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: 10554600
• 负责人: Dongyin Guan
• 金额: $ 14.91万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 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 2024-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10554600
• 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; circadian transcriptome; 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。时间疗法的概念正吸引越来越多的注意力，以改善药物 当在一天中的最佳时间提供药物时，底层 昼夜节律的分子机制包括核心蛋白质的正反馈环和负反馈环 时钟分子基因在我以前的研究中，由NIDDK的F32博士后培训资助，我们 观察到的昼夜节律增强子和转录组重塑独立于核心蛋白的昼夜节律表达， 饮食诱导肥胖（DIO）小鼠的时钟基因。在我们新的成年肝细胞特异性REV-ERB敲除小鼠中， 模型和限时喂养小鼠，我们也观察到类似的核心时钟基因非依赖性转录组学， 重塑这些观察结果为K 01的应用奠定了基础，并使我们假设， 核心时钟基因在调节昼夜节律增强子活性和基因表达中起着重要作用， 各种正常生理学以及代谢疾病的病理生理学。本提案的目的是 建立昼夜表观基因组重塑和环境挑战的联系，并确定和 表征非核心时钟昼夜节律调节剂，可应用于时间药理学和时间营养学 与代谢性疾病有关的战略。为了实现这一目标，我计划利用无偏见的全基因组 利用转录和生物信息学方法绘制增强子图谱并鉴定调控转录 来自成人肝细胞特异性REV-ERB（Aim 1）的非肝细胞昼夜节律重塑因子（TF）， 在自由进食和限时进食小鼠的肝脏中也是如此（Aim 2）。实际上，我们无偏见的整个基因组- 宽增强子作图和转录组分析揭示了DIO选择性昼夜节律转录因子， 雌激素相关受体γ（ERRγ）。此外，我们无偏见的转录组分析显示， ERRγ在129 S1/SvImJ（129）小鼠肝脏中的表达明显增高。值得注意的是，129只小鼠高度 在B6小鼠中，在产生DIO的饮食中抵抗体重增加和发展代谢功能障碍。作为 在概念验证研究中，我们将确定肝脏ERRγ在对DIO的品系特异性反应中的作用，并扩展 我们的昼夜节律研究的应变特定的背景下（目的3）。 在Mitch Lazar博士的指导下，我加强了对转录调控的训练， 肝代谢在这个K 01奖期间，与主要导师博士拉扎尔和指导我的 作为一个顾问委员会，我将把我对新陈代谢的转录调控的训练扩展到细胞间的串扰水平 （single-nucleus seq），并获得在昼夜生理学中表征非核心时钟基因的经验 和病理生理学。这项培训将使我能够系统地，定量地和功能性地研究分子 参与新陈代谢的联系。我在科学和职业发展方面的进步将受到监督 我的顾问委员会从一个博士后实习生过渡到一个独立的调查员。