CELL-TYPE SPECIFIC ROLE OF CIRCADIAN-DEPENDENT TRANSCRIPTION IN FENTANYL-INDUCED SYNAPTIC AND BEHAVIORAL PLASTICITY

昼夜节律依赖性转录在芬太尼诱导的突触和行为可塑性中的细胞类型特异性作用

• 批准号: 10370036
• 负责人: Ryan W Logan
• 金额: $ 118.75万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-23 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10370036
• 关键词: CELL; TYPE; SPECIFIC; ROLE; CIRCADIAN

PROJECT SUMMARY Sleep and circadian disruption are highly prevalent in Alzheimer’s disease, emerging decades prior to cognitive decline. Evidence from animals and humans suggests these disruptions directly lead to Alzheimer’s disease pathology that further exacerbate sleep and circadian dysfunctions. An important breakthrough for studying these connections was the recent development of a genetically diverse mouse panel that incorporates high-risk familial Alzheimer’s disease mutations (termed AD-BXD) that recapitulate key aspects of human Alzheimer’s pathophysiology, including aging-related neurodegeneration, progressive cognitive deficits, and sleep disruption. Using these mice, identified a new marker of genetic vulnerability to cognitive decline and sleep disruption in Alzheimer’s disease: the transient-receptor potential nonselective cation channel type 3 (TRPC3). TRPC3 has already been implicated as a target for modifying the development of normal cognitive aging and Alzheimer’s disease. We found that viral-mediated knockdown of TRPC3 diminished amyloid load and enhanced cognition in susceptible AD-BXD mice, providing the preclinical basis for investigating the mechanistic links that connect sleep disruption and cognitive decline in Alzheimer’s disease. However, these methods do not offer the biology by which TRPC3 moderate disease-related pathology. We propose to comprehensively map the spatial location and expression of TRPC3 to identify whether this localization changes in key brain regions related to sleep and cognition due to Alzheimer’s pathology. We developed a new approach to rapidly image multiple cell-types and markers of Alzheimer’s neurodegeneration within a whole brain in three-dimensional (3D) space at single-cell resolution. Our ultra-fast high-resolution confocal ribbon-scanning approach reaches diffraction limited resolution (~200nm) and collects 3D rendered whole brain maps in less than 24-hours. The goals of the supplement are to take advantage of the discovery of TRPC3 as a new target for Alzheimer’s-related changes in cognition and sleep and use our innovative tools to answer fundamental questions: In mice, 1) Is TRPC3 located in disease-related brain areas linked to cognition and sleep?; and 2) Does TRPC3 interact with biological hallmarks of Alzheimer’s disease brain pathology, including amyloid beta and hyperphosphorylated Tau? We will use human postmortem brains from Alzheimer’s disease patients to ask, 3) Is brain TRPC3 expression altered in Alzheimer’s disease? We predict TRPC3 localizes to subregions of the hypothalamus associated with sleep and hippocampal and cortical regions associated with cognition, and this distribution will be differentially impacted by sleep deprivation in susceptible vs. resilient mice. In human brains, we expect that TRPC3 expression will be higher in advanced Alzheimer’s disease patient brains and display altered rhythmicity. Ultimately, using our novel suite of genetic, imaging, and computational tools, we will we will answer longstanding questions about how changes in sleep and cognitive decline are linked to Alzheimer’s disease pathology.

项目概要 睡眠和昼夜节律紊乱在阿尔茨海默病中非常普遍，其出现早于认知疾病数十年。 衰退。来自动物和人类的证据表明这些干扰直接导致阿尔茨海默病 进一步加剧睡眠和昼夜节律功能障碍的病理学。这些研究的重要突破 连接是最近开发的遗传多样性小鼠组，其中包含高风险家族性 阿尔茨海默病突变（称为 AD-BXD）概括了人类阿尔茨海默病的关键方面 病理生理学，包括与衰老相关的神经退行性变、进行性认知缺陷和睡眠中断。 利用这些小鼠，我们发现了一种新的标志物，该标志物表明小鼠易受认知能力下降和睡眠干扰的影响。 阿尔茨海默病：瞬时受体电位非选择性阳离子通道 3 型 (TRPC3)。 TRPC3 有 已经被认为是改变正常认知衰老和阿尔茨海默病发展的目标 疾病。我们发现病毒介导的 TRPC3 敲低可减少淀粉样蛋白负荷并增强认知能力 在易感 AD-BXD 小鼠中，为研究连接的机制链接提供临床前基础 阿尔茨海默病的睡眠中断和认知能力下降。然而，这些方法不提供生物学 TRPC3 通过 TRPC3 调节疾病相关的病理。我们建议全面绘制空间图 TRPC3 的位置和表达，以确定这种定位是否在关键大脑区域发生变化 由于阿尔茨海默病的病理学，与睡眠和认知有关。我们开发了一种快速成像的新方法 全脑三维 (3D) 中的多种细胞类型和阿尔茨海默氏症神经变性标记物 单细胞分辨率的空间。我们的超快高分辨率共焦带状扫描方法达到 衍射极限分辨率（~200nm），并在 24 小时内收集 3D 渲染的全脑图。这 该补充剂的目标是利用 TRPC3 的发现作为阿尔茨海默病相关疾病的新靶点 认知和睡眠的变化，并使用我们的创新工具来回答基本问题：在小鼠中，1) 是 TRPC3 位于与认知和睡眠相关的疾病相关大脑区域？ 2) TRPC3 是否与 阿尔茨海默病脑病理学的生物学特征，包括β淀粉样蛋白和过度磷酸化 头？我们将使用阿尔茨海默病患者的死后大脑来询问，3）大脑是TRPC3吗？ 阿尔茨海默氏病的表达发生改变？我们预测 TRPC3 定位于下丘脑的亚区域 与睡眠相关以及与认知相关的海马和皮质区域，这种分布将 易感小鼠与弹性小鼠受到睡眠剥夺的影响不同。在人类大脑中，我们期望 TRPC3 在晚期阿尔茨海默病患者大脑中的表达会更高，并表现出节律性改变。 最终，使用我们新颖的遗传、成像和计算工具套件，我们将回答长期存在的问题 关于睡眠变化和认知能力下降如何与阿尔茨海默病病理学相关的问题。