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 在衰老和 疾病，并开发抑制RTE活性以延长健康寿命并延迟发作的干预措施 和诸如阿尔茨海默氏病（AD）等疾病的进展。我们假设RTE的活动增加 随着年龄的增长，会导致促进衰老的体细胞中细胞和生物体稳态的丧失。反过来 与AD相关的染色质和RTE活性的变化相互作用，以加速神经退行性。我们将使用 强大的分子和遗传工具以及果蝇的短寿命，以确定衰老 与人类广告相关的蛋白质影响RTE和染色质状态在大脑中的活性，并识别新的和 抑制RTE活性和与广告相关的表型的新型基因，以延长健康的寿命和延迟 苍蝇AD的发作和进展。我们将研究衰老如何影响特定子集中RTE的活性 神经元和神经胶质，以及这如何影响正常的老化过程和苍蝇AD的神经变性 型号。使用果蝇模型，我们将采用诱变筛选来快速识别新基因和 控制RTE活性并使用这些遗传干预措施进行详细的生理途径 表型分析在整个生物体的整个生命周期中，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都会非常昂贵且耗时。 该提案的目的是检验衰老和AD与（i）增加RTE活性和 （ii）在成年苍蝇脑中选择性细胞中改变染色质和表观基因组； （iii）使用正向遗传筛选 在果蝇中，隔离了RTE活性的新抑制剂，这些抑制剂延长了健康的寿命并延迟发作和 飞行AD模型中神经退行性的进展。 为此，我们将使用新方法在AIM 1中检查年龄和广告如何改变RTE表达和 在成年蝇脑中的特定神经元和神经元的特定子集中动员，并动员了 记者（与Core B和Project 4合作），在AIM 2中确定年龄和广告如何改变染色质 与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），在AIM 3中使用新型的前遗传筛选来识别新的基因和生理途径 抑制成人蝇脑的RTE活性，导致寿命延长并延迟发作和进展 飞行广告。与项目1、3和4以及核心C合作的哺乳动物和人类同源物 抑制AD的基因和生理途径将通过其延迟发作的能力进行测试 小鼠AD模型中神经退行性的进展和人类神经元中与AD相关的细胞表型的进展 和组织培养中的神经胶质。