The role of Sirtuins in neurodegenerative disease

这项建议试图描述阿尔茨海默病(AD)模型的转录组和表观基因组 黑腹果蝇。该项目还建议调查sirtuins在发展和发展中的作用。 AD的进展，并了解其已报道的神经保护作用的分子机制。我们将使用 结合基于下一代测序技术的计算生物学方法来识别两者 在AD期间发生的转录和染色质状态的变化，并决定这些变化是如何被 Sirtuin蛋白的过度表达或敲除。 这个项目的具体目标是探索AD以异染色质丢失为特征的假设， 增加基因组的不稳定性和异常转录，而sirtuin的过度表达可以稳定这些过程 并延缓疾病的发展。这一假说是基于最近几项将sirtuin生物学的各个方面与 神经退行性疾病。目标1将确定调节sirtuin表达如何影响转录图谱 Fly AD模型。我们将使用rna-seq方法来研究基因表达、小rna种类和转座。 元素在AD模型中的表达，并观察SIRT6和SIRT6基因过表达或敲除对这些元素的影响 SIRT1。目的2将探讨调节sirtuin表达如何影响苍蝇AD模型的染色质景观。 具体地说，我们将使用atac-seq来确定染色质的可及性，并确定 已知的sirtuins靶向的组蛋白标记，包括H3K9ac、H3K56ac和H4K16ac。我们还将 确定过度表达或敲除sirtuins如何影响这些AD染色质图谱。 这些实验将产生丰富而全面的数据集，对这些数据集的分析将产生对这两个问题的见解 Sirtuin生物学与阿尔茨海默病的病因和进展。我们还希望利用 果蝇模型系统，包括成本、时间和基因表达的精确控制，以验证计算 在活体内进行生物学观察，并与传统的遗传学实验进行随访。

This proposal seeks to characterize the transcriptome and epigenome of models of Alzheimer’s disease (AD) in Drosophila melanogaster. The project further proposes to investigate the role of sirtuins in the development and progression of AD, and to understand the molecular mechanism of their reported neuroprotective effects. We will use a computational biology approach with a combination of next-generation sequencing-based techniques to identify both transcriptional and chromatin state changes that take place during AD and determine how these are modified by overexpression or knockout of sirtuin proteins. The specific aims of this project seek to explore the hypothesis that AD is characterized by a loss of heterochromatin, increased genomic instability, and aberrant transcription, and that sirtuin overexpression can stabilize these processes and delay disease progression. This hypothesis is based on several recent studies linking aspects of sirtuin biology to neurodegenerative disease. Aim 1 will determine how modulating sirtuin expression affects the transcriptional profile of fly AD models. We will use an RNA-seq approach to study gene expression, small RNA species, and transposable element expression in AD models, and observe how these are affected upon overexpression or knockout of Sirt6 and Sirt1. Aim 2 will explore how modulating sirtuin expression affects the chromatin landscape of fly AD models. Specifically, we will determine chromatin accessibility using ATAC-seq, and determine the pattern and abundance of histone marks known to be targeted by sirtuins, including H3K9ac, H3K56ac, and H4K16ac by ChIP-seq. We will also determine how these AD chromatin profiles are affected by overexpressing or knocking out sirtuins. These experiments will generate rich and comprehensive data sets, analysis of which will yield insights into both sirtuin biology and Alzheimer’s disease etiology and progression. We also expect to leverage the strengths of the Drosophila model system, including cost, time, and precise control of gene expression, to validate computational biology observations in vivo and follow up with traditional genetic experiments.