Dimensions in Endocrine Disruption: Altering Receptor Coregulator Interactions

描述（由申请人提供）：该项目旨在确定可能的环境内分泌干扰物，其工作机制是配体与核激素受体相互作用后的一种不寻常机制，即直接抑制或增强核激素受体/辅激活因子相互作用本身。这一目标将通过开发时间分辨荧光检测来实现，该检测以稳健和可靠的方式探测核受体与辅激活剂的相互作用。这些检测方法适用于高通量筛选，并将在埃默里大学分子图书馆筛选中心使用，这是一个由NIH资助的路线图研究资源，我们已经获得访问权，以确定可能存在于环境中的可能的辅激活因子结合抑制剂（CBIs）或辅激活因子结合增强剂（CBP）。在化合物被确定为可能的内分泌功能干扰剂或增强剂后，将对其进行二次检测，以确认它们确实通过这些不寻常的CBI或CBP机制发挥作用。这些测定也将使用荧光共振能量转移方法，但在整体组分的浓度中仔细选择改变以阐明机制。最后，基于细胞的试验，如哺乳动物-2-杂交，共转染报告基因试验，或监测的表达反应基因产物将更明确地证明可能的生物或生理效应所产生的暴露于这些化合物。这个项目应该有助于确定核受体信号通路中的一个新的内分泌干扰位点，并确定在这个水平上起作用的化合物。它还应该有助于确定可能存在于环境中或常用产品中的CBI或CBP，这些CBI或CBP可能通过不寻常的机制引起基因组转录模式的改变，这是由于在后配体-受体相互作用水平上操作的核受体调节的信号转导的中断或增强。最终，该项目应有助于识别我们环境中存在的有害化合物，并有望指导采取措施，尽量减少公众对这些物质的接触。 内分泌干扰物是干扰内分泌系统的外源性化合物，改变激素的作用及其向全身发送的信息。该项目旨在确定可能的内分泌干扰物，其作用机制不寻常。大量的化合物将被筛选，以确定那些抑制或增强内分泌作用的结构特征，通过这一不寻常的过程。有了这些信息，就可以采取措施尽量减少人类接触这些新型内分泌干扰物。

DESCRIPTION (provided by applicant): This project aims to identify possible environmental endocrine disrupters working by an unusual mechanism that operates after the interaction of ligands with nuclear hormone receptors, namely, by the direct inhibition or potentiation of the nuclear hormone receptor/coactivator interaction itself. This goal will be accomplished by developing time-resolved fluorescence assays that probe in a robust and reliable manner the interaction of the nuclear receptors with coactivators. These assays are adaptable to high-throughput screening, and will be used at the Emory University Molecular Libraries Screening Center, an NIH-funded Roadmap Research Resource to which we have obtained access, to identify possible coactivator binding inhibitors (CBIs) or coactivator binding potentiators (CBPs) that might be present in the environment. After compounds have been identified as possible disrupters or potentiators of endocrine function, they will be subject to secondary assays to confirm that they indeed act through these unusual CBI or CBP mechanisms. These assays will also use fluorescence resonance energy transfer methodology, but with carefully selected alterations in the concentrations of integral components to elucidate mechanism. Finally, cell-based assays such as mammalian-2-hybrid, cotransfection reporter gene assays, or monitoring of hormone-responsive gene products will more definitively demonstrate the possible biological or physiological effects arising from exposure to these compounds. This project should help pinpoint a novel endocrine disruption site within the nuclear receptor signaling pathway and identify the compounds that work at this level. It should also help identify possible CBIs or CBPs present in the environment or in commonly used products that could be causing alterations in the pattern of genome transcription by an unusual mechanism, due to the interruption or the potentiation of nuclear receptor-regulated signal transduction that operates at a post ligand-receptor interaction level. Ultimately, this project should help in identifying harmful compounds present within our environment and, hopefully, guide measures to minimize exposure of the public to these substances. Endocrine disrupters are exogenous compounds that interfere with the endocrine system, altering hormone action and the messages it sends throughout the body. This project aims to identify possible endocrine disrupters that act by an unusual mechanism. Large numbers of compounds will be screened to determine structural features of those that inhibit or potentiated endocrine action by this unusual process. With this information, steps could be taken to minimize human exposure to these novel types of endocrine disrupters.