Identification of Rev-Erb alpha/beta nuclear receptor modulators

在哺乳动物中，能量稳态需要许多代谢过程的精心定时编排。内源性昼夜节律定时系统确保代谢酶在昼夜循环的正确时间产生，以匹配生物体的食物供应和燃料需求。脂质、碳水化合物和葡萄糖的平衡以及激素的分泌都受到昼夜节律的控制。改变人类的睡眠-觉醒周期会导致代谢紊乱，包括胰岛素敏感性降低和葡萄糖耐受不良。在外周组织和器官中，基因表达的时序涉及转录因子，包括核心时钟组分和核受体Rev-Erbα和Rev-Erbß。Rev-Erbs直接调节许多核心时钟成分的表达，但也作为脂质平衡、脂肪生成和葡萄糖输出的主要转录调节因子。与其他核受体类似，Rev-Erbs在其配体结合域（lbd）内含有一个内部疏水腔，可用于药物发现。我们之前确定血红素是Rev-Erbs的内源性配体，并表明可逆血红素结合调节其转录活性。最近，一种单分子类rev - erba /ß调节剂被开发出来，并被证明可以显著改变动物的脂质和葡萄糖基因程序。用Rev-Erb调节剂治疗饮食引起的肥胖动物，其脂肪量显著减少，脂质、胆固醇谱和血糖水平均有改善。我们现在寻求识别许多新的Rev- erbb结合分子，包括特异性作用于Rev- Erbα和Rev- erbß受体的调节剂。为了找到这样的调节剂，我们开发并优化了一种新的高通量测定方法，该方法可以有效地识别结合在lbd内或其四级结构的域-域连接内的分子。我们建议使用MLPCN文库和一系列二级/验证性分析以及基因表达研究开展一对高通量筛选活动，以确定最有希望的rev - erba /ß调节剂。总体目标是发现和开发对肥胖、糖尿病和高脂血症有潜在益处的分子。

DESCRIPTION: In mammals, energy homeostasis requires the carefully timed orchestration of numerous metabolic processes. An endogenous circadian timing system ensures that metabolic enzymes are produced at just the right time within the day-night cycle to match the organism's food availability and fuel needs. Lipid, carbohydrate and glucose balance, as well as the secretion of hormones are all recognized to be under circadian control. Altering the sleep-wake cycles in humans causes metabolic disturbances, including reduced insulin sensitivity and glucose intolerance. In peripheral tissues and organs, the timing of gene expression involves transcription factors including the core clock components and the nuclear receptors Rev-Erbα and Rev-Erbß. The Rev-Erbs directly regulate the expression of many of the core clock components, but also act as master transcriptional regulators of lipid balance, adipogenesis, and glucose output. Similar to other nuclear receptors, the Rev-Erbs contain an internal hydrophobic cavity within their ligand binding domains (LBDs) that can be exploited for drug discovery. We previously identified heme as the endogenous ligand for the Rev-Erbs, and showed that reversible heme binding regulates their transcriptional activities. More recently, a single molecular class of Rev-Erbα/ß modulators was developed and shown to significantly alter the lipid and glucose gene programs in animals. Diet-induced obese animals treated with Rev-Erb modulators display remarkably reduced fat mass and improvements in their lipid and cholesterol profiles and plasma glucose levels. We now seek to identify many new classes of Rev-Erb binding molecules, including modulators that act specifically on each of the Rev- Erbα and Rev-Erbß receptors. To find such modulators, we have developed and optimized a novel high- throughput assay that efficiently identifies molecules that bind within the LBDs, or within the domain-domain junctions of their quaternary structures. We propose to carry out a pair of high-throughput screening campaigns using the MLPCN library and a battery of secondary/confirmatory assays and gene expression studies that identify the most promising Rev-Erbα/ß modulators. The overall goal is to find and develop molecules with potential benefits for obesity, diabetes, and hyperlipidemia.