Estrogen receptor control of inflammatory gene expression

雌激素受体控制炎症基因表达

• 批准号: 9515944
• 负责人: Kendall W Nettles
• 金额: $ 43.2万
• 依托单位: SCRIPPS FLORIDA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-01 至 2020-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9515944
• 关键词: Agonist; Autoimmunity; BIRC3 gene; Binding; Biological Assay; Biology; Cardiovascular Diseases; Categories; Chemical Structure; Chemicals; Chemistry; Communicable Diseases; Complex; Crystallization; DNA; Diabetes Mellitus; Disease; Endocrine; Epigenetic Process; Estrogen Receptor alpha; Estrogen Receptors; Estrogens; Gene Expression; Genes; Genetic Transcription; Genomic approach; Goals; Group Structure; Hormones; IL6 gene; IL8 gene; Impaired cognition; Impairment; Inflammation; Inflammatory; Inflammatory Response; Libraries; Ligand Binding Domain; Ligands; Linear Regressions; Manuals; Mediating; Menopause; Metabolic Diseases; Molecular; Molecular Conformation; Mutagenesis; Nerve Degeneration; Nuclear Receptors; Obesity; Outcome; Pathway interactions; Phenotype; Physiological; Physiology; Principal Component Analysis; Publishing; Regulation; Response Elements; Signal Pathway; Signal Transduction; Site; Specificity; Statistical Data Interpretation; Statistical Models; Structure; System; Systems Biology; TNF gene; Techniques; Time; Tissues; Work; X-Ray Crystallography; bone; causal model; conformer; deletion analysis; drug discovery; estrogenic; functional genomics; gender difference; human disease; receptor; recruit; scaffold; screening; small hairpin RNA; small molecule; structural biology; therapeutic target; tool

Menopause induces systemic inflammation that contributes to impaired endocrine and bone function, metabolic disease, obesity, as well as autoimmunity and cognitive decline. These observations highlight a critical need to understand the relationship between estrogen signaling and inflammation. The overarching problem in both our basic understanding and therapeutic targeting of nuclear receptors remains an almost complete lack of understanding of how small differences in ligands can drive widely different transcriptional outcomes, including pathway and tissue-selective signaling. The goal is to define the chemical, structural, and molecular rules for how the estrogen receptor-α (ERα) ligands control inflammatory gene expression through binding directly or indirectly to DNA and altering coregulator recruitment, which in turn controls gene expression. A systems biology approach to chemical biology and structural biology will be used to reveal rules for how ligands achieve signaling specificity. Specific Aim 1. Discover and characterize the epigenetic regulome that mediates differential effects of ERα ligands on the inflammatory response. Functional genomics approaches will be used to identify three sets of inflammatory gene sets that we hypothesize will show distinct rules for gene control: 1) TNF-induced, E2 modulated genes with composite ERE/κB sites 2) TNF-induced, E2 modulated genes with ERα tethering to κB response elements; and 3) TNF-induce genes where ERα bounds but does not modulate. We will pick 3-4 representative genes from each category to target with an shRNA screen targeting all coregulators (~1200 shRNAmir targeting ~300 genes) to identify those required for ERα modulation. Coregulator-ERα interaction screens will be built and profiled with a compound library of 500 ERα ligands. Statistical models will enable identification of specific coregulator interactions that predict the inflammatory response to different classes of ligands and ties them to specific structural perturbations. Specific Aim 2. Identify the chemical and structural rules by which ERα transduces chemical structure into recruitment of specific signaling complexes that control inflammatory gene expression. Analysis of >100 crystal structures will be used to compute all atom distance matrices and use principal component analysis and multiple linear regression to identify ligand-induced perturbations that predict (1) regulation of specific genes, and (2) interaction with specific coregulators defined in Specific Aim 1. Impact: This work will identify structural rules for how ERα ligands differentially regulate inflammatory gene expression through the ligand-receptor interface allosterically controlling recruitment of an ensemble of specific coregulatory complexes. Identifying causal models for how ligands achieve signaling specificity impacts drug discovery and our understanding of the physiology underlying a variety of hormone-sensitive physiological systems and disease states, and should be readily translatable to other allosteric signaling systems including other nuclear receptors and GPCRs.

更年期引起全身炎症，导致内分泌和骨功能受损， 代谢疾病、肥胖症以及自身免疫和认知能力下降。这些观察突出了一个 迫切需要了解雌激素信号和炎症之间的关系。总体 在我们对核受体的基本理解和治疗靶向方面， 完全缺乏对配体的微小差异如何驱动广泛不同的转录的理解， 结果，包括通路和组织选择性信号传导。目标是确定化学、结构和 雌激素受体-α（ERα）配体如何通过以下途径控制炎症基因表达的分子规则 直接或间接与DNA结合，改变辅调节因子的募集，进而控制基因 表情一个系统生物学的方法，化学生物学和结构生物学将被用来揭示规则 配体如何实现信号特异性。具体目标1.发现和表征表观遗传 介导ERα配体对炎症反应的不同作用的调节基因。功能 基因组学方法将被用于确定三组炎症基因组，我们假设这些基因组将 显示了不同的基因控制规则：1）TNF诱导的，E2调节的具有复合ERE/κB位点的基因2） TNF诱导的E2调节基因，ERα与κB反应元件相连; 3）TNF诱导的基因 其中ERα结合但不调节。我们将从每个类别中挑选3-4个代表性基因进行靶向 用靶向所有辅调节子的shRNA筛选（靶向约300个基因的约1200个shRNAmir）来鉴定那些 ERα调节所需的。将构建辅调节因子-ER α相互作用筛选，并使用化合物进行分析 500种ERα配体的文库。统计模型将能够识别特定的共调节因子相互作用， 预测对不同类型配体的炎症反应，并将它们与特定的结构联系起来， 扰动具体目标2。确定ERα转导的化学和结构规则 控制炎症基因的特定信号复合物的募集 表情对>100种晶体结构的分析将用于计算所有原子距离矩阵，并使用 主成分分析和多元线性回归来识别配体诱导的扰动， (1)特定基因的调节，和（2）与特定目标1中定义的特定辅助调节因子的相互作用。影响： 这项工作将确定ERα配体如何差异调节炎症基因表达的结构规则 通过配体-受体界面变构地控制特异性受体的集合的募集， 共调节复合物识别配体如何实现信号特异性影响药物的因果模型 发现和我们对生理学的理解，这些生理学基础是各种对疼痛敏感的生理学， 系统和疾病状态，并且应该容易地翻译到其他变构信号传导系统，包括 其他核受体和GPCR。