Defining the role of non-clock genes in circadian physiology and pathophysiology

定义非时钟基因在昼夜节律生理学和病理生理学中的作用

• 批准号: 10224846
• 负责人: Dongyin Guan
• 金额: $ 13.1万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-08-01 至 2021-08-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10224846
• 关键词: 129 Mouse; Adopted; Adult; Advisory Committees; Affect; Attention; Bile Acid Biosynthesis Pathway; Binding; Bioinformatics; Biology; Cardiovascular Diseases; Cell Nucleus; Cells; Chronotherapy; Circadian Rhythms; Circadian desynchrony; Data; Diabetes Mellitus; Diet; Disease; Drug Delivery Systems; Drug toxicity; Endothelial Cells; Enhancers; Environmental Risk Factor; Epigenetic Process; Feedback; Food; Foundations; Functional disorder; Gene Expression; Genes; Genetic; Genetic Transcription; Goals; Grant; Hepatic; Hepatocyte; Homeostasis; Hypertension; Individual; Isotopes; Knockout Mice; Knowledge; Kupffer Cells; Lead; Light; Lipids; Liver; Malignant Neoplasms; Mammals; Maps; Mentored Research Scientist Development Award; Mentorship; Metabolic; Metabolic Diseases; Metabolic dysfunction; Metabolism; Methods; Molecular; Mus; National Institute of Diabetes and Digestive and Kidney Diseases; Neurons; Non-Insulin-Dependent Diabetes Mellitus; Obese Mice; Overnutrition; PPAR alpha; Pathway interactions; Periodicity; Peripheral; Pharmaceutical Preparations; Pharmacology; Phase; Physiological; Physiology; Play; Process; Regulation; Research; Research Personnel; Resistance; Risk Factors; Role; Science; Sleep Disorders; Supervision; Testing; Therapeutic; Time; Time-restricted feeding; Tissues; Training; Transcriptional Regulation; Weight Gain; base; career; career development; circadian; circadian pacemaker; circadian regulation; clinical application; diet-induced obesity; drug efficacy; environmental change; epidemiology study; epigenomics; estrogen-related receptor; experience; feeding; genome-wide; improved; in vivo; lipid metabolism; liver metabolism; metabolic abnormality assessment; molecular clock; mouse model; novel; nutrition; obesity risk; post-doctoral training; response; shift work; skills; suprachiasmatic nucleus; transcription factor; transcriptome; transcriptomics; whole genome

Project Abstract Circadian misalignment is a risk factor for many diseases, such as type-2 diabetes, cardiovascular disease, hypertension, and cancer. The concept of chronotherapy is attracting more and more attention to improving drug efficacy and diminishing drug toxicity when drugs are provided at the optimized time of the day. The underlying molecular mechanism of circadian rhythm includes the interlocking positive and negative feedback loops of core clock molecular genes. In my previous study, supported by an F32 post-doctoral training grant from NIDDK, we observed circadian enhancer and transcriptome remodeling is independent of the circadian expression of core clock genes in diet-induced obesity (DIO) mice. In our new adult hepatocyte-specific REV-ERB knockout mice model and time-restricted feeding mouse, we also observed similar core clock gene-independent transcriptomic remodeling. These observations set a foundation of this K01 application and lead us to hypothesize that non- core clock genes play an essential role in the regulation of circadian enhancer activity and gene expression in various normal physiology as well as the pathophysiology of metabolic diseases. The goal of this proposal is to establish the connection of circadian epigenomic remodeling and environmental challenge and to identify and characterize non-core clock circadian regulators that can apply to chrono-pharmacological and chrono-nutritive strategies relating to metabolic diseases. To accomplish this goal, I plan to utilize unbiased genome-wide transcriptional and bioinformatics methods to map enhancer landscape and identify regulatory transcription factors (TFs) for circadian remodeling in non-hepatocytes from adult hepatocyte-specific REV-ERB (Aim 1) and also in livers of ad libitum feeding and time-restricted feeding mice (Aim 2). Actually, our unbiased whole genome- wide enhancer mapping and transcriptome analysis revealed a DIO-selective circadian transcription factor, Estrogen Related Receptor Gamma (ERRγ). Moreover, our unbiased transcriptome profiling revealed that the expression of ERRγ is markedly higher in the livers of 129S1/SvImJ (129) mice. Of note, 129 mice are highly resistant to gaining weight and developing metabolic dysfunction on diets that produce DIO in B6 mice. As a proof-of-concept study, we will determine the role of hepatic ERRγ in strain-specific response to DIO and extend our circadian rhythm study to a strain-specific context (Aim 3). Under the mentorship of Dr. Mitch Lazar, I have strengthened my training in the transcriptional regulation of hepatic metabolism. In this K01 award period, with primary mentorship from Dr. Lazar and the guidance of my advisory committee, I will extend my training on transcription regulation of metabolism to cell-cell crosstalk level (single-nuclei seq) and gain experiences about characterization of non-core clock genes in circadian physiology and pathophysiology. This training will allow me to systemically, quantitatively and functionally study molecular connections involved in metabolism. My progress in science and career development will be under supervision of my advisory committee for the transition from a postdoctoral trainee to an independent investigator.

