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昼夜节律重编程的顺式调节逻辑

基本信息

批准号：
10387747
负责人：
India H Reiss
金额：
$ 3.27万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
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/10387747
关键词：
APP-PS1 ARNTL gene Affect Affinity Chromatography Alzheimer&apos s Disease Alzheimer&apos s disease model Alzheimer&apos s disease patient 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 model E-Box Elements Environment Event Exhibits Gene Expression Gene Expression Profile Genes Genetic Transcription Genomics Goals Hour Impaired cognition In Vitro Institutionalization Libraries Location 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 cell type circadian circadian pacemaker experimental study genomic locus in vivo insight mouse model neuron loss novel prevent promoter transcription factor transcriptome sequencing

项目摘要

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.

项目概要阿尔茨海默病 (AD) 是痴呆症最常见的病因，通常伴有昼夜节律目前尚不清楚昼夜节律功能与神经退行性疾病有何关系，但核心昼夜节律的缺失。星形胶质细胞中的转录因子 (TF) BMAL1 会导致 AD 样细胞表型和神经元增加在 AD 小鼠模型的星形胶质细胞中，我们观察到了一种已知的现象。作为昼夜节律重编程，其中一些基因失去昼夜节律表达，而其他基因获得昼夜节律表达该提案的目的是确定星形胶质细胞中的昼夜节律重编程是如何调节的。我在 AD 模型星形胶质细胞中观察到 BMAL1 与疾病特异性 TF 相互作用。为了与新的基因组位置结合并导致新的基因变得有节奏，我将 1) 确定 AD 小鼠模型星形胶质细胞中的 BMAL1 是否发生变化以与重编程基因结合（目的 1) 和 2) 鉴定足以产生昼夜节律基因表达和重编程的 TF 基序组合为了表征目标 1 中的 BMAL1 结合，我有初步数据。 FLEXCC 是一种在细胞类型特定水平上记录 TF 结合的技术，可用于记录 BMAL1 在这个项目中，我将使用 FLEXCC 记录野生型小鼠和星形胶质细胞中 BMAL1 的结合。在目标 2 中，我将识别驱动 AD 中昼夜节律表达的 TF 基序组合。使用大规模并行报告基因检测 (MPRA) 建立小鼠模型并捕获星形胶质细胞转录本该提案将开始确定如何翻译核糖体亲和纯化和RNA测序（TRAP-seq）。 AD 样星形胶质细胞的细胞环境通过建立一种机制来产生昼夜节律重编程。 AD小鼠模型中星形胶质细胞的昼夜节律重编程，我将帮助阐明昼夜节律如何被破坏 AD 的进展并发现可能改变疾病进程的新途径。