Hormonal Regulation of Mammalian Gene Expression

描述（由申请人提供）：我们的目标是确定核受体配体和辅因子复合物如何通过控制基因表达模式影响生理和疾病。该提议的基本假设是受体信号传导由配体定向的染色质修饰介导，并且所产生的诱导的表观基因组状态（表观状态）动员基因组或基因网络以产生独特的细胞功能和生理学。为此，在目的I中，我们将使用最先进的蛋白质化学技术来定义在两种选择性调节剂GW 1516和Syndag CBL-28存在下组装的PPARd表观基因组复合物的动态特性。这些将在存在和不存在两种基因表达调节剂-运动模拟AICAR和高脂饮食诱导的肥胖症的情况下进行。具体目标II将建立与上述复合物变化相关的基因表达特征的比较变化。此外，我们将监测受上述配体和调节剂治疗影响的关键代谢参数的变化。最近大规模平行测序技术的可用性和染色质免疫沉淀方法的进步，现在使得有可能在全基因组范围内确定核受体的特定基因组位置（顺式组）。目的III将通过获得ChIP产物的数百万个测序读段并将其映射到参考基因组来确定骨骼肌和培养的肌肉细胞系中的PPARd顺式组。我们还将确定饮食诱导的肥胖症（DIO），受体配体和AICAR如何全面影响PPARd结合。目的IV将通过绘制已用GW 1516、AICAR或高脂饮食处理的细胞和肌肉中的关键组蛋白乙酰化活化和甲基化标记物来定义PPARd的表观遗传特征。在一定程度上，表观基因组控制是一个新的和快速发展的领域，我们认为，核受体在推动它和NR领域向前发展中发挥着关键作用。通过在表观基因组和正常生理学之间建立关键联系，该应用提供了将这种理解扩展到代谢疾病的独特手段，并促进了可以通过治疗基因组来治疗代谢疾病的新药物类别的开发。

DESCRIPTION (provided by applicant): Our goal is to define how nuclear receptor ligands and cofactor complexes influence physiology and disease by controlling patterns of gene expression. The underlying hypothesis of this proposal is that receptor signaling is mediated by ligand directed chromatin modifications and that the resulting induced epigenomic state (epi-state) mobilizes groups or networks of genes to produce unique cell function and physiology. To do this, in Aim I we will use state-of-the-art protein chemistry techniques to define the dynamic properties of PPARd epi-genomic complexes assembled in the presence of two selective modulators GW1516 and Syndag CBL-28. These will be performed in the presence and absence of two gene expression modulators-the exercise mimetic AICAR and high-fat diet induced obesity. Specific Aim II will establish the comparative changes in gene expression signatures that correlate to changes in the above complexes. In addition, we will monitor shifts in key metabolic parameters influenced by treatment with the above ligands and modulators. The recent availability of massively parallel sequencing technology and advances in methods for chromatin immunoprecipitation now makes it possible to determine the specific genomic locations (cistrome) of nuclear receptors on a genome-wide scale. Aim III will determine the PPARd cistrome in skeletal muscle and cultured muscle cell lines by obtaining millions of sequencing reads of ChIP products and mapping these to reference genomes. We will also determine how diet induced obesity (DIO), receptor ligands and AICAR globally impact PPARd binding. Aim IV will define the epigenetic signatures of PPARd by mapping key histone acetylation activation and methylation markers in cells and muscle that have been treated with GW1516, AICAR or high fat diets. To the extent that epigenomic control is a new and rapidly evolving field, we believe that nuclear receptors play a critical role in driving it and the NR field forward. By making key links between the epigenome and normal physiology this application provides a unique means to extend this understanding to metabolic disease and facilitates the development of new classes of drugs that can treat diseases of metabolism by treating the genome.