项目摘要 昼夜节律失调是许多疾病的危险因素，例如2型糖尿病，心血管疾病， 高血压和癌症。年代疗法的概念吸引了越来越关注改善药物 当在一天中的优化时间提供药物时，功效和药物毒性降低。基础 昼夜节律的分子机制包括核心的正面和负反馈回路 时钟分子基因。在我以前的研究中，在NIDDK的F32博士后培训补助金的支持下，我们 观察到的昼夜节律增强子和转录组重塑与核心的昼夜节律无关 饮食诱导的肥胖症（DIO）小鼠中的时钟基因。在我们的新成年肝细胞特异性转速淘汰小鼠中 模型和时间限制的喂食小鼠，我们还观察到了相似的核心时钟基因独立的转录组 重塑。这些观察结果为这一K01应用奠定了基础，并使我们假设非 - 核心时钟基因在调节昼夜节律增强剂活性和基因表达中起着至关重要的作用 各种正常生理以及代谢疾病的病理生理学。该提议的目的是 建立昼夜节学表观基因组重塑和环境挑战的联系，并识别和 表征非核钟昼夜节律调节器，可以适用于计时剂和计时效果 与代谢疾病有关的策略。为了实现这一目标，我计划利用无偏基因组的公正 转录和生物信息学方法绘制增强剂景观并确定调节转录 成人肝细胞特异性REV-ERB（AIM 1）和 同样在多次喂养和时间限制的喂养小鼠的肝脏中（AIM 2）。实际上，我们公正的整个基因组 - 广泛的增强器映射和转录组分析显示，二氧化合物选择性的昼夜节律转录因子， 雌激素相关的受体伽马（ERRγ）。此外，我们的公正转录组分析表明 在129S1/SVIMJ（129）小鼠的生命中，ERRγ的表达明显更高。值得注意的是，129只小鼠高度 对在B6小鼠中产生DIO的饮食上的体重和代谢功能障碍有抵抗力。作为 概念验证研究，我们将确定肝素ERRγ在特异性反应中的作用，并扩展 我们的昼夜节律研究对特定于菌株的背景（AIM 3）。 在米奇·拉扎尔（Mitch Lazar）博士的心态下，我加强了我的培训 肝代谢。在这个K01奖励期间，Lazar博士的主要精神训练和我的指导 咨询委员会，我将把有关代谢的转录调节培训扩展到细胞细胞串扰级别 （单核Seq）并获得有关昼夜节律生理中非核心时钟基因表征的经验 和病理生理学。这种训练将使我能够系统地，定量和功能研究分子 新陈代谢涉及的联系。我在科学和职业发展方面的进步将受到监督 我的咨询委员会从博士后实习生到独立调查员的过渡。