Deciphering the cis-regulatory logic of circadian reprogramming in a mouse model of Alzheimer's Disease

破译阿尔茨海默病小鼠模型中昼夜节律重编程的顺式调节逻辑

• 批准号: 10598018
• 负责人: India H Reiss
• 金额: $ 3.36万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10598018
• 关键词: APP-PS1; ARNTL gene; Affect; Affinity Chromatography; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease patient; Arrhythmia; Astrocytes; Automobile Driving; Bar Codes; Binding; Binding Sites; Biological Assay; Brain; Cells; ChIP-seq; Circadian Dysregulation; Circadian Rhythms; Circadian gene expression; Clock protein; DNA Binding; Data; Dementia; Disease; Disease Progression; Disease model; E-Box Elements; Environment; Event; Exhibits; Gene Expression; Gene Expression Profile; Genes; Genetic Transcription; Genomics; Goals; Hour; Impaired cognition; In Vitro; Institutionalization; Libraries; Logic; Modeling; Molecular; Mus; Nerve Degeneration; Neurons; Nuclear; Nucleic Acid Regulatory Sequences; Oxidative Stress; Pathway interactions; Patients; Pattern; Periodicity; Phenotype; Protocols documentation; Reporter; Reporter Genes; Ribosomes; Signal Transduction; Sleep disturbances; Technology; Testing; Transcript; Translating; Wild Type Mouse; astrogliosis; candidate identification; cell type; circadian; circadian pacemaker; experimental study; genomic locus; in vivo; insight; mouse model; neuron loss; novel; prevent; promoter; transcription factor; transcriptome sequencing

PROJECT SUMMARY Alzheimer’s Disease (AD), the most common origin of dementia, is frequently accompanied by circadian rhythm disruption. It is not clear how circadian function relates to neurodegeneration, but deletion of the core circadian transcription factor (TF) BMAL1 in astrocytes causes both an AD-like cellular phenotype and increased neuron loss after oxidative stress. Within astrocytes of a mouse model of AD, we have observed a phenomenon known as circadian reprogramming, in which some genes lose circadian expression while other genes gain circadian expression. The goal of this proposal is to determine how circadian reprogramming is regulated in astrocytes of our AD mouse model. I hypothesize that within AD-model astrocytes, BMAL1 interacts with disease-specific TFs to bind to new genomic locations and cause new genes to become rhythmic. To test this hypothesis, I will 1) determine whether BMAL1 changes in astrocytes of our AD mouse model to bind to reprogrammed genes (Aim 1), and 2) identify TF motif combinations sufficient to produce circadian gene expression and reprogramming in astrocytes of our AD mouse model (Aim 2). To characterize BMAL1 binding in Aim 1, I have preliminary data that FLEXCCs, a technology to record TF binding on a cell type specific level, can be used to record BMAL1 binding. For this project, I will use FLEXCCs to record BMAL1 binding in astrocytes of both wildtype mice and in our AD mouse model. In Aim 2, I will identify TF motif combinations that drive circadian expression in our AD mouse model using a Massively Parallel Reporter Assay (MPRA) and capture astrocytic transcripts with translating ribosome affinity purification and RNA-sequencing (TRAP-seq). This proposal will begin to determine how the cellular environment of AD-like astrocytes generates circadian reprogramming. By establishing a mechanism of circadian reprogramming in astrocytes in a mouse model of AD, I will help elucidate how circadian disruption progresses in AD and uncover new pathways to potentially alter disease course.

项目摘要 阿尔茨海默病（Alzheimer's Disease，AD）是痴呆症最常见的病因，常伴有昼夜节律 破坏目前尚不清楚昼夜节律功能如何与神经退行性变相关，但核心昼夜节律的缺失 星形胶质细胞中的转录因子（TF）BMAL 1导致AD样细胞表型和增加的神经元细胞数量。 氧化应激后的损失。在AD小鼠模型的星形胶质细胞内，我们观察到一种已知的现象， 如昼夜节律重编程，其中一些基因失去昼夜节律表达，而另一些基因获得昼夜节律表达。 表情这项计划的目标是确定昼夜节律重编程是如何在星形胶质细胞中调节的， 我们的AD小鼠模型我假设在AD模型星形胶质细胞中，BMAL 1与疾病特异性TF相互作用 与新的基因组位置结合，使新的基因变得有节奏。为了验证这个假设，我将1） 确定BMAL 1是否在我们的AD小鼠模型的星形胶质细胞中改变以结合重编程基因（Aim 1），和2）鉴定足以产生昼夜节律基因表达和重编程的TF基序组合， 我们的AD小鼠模型的星形胶质细胞（Aim 2）。为了表征Aim 1中BMAL 1的结合，我有初步数据 FLEXCC是一种在细胞类型特异性水平上记录TF结合的技术，可用于记录BMAL 1 约束力在这个项目中，我将使用FLEXCC记录野生型小鼠和小鼠星形胶质细胞中BMAL 1的结合。 我们的AD小鼠模型在目标2中，我将确定驱动AD昼夜节律表达的TF基序组合 使用大规模平行报告基因测定（MPRA）的小鼠模型，并使用 翻译核糖体亲和纯化和RNA测序（TRAP-seq）。本提案将开始确定如何 AD样星形胶质细胞的细胞环境产生昼夜节律重编程。通过建立一种机制， 在AD小鼠模型的星形胶质细胞中的昼夜节律重编程，我将帮助阐明昼夜节律的破坏是如何 并发现可能改变疾病进程的新途径。