Project 2: Role of Retrotransposable Element Activity in Drosophila Models of Alzheimer's Disease

项目2：逆转录转座元件活性在阿尔茨海默病果蝇模型中的作用

• 批准号: 10333662
• 负责人: STEPHEN L HELFAND
• 金额: $ 57.97万
• 依托单位: BROWN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2026-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10333662
• 关键词: ATAC-seq; Adult; Affect; Age; Aging; Alzheimer' s Disease; Alzheimer' s disease model; Alzheimer' s disease risk; Bioinformatics; Biological Models; Brain; Cells; Chromatin; Collaborations; Consumption; Development; Disease; Disease Progression; Drosophila genus; Elements; Functional disorder; Genes; Genetic Screening; Genetic Transcription; Goals; Heterochromatin; Homeostasis; Human; Inflammation; Intervention; Life Extension; Longevity; Maintenance; Mammals; Methods; Modeling; Molecular Genetics; Mus; Mutagenesis; Nerve Degeneration; Nervous system structure; Neuroglia; Neurons; Organism; Pathway interactions; Phenotype; Physiological; Process; Proteins; Regulation; Reporter; Retrotransposon; Role; Series; Somatic Cell; Testing; Time; Tissues; Work; age effect; age related; brain cell; cell type; epigenome; fly; gene therapy; innovation; normal aging; novel; single cell analysis; single-cell RNA sequencing; tissue culture; tool

The overall goals of this proposal are to exploit the Drosophila model system to determine the role of retrotransposable elements (RTEs) in the progression of cellular dysfunction that occurs during aging and disease, and to develop interventions that suppress RTE activity to extend healthy life span and delay the onset and progression of diseases such as Alzheimer's Disease (AD). We hypothesize that increased activity of RTEs with age leads to loss of cellular and organismal homeostasis in somatic cells promoting aging. This in turn interacts with AD-related changes in chromatin and RTE activity to accelerate neurodegeneration. We will use the powerful molecular and genetic tools, as well as the short life span of Drosophila, to determine how aging and human AD-related proteins affect the activity of RTEs and chromatin state in the brain and identify new and novel genes that repress RTE activity and AD-related phenotypes in order to extend healthy life span and delay the onset and progression of fly AD. We will examine how aging affects the activity of RTEs in specific subsets of neurons and glia, and in turn, how this affects both normal aging processes and neurodegeneration in fly AD models. Using the Drosophila model, we will employ mutagenesis screens to rapidly identify new genes and physiological pathways controlling RTE activity and use these genetic interventions to perform detailed phenotypic analyses over the entire life span of the organism, something that would not be possible in humans, and would be prohibitively expensive and time-consuming to do in mammalian models on any large scale. The aims of this proposal are to test the hypotheses that aging and AD interact to (i) increase RTE activity and (ii) alter chromatin and the epigenome in selective cells in the adult fly brain; and (iii) use forward genetic screens in Drosophila to isolate new suppressors of RTE activity that extend healthy life span and delay the onset and progression of neurodegeneration in fly AD models. To do this we will use new methods to examine in Aim 1 how age and AD change RTE expression and mobilization in specific subsets of neurons and glia in the adult fly brain with scRNA-seq and RTE mobilization reporters (in collaboration with Core B and Project 4), in Aim 2 determine how age and AD change the chromatin state and epigenome related to RTEs with sc-ATAC-seq and CUT&RUN (in collaboration with Core B and Project 4), and in Aim 3 use a novel forward genetic screen to identify new genes and physiological pathways that suppress RTE activity in adult fly brains, resulting in extension of life span and delaying the onset and progression of fly AD. In collaboration with Projects 1, 3, and 4 and Core C the mammalian and human homologs of these genes and physiological pathways that suppress fly AD will be tested for their ability to delay the onset and progression of neurodegeneration in mouse AD models and AD-related cellular phenotypes in human neurons and glia in tissue culture.

该提案的总体目标是利用果蝇模型系统来确定 逆转录转座元件（RTE）在衰老和衰老过程中发生的细胞功能障碍进展中的作用 疾病，并制定抑制 RTE 活动的干预措施，以延长健康寿命并延迟发病 以及阿尔茨海默病 (AD) 等疾病的进展。我们假设 RTE 的活性增加 随着年龄的增长，体细胞中的细胞和有机体稳态丧失，从而促进衰老。这反过来 与 AD 相关的染色质和 RTE 活性变化相互作用，加速神经退行性变。我们将使用 强大的分子和遗传工具，以及果蝇的短暂寿命，可以确定衰老的方式 和人类 AD 相关蛋白影响大脑中 RTE 的活性和染色质状态，并识别新的和 抑制 RTE 活性和 AD 相关表型的新基因，以延长健康寿命并延迟 果蝇 AD 的发生和进展。我们将研究衰老如何影响特定亚群中 RTE 的活性 神经元和神经胶质细胞的变化，以及这如何影响果蝇 AD 的正常衰老过程和神经退行性变 模型。使用果蝇模型，我们将采用诱变筛选来快速识别新基因并 控制 RTE 活动的生理途径，并使用这些遗传干预来执行详细的 对生物体整个生命周期进行表型分析，这在人类中是不可能的， 在任何大规模的哺乳动物模型中进行这项工作都将非常昂贵且耗时。 该提案的目的是测试衰老和 AD 相互作用的假设：(i) 增加 RTE 活动和 (ii) 改变成年果蝇大脑中选择性细胞的染色质和表观基因组； (iii) 使用正向遗传筛选 在果蝇中分离出 RTE 活性的新抑制剂，可延长健康寿命并延缓发病 果蝇 AD 模型中神经退行性变的进展。 为此，我们将在目标 1 中使用新方法来检查年龄和 AD 如何改变 RTE 表达和 通过 scRNA-seq 和 RTE 动员对成年果蝇大脑中特定神经元和神经胶质亚群进行动员 记者（与核心 B 和项目 4 合作），在目标 2 中确定年龄和 AD 如何改变染色质 使用 sc-ATAC-seq 和 CUT&RUN 与 RTE 相关的状态和表观基因组（与 Core B 和 Project 合作） 4），并在目标 3 中使用新颖的正向遗传筛选来识别新的基因和生理途径， 抑制成年果蝇大脑中的 RTE 活动，从而延长寿命并延缓发病和进展 飞广告。与项目 1、3、4 和 Core C 合作，这些的哺乳动物和人类同源物 抑制果蝇 AD 的基因和生理途径将被测试其延迟发病的能力， 小鼠 AD 模型中神经变性的进展和人类神经元中 AD 相关细胞表型 和组织培养中的神经胶质细胞。