Mechanisms of Environmental-Mixture Induced Metabolic Disruption

项目摘要/摘要 最近的研究表明，通常与肥胖有关的因素，如活动水平， 基因和饮食不足以解释日益恶化的肥胖症流行的规模和速度。 评估其他致病因素的研究越来越多地集中在具有 已证明会影响新陈代谢健康。许多化学物质已被证明能促进 来自多能前体的脂肪细胞承诺或促进甘油三酯蓄积和/或前脂肪细胞 体外增殖，而一些已被进一步证明直接增加体重和/或 体内代谢功能障碍。烷基酚和醇聚乙氧基表面活性剂在 非常规石油和天然气废水、室内粉尘提取物和废水排放，并已 最近被证明能有效地促进脂肪细胞的聚集和前脂肪细胞的增殖 3T3-L1小鼠前脂肪细胞模型。有趣的是，他们似乎通过一种 除了过氧化物酶体增殖物激活受体-γ(PPARγ)以外的机制，通常被认为是 掌握脂肪细胞发育的调节器。这项提议旨在探讨体内代谢的可能性。 这些无处不在的环境污染物通过指导培训目标(K99阶段)进行破坏。这 AIM将为斑马鱼作为模式生物在体外和哺乳动物之间架起桥梁提供重要的经验 活体研究，获得定向分子讯问技术的技能，并最终在一个 评价部分烷基酚和醇聚氧乙烯醚对代谢影响的暴露实验 斑马鱼的健康。此奖学金的独立目标(R00阶段)包括比较代谢 利用体外模型系统进行机制询问，比较斑马鱼和人类受体通路 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 不同于体内常见的内分泌模型。此外，该奖学金将使用应用于 以前的NRSA联谊会，利用修饰的、非固定化的受体配体结合域和高 已知含有烷基酚和酒精的复杂环境样品的分辨质谱学 聚氧乙烯：水力压裂废水、室内粉尘和废水流出物。我们会 定量聚乙氧基化物并测定其对其成脂活性的相对贡献 评估环境混合物对环境代谢干扰的相对影响。这些技能 将被证明对我成为一名学术领域的独立健康科学研究员的职业目标至关重要 通过建立一个全面的管道，从新出现的污染物和混合物的体外测试到 评估对人类健康可能产生的影响。再加上高分辨率质谱学的技能 通过NRSA奖学金获得的，我将在评估和描述复杂环境方面处于独特的地位 污染物和新出现的污染物的混合物，从体外到强大的体内模型。

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.