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、 高血压和癌症的概念越来越受到人们的关注。 当在一天中的最佳时间提供药物时，可以提高疗效并减少药物毒性。 昼夜节律的分子机制包括核心的连锁正反馈环和负反馈环 在我之前的研究中，在 NIDDK 的 F32 博士后培训资助的支持下，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表达无关 在我们新的成年肝细胞特异性 REV-ERB 敲除小鼠中，饮食诱导肥胖 (DIO) 小鼠中的时钟基因。 在模型和限时喂养小鼠中，我们也观察到类似的核心时钟基因独立转录组 这些观察结果为 K01 应用奠定了基础，并引导我们采取非- 核心时钟基因在昼夜节律增强子活性和基因表达的调节中发挥着重要作用 各种正常生理学以及代谢疾病的病理生理学 该提案的目标是。 建立昼夜节律表观基因组重塑与环境挑战之间的联系，并确定和 表征可应用于时间药理学和时间营养的非核心时钟昼夜节律调节器 为了实现这一目标，我计划利用无偏见的全基因组策略。 转录和生物信息学方法绘制增强子景观并识别调控转录 来自成人肝细胞特异性 REV-ERB 的非肝细胞昼夜节律重塑因子 (TF)（目标 1）和 也存在于随意喂养和限时喂养小鼠的肝脏中（目标 2）。实际上，我们的全基因组是公正的。 广泛的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， 此外，我们的无偏转录组分析显示，雌激素相关受体γ（ERRγ）。 ERRγ 的表达在 129S1/SvImJ (129) 小鼠的肝脏中显着较高。值得注意的是，129 只小鼠的肝脏表达水平较高。 B6 小鼠体内产生 DIO 的饮食能够抵抗体重增加和代谢功能障碍。 概念验证研究，我们将确定肝 ERRγ 在对 DIO 的菌株特异性反应中的作用，并扩展 我们针对特定菌株的昼夜节律研究（目标 3）。 在Mitch Lazar博士的指导下，我加强了对转录调控的训练 在这个K01奖期间，我得到了Lazar博士的主要指导和我的指导。 顾问委员会，我将把代谢转录调控的培训扩展到细胞-细胞串扰水平 （单核序列）并获得有关昼夜节律生理学中非核心时钟基因表征的经验 这种培训将使我能够系统地、定量地和功能性地研究分子。 我在科学和职业发展方面的进展将受到监督。 我的咨询委员会负责从博士后实习生过渡到独立研究者。