Investigating Circadian Mechanisms of Cellular Resilience: Rhythmic Condensates, Disorder, and Stress

研究细胞弹性的昼夜节律机制：节律凝聚、紊乱和压力

• 批准号: 10614584
• 负责人: Jonathan Oren Lipton
• 金额: $ 41.88万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-05-01 至 2025-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10614584
• 关键词: ARNTL gene; Affect; Aging; Amino Acids; Animals; Articulation; Behavioral; Behavioral Model; Biochemical; Biochemistry; Biological; Biology; Blood specimen; Brain; Cardiovascular Diseases; Cardiovascular system; Cell Line; Cell physiology; Cells; Cellular Stress; Cellular biology; Childhood; Chronobiology; Circadian Dysregulation; Circadian Rhythms; Circadian desynchrony; Clinical; Clinical Medicine; Clock protein; Collaborations; Compensation; Coronary Arteriosclerosis; Darkness; Data; Data Set; Diabetes Mellitus; Disease; Disease Pathway; Disease susceptibility; Elements; Engineering; Environment; Failure; Feedback; Gene Expression; Genes; Genetic Models; Genetic Transcription; Genomics; Goals; Heat-Shock Response; Homeostasis; Human; Image; Knowledge; Laboratories; Life; Light; Link; Liver; Lung diseases; Maintenance; Maps; Measures; Mediating; Medicine; Metabolic; Modeling; Modernization; Molecular; Molecular Chaperones; Movement; Mus; Neurologist; Obesity; Output; Pathway interactions; Patients; Periodicity; Phase; Physical condensation; Physicians; Physiology; Planet Earth; Post-Translational Protein Processing; Predisposition; Protein Biosynthesis; Proteins; Proteome; Proteomics; RNA; Reporter; Research; Role; Secondary to; Sleep; Sleep Deprivation; Sleep Disorders; Sleep disturbances; Solubility; Starvation; Stimulus; Stress; System; Temperature; Testing; Therapeutic; Time; Training; Translating; Translations; assay development; biological adaptation to stress; biophysical properties; cellular resilience; circadian; circadian biology; circadian pacemaker; experience; experimental study; human disease; individual variation; inhibitor; interdisciplinary approach; interest; knock-down; molecular clock; multicatalytic endopeptidase complex; multidisciplinary; nervous system disorder; neuropsychiatric disorder; neuropsychiatry; novel; peripheral blood; pharmacologic; phosphoproteomics; programs; promote resilience; resilience; response; sensor; stressor; structural determinants; superresolution imaging; transcription factor; transmission process

PROJECT SUMMARY (ABSTRACT) The endogenous circadian clock that synchronizes cellular physiology with the earth's light/dark cycle affects all aspects of physiology – from molecular to cellular to behavioral. The circadian system has an integral role in promoting cellular resilience in the face of environmental or internal perturbation. Circadian disruption has been linked with cardiovascular disease, obesity, pulmonary disease, and neuropsychiatric disorders, highlighting the importance of fundamental knowledge of cellular clock mechanisms for clinical medicine. While the molecular basis for circadian timekeeping has been exquisitely defined, how the clock responds to the environment to maintain cellular homeostasis remains mostly unknown. We will test the hypothesis that the core circadian clock protein CLOCK is a stress sensor that transmits environmental information through structural elements in its intrinsically disordered region to both the circadian timekeeping mechanism and the stress-response machinery. We have discovered that the RNA chaperone DDX3X critically regulates the sensitivity of CLOCK to environmental stimuli such as temperature and has potent effects on both circadian timekeeping and cellular resilience. We will determine how CLOCK mechanistically integrates with the cellular stress machinery, including the heat shock response. Beyond its role as a stress sensor, we will investigate how the circadian system and CLOCK adaptively organize the biochemical state of the proteome, sculpting the circadian assembly of biological pathways involved in homeostasis. We will test these cellular data in animals by measuring how genetic and behavioral models of circadian disruption and sleep deprivation re-program the rhythmicity of protein assemblies. Finally, we will apply these discoveries to human peripheral blood samples in which we have, for the first time, identified rhythms of protein synthesis. This proposal is strengthened by its multi-scaled and interdisciplinary approaches that harness the expertise of the PI in chronobiology, cell biology, and biochemistry in collaboration with experts in human circadian biology (Klerman), proteomics and phosphoproteomics (Asara), live cell and super-resolution imaging (Chen), and imaging and assay development (Barrett). Our hypothesis is that the molecular mechanisms for circadian resilience and their failure during stress underlie a maladaptive feedback loop that connects integrated cellular stress responses with circadian misalignment. Determining the mechanisms by which the circadian clock supervises, senses, and responds to environmental stimuli is a crucial challenge for linking circadian disruption to disease mechanisms. My clinical background and experience as a pediatric neurologist trained in sleep medicine supports and informs my research interest in sleep and circadian disorders. We anticipate that the successful testing of this hypothesis will offer novel and pharmacologically actionable targets for mitigating the bivalent link between circadian misalignment and the many human disorders associated with it. ! !

项目概要（摘要） 使细胞生理学与地球光/暗周期同步的内源性生物钟影响 生理学的各个方面——从分子到细胞再到行为。昼夜节律系统在以下方面发挥着不可或缺的作用： 促进细胞面对环境或内部扰动时的恢复能力。昼夜节律被扰乱 与心血管疾病、肥胖、肺部疾病和神经精神疾病有关，强调 细胞时钟机制基础知识对临床医学的重要性。虽然 昼夜节律计时的分子基础已经被精确定义，时钟如何响应 维持细胞稳态的环境仍然大多未知。 我们将检验以下假设：核心生物钟蛋白 CLOCK 是一种压力传感器， 通过其本质上无序区域中的结构元素将环境信息传输到 昼夜节律计时机制和压力反应机制。我们发现RNA 伴侣 DDX3X 严格调节 CLOCK 对温度等环境刺激的敏感性 并对昼夜节律计时和细胞弹性有强大的影响。我们将确定如何CLOCK 与细胞应激机制机械地整合，包括热休克反应。超越其 作为压力传感器，我们将研究昼夜节律系统和时钟如何自适应地组织 蛋白质组的生化状态，塑造参与的生物途径的昼夜节律组装 体内平衡。我们将通过测量遗传和行为模型如何在动物身上测试这些细胞数据 昼夜节律紊乱和睡眠不足会重新编程蛋白质组装的节律。最后，我们将 将这些发现应用于人类外周血样本，我们首次在其中鉴定出 蛋白质合成的节律。该提案通过其多尺度和跨学科方法得到加强 与 PI 合作，利用时间生物学、细胞生物学和生物化学方面的专业知识 人类昼夜节律生物学（Klerman）、蛋白质组学和磷酸蛋白质组学（Asara）、活细胞和 超分辨率成像（Chen）以及成像和检测开发（Barrett）。 我们的假设是昼夜节律恢复力及其在压力下失败的分子机制 是一个将细胞应激反应与昼夜节律联系起来的适应不良反馈循环的基础 错位。确定生物钟监督、感知和响应的机制 环境刺激是将昼夜节律破坏与疾病机制联系起来的一个关键挑战。我的临床 作为一名接受过睡眠医学培训的儿科神经科医生的背景和经验支持并告知我 对睡眠和昼夜节律紊乱的研究兴趣。我们预计这一假设的成功检验 将提供新颖且药理学上可行的目标，以减轻昼夜节律之间的二价联系 失调以及与之相关的许多人类疾病。 ！ ！