A genomic characterization of the response to sleep loss

睡眠不足反应的基因组特征

• 批准号: 10928421
• 负责人: Robert W Greene
• 金额: $ 41万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-21 至 2024-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10928421
• 关键词: ADORA1 gene; ATAC-seq; Adenosine; Adenosine A1 Receptor; Adenosine Kinase; Affinity; Attenuated; Binding; Binding Sites; Biological; Brain; Cell physiology; Cells; Chromatin; Chronic; DNA; DNA Binding; Dissociation; Enhancers; Foundations; Future; Gene Expression; Genes; Genomics; Growth; Homeostasis; Hour; Knock-out; Mediation; Mediator; Mental disorders; Metabolic syndrome; Mitochondria; Molecular; Mus; Nerve Degeneration; Neurobiology; Neuroglia; Neurons; Non-Essential Amino Acid; Nucleic Acids; Ontology; Pathologic; Pathology; Pathway interactions; Process; Proteins; Purinergic P1 Receptors; Reaction; Receptor Activation; Recovery; Research; Resolution; Ribosomes; Role; Signal Pathway; Signal Transduction; Site; Sleep; Sleep Deprivation; Sleep disturbances; Slow-Wave Sleep; Specificity; Synapses; Testing; Therapeutically Targetable; Time; Tissue-Specific Gene Expression; Tissues; Untranslated RNA; Validation; Work; awake; cell type; cellular targeting; design; differential expression; extracellular; frontal lobe; genome wide association study; insight; mRNA Expression; mutant; myocyte-specific enhancer-binding-factor 2C; neuronal excitability; neurotransmission; permissiveness; promoter; receptor; response; sleep regulation; transcription factor; transcriptome

Sleep gates, or is permissive to, the resolution of sleep need. The resolution of sleep need reflects sleep function induced in response to waking (sleep homeostasis). Much is now known about the control of sleep/wake state expression and duration, but sleep function is less well understood. Significant differential expression of genes (DEG’s; 6000/13000 expressed in bulk tissue from frontal cortex; FC) in response to 6 hours of sleep deprivation (SD) has been observed. Of particular significance for CNS tissue, the sleep modulated gene ontology supports sleep control of neuronal excitability and transmission. Sleep dependent modulation of cellular function is likely to vary depending on different cell-type specific roles yet, little is understood about the cell-type specificity of sleep DEGs and their implications for biological intra- and inter-cellular pathways responsible for the SD response. Ado may be implicated as an inter- cellular (glia to neuron) mediator of sleep need since activation of its receptor, ADORA1 is required for sleep-homeostatic slow wave activity (SWA) but its role in sleep-related gene expression has not been examined. The following specific aims are proposed to characterize the genomic responses to sleep loss with respect to the cell-type specificity of DEGs and the pathways determining their SD response at an intercellular and intracellular level, in the mouse FC. 1.The differential expression of cell-type specific mRNA expression in response to sleep-loss will be compared to ad-libitum sleep condition and to recovery sleep condition to characterize the cell-type, specific and shared gene expression responses to sleep loss and recovery in FC tissue of C57 BL/6J mice. 2. Identification of cell-type specific open chromatin induced by sleep deprivation and sleep loss recovery will be made using ATACseq to identify potential enhancer and promoter sites important to the sleep loss response. 3. The role of Ado activation of ADORA1 receptors in the cell-type specific genomic response to sleep loss will be examined using a conditional Adora1 knockout that is unresponsive to Ado and a glial conditional Adk knockout with chronically elevated brain Ado and elevated SWA during waking and sleep. The first aim may provide fundamental insight into the cellular targets of sleep function with respect to cell-type specificity and differential genomic ontology in response to sleep loss. The second aim is designed to identify DNA loci as potential, cell type-specific, enhancer and/or promoter sites. Identification of these sites is important for characterization of genomic pathways activated by SD and further, is a first step towards understanding and prioritizing GWAS identified loci in non-coding DNA regions for future validation. The third aim will test Ado’s role as a mediator of the genomic changes observed in response to sleep loss and whether or not its mediation of homeostatic sleep SWA can be dissociated from the sleep loss transcriptome.

睡眠控制或允许睡眠需求的解决。 关于睡眠/觉醒状态的控制现在已经了解很多。 表达和持续时间，但睡眠功能尚不清楚。 基因的显着差异表达（DEG；6000/13000 在额叶皮层的大块组织中表达；FC） 据观察，6 小时睡眠剥夺 (SD) 对中枢神经系统组织具有特别重要的意义。 睡眠调节基因本体支持神经兴奋性和传导的睡眠控制。 睡眠依赖性细胞功能调节可能会根据不同细胞类型的特定作用而变化 然而，人们对睡眠 DEG 的细胞类型特异性及其对生物体内和体内的影响知之甚少。 负责 SD 反应的细胞间通路。 Ado 可能被认为是睡眠需求的细胞间（神经胶质细胞到神经元）介体，因为其受体被激活， ADORA1 是睡眠稳态慢波活动 (SWA) 所必需的，但它在睡眠相关基因表达中的作用 尚未经过检查。 提出以下具体目标来描述基因组对睡眠不足的反应 DEG 的细胞类型特异性以及决定其在细胞间和细胞间的 SD 反应的途径 细胞内水平，在小鼠 FC 中。 1.比较细胞类型特异性mRNA表达对睡眠缺失的差异表达 随意睡眠条件和恢复睡眠条件来表征细胞类型、特定和共享 C57 BL/6J 小鼠 FC 组织中睡眠缺失和恢复的基因表达反应。 2. 睡眠剥夺诱导的细胞类型特异性开放染色质和睡眠缺失恢复的鉴定 使用 ATACseq 来识别对睡眠不足反应重要的潜在增强子和启动子位点。 3. Ado 激活 ADORA1 受体在细胞类型特异性基因组对睡眠不足反应中的作用 使用对 Ado 无反应的条件 Adora1 敲除和神经胶质条件 Adk 进行检查 在清醒和睡眠期间，大脑 Ado 和 SWA 长期升高，从而导致敲除。 第一个目标可能提供对睡眠功能相对于细胞类型的细胞目标的基本了解 第二个目标是识别针对睡眠不足的特异性和差异基因组本体。 DNA 位点作为潜在的、细胞类型特异性的、增强子和/或启动子位点，这些位点的鉴定。 对于 SD 激活的基因组通路的表征非常重要，并且是迈向的第一步 理解并优先考虑 GWAS 确定的非编码 DNA 区域中的位点以供将来验证。 目标将测试 Ado 作为因睡眠不足而观察到的基因组变化的中介者的作用，以及是否 或不它对稳态睡眠的介导 SWA 可以与睡眠缺失转录组分离。