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.

这项提议的总体目标是利用果蝇模型系统来确定 逆转录转座因子（RTEs）在衰老过程中发生的细胞功能障碍进展中的作用， 疾病，并开发抑制RTE活动的干预措施，以延长健康寿命并推迟发病 和疾病如阿尔茨海默病（AD）的进展。我们推测，RTE活性的增加 随着年龄的增长，导致体细胞中细胞和有机体稳态的丧失，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，加速神经变性。我们将使用 强大的分子和遗传工具，以及果蝇的短暂寿命，来确定衰老如何 和人AD相关蛋白影响RTEs的活性和脑中的染色质状态，并鉴定新的和 抑制RTE活性和AD相关表型的新基因，以延长健康寿命和延迟 苍蝇AD的发病和进展。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，以及反过来，这如何影响正常的衰老过程和神经退行性疾病的苍蝇 模型使用果蝇模型，我们将采用诱变筛选来快速鉴定新基因， 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 在生物体的整个生命周期中进行表型分析，这在人类中是不可能的， 并且在任何大规模的哺乳动物模型中进行都是极其昂贵和耗时的。 本提案的目的是检验衰老和AD相互作用的假设：（i）增加RTE活性， (ii)改变成年果蝇脑中选择性细胞的染色质和表观基因组;以及（iii）使用正向遗传筛选 在果蝇中分离出新的RTE活性抑制因子，延长健康寿命，延迟发病， 果蝇AD模型中神经变性的进展。 为了做到这一点，我们将使用新的方法来检查目标1中年龄和AD如何改变RTE表达， 用scRNA-seq和RTE动员在成年果蝇脑中的神经元和神经胶质细胞的特定亚群中的动员 报告者（与核心B和项目4合作），在Aim 2中确定年龄和AD如何改变染色质 与使用sc-ATAC-seq和CUT&RUN的RTE相关的状态和表观基因组（与Core B和Project 4），并在目标3中使用一种新的正向遗传筛选来鉴定新的基因和生理途径， 抑制成年苍蝇大脑中的RTE活动，从而延长寿命并延迟发病和进展 飞AD与项目1、3和4以及核心C合作， 将测试抑制苍蝇AD的基因和生理途径延迟发病的能力， 小鼠AD模型中神经变性的进展和人类神经元中AD相关的细胞表型 和神经胶质细胞。