Role of Ror proteins in the mammalian circadian clock

Ror 蛋白在哺乳动物生物钟中的作用

• 批准号: 8656526
• 负责人: STEVE A KAY
• 金额: $ 24.56万
• 依托单位: UNIVERSITY OF SOUTHERN CALIFORNIA
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-01 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8656526
• 关键词: Role; Ror; proteins; mammalian; circadian

DESCRIPTION (provided by applicant): Circadian rhythms exist in almost all organisms to anticipate the daily changes in the environment and to temporally coordinate biological processes. Although much progress has been made to identify the components of the mammalian clock and the mechanisms by which it controls physiology, the knowledge base is by no means complete. We propose to further elucidate the architecture of the liver clock and its relationship with metabolism by generating and integrating divergent types of large-scale data, followed by validation of these data in vivo. In order to isolate and study the cell-autonomous clock, we have identified a unique cell line, Met Murine Hepatocyte-Day 3 (MMH-D3) that retains many metabolic functions and exhibits robust circadian rhythms. Thus we will be able to interrogate both the clock and metabolic pathways and study their interactions. We have already obtained transcriptomic and metabolomic data for MMH-D3 time-courses (every 2-hours for 48-hours) to identify cycling entities. Furthermore, we will perform chromatin immunoprecipitation followed by sequencing (ChIP-seq) for REV-ERB1, a nuclear hormone receptor that is part of the clock and a regulator of lipid metabolism, to explore one of the mechanisms of how the clock controls metabolism. To identify novel proteins that act as systemic cues for the liver clock, we will perform high- throughput screening of an invaluable library of over 6,000 secreted proteins. To our knowledge, this is one of the broadest and diverse data sets collected for the analysis of the circadian clock. To generate the most comprehensive model of the clock, these data sets will not only be evaluated individually, but will be integrated to reconstruct biological networks using methods based on vector auto-regression. Such networks have the ability to predict regulation and interaction within and between the different layers, as well as their associations with metabolic pathways. The hypotheses generated by the network model will be verified using adenovirus techniques in vivo, which will also enable us to differentiate between cell-autonomous and systemic signals governing the liver clock. The clock clearly has an influence on proper physiological function, thus a better understanding of circadian clockwork will improve our understanding of the mechanisms that underlie human disease; this will refine diagnostic and therapeutic strategies.

描述（由申请人提供）：几乎所有生物体中都存在昼夜节律，以预测环境的日常变化和时间协调生物学过程。尽管已经取得了很大的进步来确定哺乳动物时钟的组成部分及其控制生理学的机制，但知识库绝不是完整的。我们建议通过生成和集成不同类型的大规模数据，然后在体内验证这些数据，以进一步阐明肝时钟的结构及其与代谢的关系。为了隔离和研究细胞自主时钟，我们已经确定了独特的细胞系，遇到了鼠肝细胞3（MMH-D3），该鼠类保留了许多代谢功能，并表现出强大的昼夜节律。因此，我们将能够询问时钟和代谢途径并研究其相互作用。我们已经获得了MMH-D3时间场景（每2小时的48小时）的转录组和代谢组数据，以识别循环实体。此外，我们将执行染色质免疫沉淀，然后进行测序（chip-seq）Rev-erb1，Rev-erb1是一种核激素受体，是时钟的一部分，也是脂质代谢的调节剂，以探索如何控制时钟的机制之一。为了确定充当肝时钟系统提示的新型蛋白质，我们将对超过6,000种分泌蛋白的宝贵库进行高吞吐量筛选。据我们所知，这是用于分析昼夜节律时钟的最广泛，最多样化的数据集之一。为了生成最全面的时钟模型，这些数据集将不仅可以单独评估，而且将使用基于矢量自动回归的方法集成到重建生物网络。这样的网络具有预测不同层内和之间之间的调节和相互作用及其与代谢途径的关联。网络模型生成的假设将使用体内腺病毒技术进行验证，这也将使我们能够区分控制肝时钟的细胞自主和全身信号。时钟显然对适当的生理功能有影响，因此对昼夜节律的更好理解将提高我们对构成人类疾病基础机制的理解。这将完善诊断和治疗策略。