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

这项提案的总体目标是利用果蝇模型系统来确定 逆转座子元件(RTE)在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 以及阿尔茨海默病(AD)等疾病的进展。我们假设RTE活动的增加 随着年龄的增长，导致体细胞内细胞和组织的动态平衡丧失，从而促进衰老。这又反过来 与AD相关的染色质和RTE活性变化相互作用，加速神经退化。我们将使用 强大的分子和遗传工具，以及果蝇短暂的寿命，来决定如何衰老 和人类AD相关蛋白影响大脑中RTES的活性和染色质状态，并识别新的和 抑制RTE活性和AD相关表型以延长健康寿命和延迟的新基因 苍蝇阿尔茨海默病的发生和发展我们将研究衰老如何影响特定子集中的RTE活动 神经元和胶质细胞，以及这如何影响苍蝇AD的正常衰老过程和神经变性 模特们。利用果蝇模型，我们将使用突变筛选来快速识别新的基因和 控制RTE活性的生理途径，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内进行表型分析，这在人类身上是不可能的， 而且在任何大规模的哺乳动物模型中进行研究都将是昂贵和耗时的。 这项提议的目的是检验衰老和AD相互作用的假设：(I)增加RTE活性和 (Ii)改变成虫大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 在果蝇中分离新的RTE活性抑制因子，以延长健康寿命并推迟发病和死亡 苍蝇AD模型中神经退行性变的研究进展。 为此，我们将使用新的方法在目标1中检查年龄和AD如何改变RTE表达和 用scRNA-seq和RTE动员成虫脑内特定亚群的神经元和神经胶质细胞 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 与具有sc-atac-seq和Cut&Run的RTE相关的状态和表观基因组(与核心B和项目合作 4)，并在Aim 3中使用一种新的正向遗传筛选来识别新的基因和生理途径， 抑制成年果蝇脑内RTE活性，延长寿命，延缓发病 飞来飞去。在与项目1、3和4以及核心C合作中，这些动物和人类的同源物 抑制苍蝇阿尔茨海默病的基因和生理途径将被测试它们是否有能力延迟发病和 小鼠阿尔茨海默病模型神经退行性变及人神经元AD相关细胞表型的研究进展 以及组织培养中的神经胶质细胞。

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.