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） 内源性昼夜节律钟，它使细胞生理学与地球的光/暗周期相联系， 生理学的所有方面-从分子到细胞到行为。昼夜节律系统具有不可或缺的作用， 促进细胞在面对环境或内部扰动时的恢复力。昼夜节律紊乱 与心血管疾病、肥胖、肺病和神经精神疾病有关， 细胞时钟机制的基础知识对临床医学的重要性。而 生物钟计时的分子基础已经被精确地定义，生物钟如何对生物钟的变化做出反应。 维持细胞内稳态的环境仍然是未知的。 我们将测试核心生物钟蛋白CLOCK是压力传感器的假设， 通过其内在无序区域中的结构元素将环境信息传递给 昼夜节律计时机制和应激反应机制。我们发现RNA 分子伴侣DDX 3X严格调节CLOCK对环境刺激（如温度）的敏感性 并且对昼夜节律计时和细胞恢复力都有很强的影响。我们将决定如何时钟 机械地与细胞应激机制整合，包括热休克反应。超出其 作为压力传感器的角色，我们将研究昼夜节律系统和时钟如何自适应地组织 蛋白质组的生物化学状态，塑造生物途径的昼夜节律组装， 体内平衡我们将在动物中测试这些细胞数据，通过测量遗传和行为模型， 昼夜节律紊乱和睡眠剥夺重新编程蛋白质组装的节律性。最后我们将 将这些发现应用于人类外周血样本，我们首次发现， 蛋白质合成的节奏。这一建议通过其多尺度和跨学科的方法得到加强 利用PI在时间生物学，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），以及成像和分析开发（Barrett）。 我们的假设是，昼夜节律恢复的分子机制及其在压力下的失效 是一个适应不良的反馈回路的基础， 未对准。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， 对睡眠和昼夜节律紊乱的研究兴趣。我们预计，这一假设的成功测试 将为减轻昼夜节律之间的二价联系提供新的和可操作的目标， 失调以及与之相关的许多人类疾病。 ! !