， 3 T3-L1小鼠前脂肪细胞模型。有趣的是，它们似乎是通过一种 过氧化物酶体增殖物激活受体-γ（过氧化物酶体增殖物激活受体γ）以外的机制，通常被认为是 脂肪细胞发育的主要调节因子。该提案旨在询问体内代谢的潜力， 通过有指导的培训目标（K99阶段），消除这些无处不在的环境污染物对环境的破坏。这 aim将提供利用斑马鱼作为模式生物的重要经验，以在体外和哺乳动物中建立桥梁。 体内研究，获得有针对性的分子审讯技术的技能，并最终在一个指导 暴露实验，评估选定的烷基酚和醇聚乙氧基化物对代谢的影响 斑马鱼的健康本研究（R 00阶段）的独立目的包括比较代谢 机制询问，利用体外模型系统比较斑马鱼和人类受体途径 阐明这些污染物可能影响人类健康的机制，以及这些机制如何 不同于普通的内分泌体内模型。此外，该研究金将使用应用于以下方面的技术： 以前的NRSA奖学金，利用修改，非固定化受体配体结合域和高 已知含有烷基酚和醇的复杂环境样品的分辨质谱法 聚乙氧基化物：水力压裂废水、室内灰尘和废水流出物。我们将 定量聚乙氧基化物并确定它们对这些聚乙氧基化物诱导的脂肪形成活性的相对贡献 环境混合物，以评估其对环境代谢破坏的相对影响。这些技能 我的职业目标是成为一名独立的健康科学研究人员， 通过建立从对新出现的污染物和混合物进行体外测试到 评估对人类健康的推定影响。再加上高分辨率质谱技术 获得NRSA奖学金，我将处于独特的地位，以评估和表征复杂的环境 从体外到体内稳健模型的污染物和新兴污染物的混合物。

Project Summary/Abstract Recent research has demonstrated that factors generally associated with obesity such as activity level, genetics, and diet, are insufficient to account for the magnitude and speed of the worsening obesity epidemic. Increasing research to evaluate other causative factors has focused on environmental contaminants that have been demonstrated to impact metabolic health. Many chemicals have been demonstrated to promote adipocyte commitment from multipotent precursors or promote triglyceride accumulation and/or pre-adipocyte proliferation in vitro, while some have been further demonstrated to directly increase weight gain and/or metabolic dysfunction in vivo. Alkylphenol and alcohol polyethoxylated surfactants are found at high levels in unconventional oil and gas wastewater, indoor house dust extracts, and wastewater effluent, and have been recently demonstrated to promote potent and efficacious lipid accumulation and pre-adipocyte proliferation in the 3T3-L1 mouse pre-adipocyte cell model. Interestingly, they appear to exert this activity through a mechanism other than peroxisome proliferator-activated receptor-gamma (PPARγ), often considered the master regulator of fat cell development. This proposal aims to interrogate the potential for in vivo metabolic disruption by these ubiquitous environmental contaminants through a mentored training aim (K99 phase). This aim will provide crucial experience utilizing zebrafish as a model organism to bridge in vitro and mammalian in vivo research, gaining skills in targeted molecular interrogation techniques, and culminating in a mentored exposure experiment assessing the impact of select alkylphenol and alcohol polyethoxylates on metabolic health of zebrafish. The independent aims of this fellowship (R00 phase) include a comparison of metabolic mechanism interrogation, comparing zebrafish and human receptor pathways utilizing in vitro model systems to elucidate mechanisms through which these contaminants might affect human health and how these may differ from those of a common endocrine in vivo model. Further, this fellowship will use technology applied in the previous NRSA fellowship, utilizing modified, non-immobilized receptor ligand binding domains and high resolution mass spectrometry of complex environmental samples known to contain alkylphenol and alcohol polyethoxylates: hydraulic fracturing wastewater, indoor house dust, and wastewater effluent. We will quantitate polyethoxylates and determine their relative contribution to the adipogenic activity induced by these environmental mixtures to assess their relative influence on environmental metabolic disruption. These skills will prove critical to my career goals of becoming an independent health-science researcher in an academic setting by establishing a comprehensive pipeline from in vitro testing of emerging contaminants and mixtures to assessing putative impacts on human health. Coupled with skills with high resolution mass spectrometry gained under the NRSA fellowship, I will be uniquely placed to assess and characterize complex environmental mixtures of contaminants and emerging contaminants from in vitro to robust in vivo models.