The Role of Cryptochromes in Circadian Regulation of Metabolism

隐花色素在代谢昼夜节律调节中的作用

• 批准号: 9175163
• 负责人: STEVE A KAY
• 金额: $ 69.14万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9175163
• 关键词: Affinity; Alleles; Architecture; Binding; Binding Sites; Biochemical; Biochemistry; Bioinformatics; Biological Assay; Biological Clocks; Biological Models; Biological Process; Biology; Cardiovascular Diseases; Cell Nucleus; Cells; ChIP-seq; Chemicals; Chromatin; Circadian Rhythms; Clinical; Clock protein; Collaborations; Collection; Complex; Cytoplasm; DNA Binding; DNA-Protein Interaction; Data; Data Set; Diabetes Mellitus; Disease; Dose; Enhancers; Gene Targeting; Genes; Genetic; Genomics; Gluconeogenesis; Health; Hepatocyte; Hot Spot; Hour; Human; Impairment; In Vitro; Jet Lag Syndrome; Length; Liver; Locales; Mediating; Metabolic; Metabolic Diseases; Metabolic Pathway; Metabolism; Modeling; Molecular; Molecular Target; Mus; Nuclear; Nucleic Acid Regulatory Sequences; Organism; Pathway interactions; Periodicity; Physiological Processes; Physiology; Proteins; Proteomics; Reagent; Regulation; Repression; Resolution; Roentgen Rays; Role; Series; Sleep Wake Cycle; Sleeplessness; Specificity; Structure; System; Techniques; Therapeutic; Time; Tissues; Translations; Validation; Work; X-Ray Crystallography; abstracting; base; blood glucose regulation; chromosome conformation capture; circadian pacemaker; cryptochrome; design; gene repression; genome-wide; glucose tolerance; improved; in vivo; insight; mutant; next generation; novel; novel therapeutics; protein metabolism; small molecule; success; tool; transcription factor; transcriptome sequencing; transcriptomics

Project Summary / Abstract Circadian rhythms are pervasive among organisms, allowing them to anticipate and adapt to the predictable 24-hour day-night cycle. Their function is to temporally coordinate physiological processes, such as metabolism, within the organism. Consequently, the disruption of circadian rhythms leads to desynchronized internal clocks and complex metabolic disorders, such as diabetes. The mechanism of how the clock controls downstream metabolic pathways is not well-established. One of the central players in this relationship is the core clock gene Cryptochrome (Cry). CRY is necessary to maintain rhythmicity and determine period length, but it has also been implicated in diabetes and glucose tolerance. Until recently, CRY was thought to function only in the nucleus; our recent unanticipated findings indicate it also inhibits gluconeogenesis in the cytoplasm through its interaction with Gsα. In parallel, nuclear CRY also regulates gluconeogenesis, albeit through a completely different pathway. Together, this two-pronged approach allows CRY to fine-tune its regulation of glucose homeostasis; however, its presence in two subcellular locales has made it difficult to study its compartment-specific mechanisms. To overcome this challenge, we created two unique reagents that localize CRY to each region. Cytosolic CRY will be studied at the atomic and cellular level to identify its binding partners and how their interactions determine their biochemical functions. Nuclear CRY will be investigated on the genomic scale to uncover its interactions with other transcription factors in the enhancers of gluconeogenesis genes. Chromosome conformation capture techniques will enable us to model nucleus-wide hepatocyte-specific enhancer architecture. On the therapeutic front, we will characterize the mechanism of action of novel clock-modifying chemical compounds identified from our screens. These compounds have the potential to identify novel clock genes and to regulate metabolism, paving the way towards clinical translation. The use of these techniques to study how CRY controls gluconeogenesis will be a proof-of-concept for how the clock achieves precision in modulating a tissue-specific metabolic pathway. The success of these studies will significantly improve the understanding of the crosstalk between the biological clock and physiology or disease states, as well as provide a proof-of-concept model for applying cell-based findings in improving human health.

项目摘要 /摘要 昼夜节律在生物体中普遍存在，使它们能够预期并适应可预测的 24小时昼夜周期。它们的功能是临时协调物理过程，例如 代谢，在生物体中。因此，昼夜节律的破坏导致了异议 内部时钟和复杂的代谢性疾病，例如糖尿病。时钟如何控制的机制 下游的代谢途径没有建立良好的。这种关系中的核心参与者之一是 核心时钟基因加密色素（哭泣）。哭泣是维持节奏性并确定周期长度的必要条件 但是它也暗示在糖尿病和葡萄糖耐受性中。直到最近，哭泣仍被认为起作用 仅在核中；我们最近的意外发现，表明它也抑制了细胞质中的谷杆菌生成 通过与GSα的相互作用。同时，核哭泣还调节糖异生，尽管通过 完全不同的途径。这种两管沟的方法一起允许哭泣，以微调其调节 葡萄糖稳态；但是，它在两个亚细胞的存在中的存在使得很难研究其 隔室特异性机制。为了克服这一挑战，我们创建了两个本地化的独特试剂 向每个区域哭泣。胞质哭泣将在原子和细胞水平上进行研究，以识别其结合 伙伴以及他们的相互作用如何决定其生化功能。将调查核哭泣 在增强子中发现其与其他转录因子相互作用的基因组量表 糖异生基因。染色体会议捕获技术将使我们能够模拟整个核的模型 肝细胞特异性增强剂体系结构。在治疗方面，我们将表征 从我们的屏幕上鉴定出的新型时钟修改化合物的作用。这些化合物具有 潜力鉴定新的时钟基因并调节新陈代谢，为临床翻译铺平道路。 使用这些技术来研究哭泣如何控制葡萄糖的方法将是概念概念的证明 时钟在调节组织特异性代谢途径时达到了精度。这些研究的成功将 显着提高对生物钟与生理或疾病之间串扰的理解 国家，以及提供概念验证模型，用于在改善人类健康中应用基于细胞的发现。