A study of ZCCHC17 regulation of synaptic genes in Alzheimers disease

ZCCHC17对阿尔茨海默病突触基因调控的研究

• 批准号: 8757603
• 负责人: Andrew Franklin Teich
• 金额: $ 12万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-09-01 至 2016-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8757603
• 关键词: Adult; Affect; Alzheimer' s Disease; Amyloid; Amyloid Proteins; Amyloid beta-Protein Precursor; Atrophic; Behavior; Binding; Bioinformatics; Brain; Cell Death; Cell Nucleus; Cells; ChIP-seq; Computer Analysis; DNA; Data; Dementia; Disease; Down-Regulation; Environment; Functional disorder; Future; Gene Expression; Gene Expression Regulation; Genes; Genetic Transcription; Goals; Health; Heart; Hippocampus (Brain); Human; Lasers; Lead; Link; Memory; Memory Loss; Messenger RNA; Molecular Profiling; Mus; Mutate; Neurons; Northern Blotting; Nuclear Localization Signal; Nuclear Structure; Nucleic Acid Regulatory Sequences; Patients; Positioning Attribute; Proteins; Publications; Publishing; Regulation; Role; Skeletal Muscle; Small Interfering RNA; Synapses; Synaptic plasticity; Techniques; Testing; Tg2576; Therapeutic; Thymus Gland; Tissues; Transcript; Variant; Western Blotting; Work; Zinc Fingers; base; brain tissue; disorder control; functional restoration; human tissue; knock-down; mouse model; novel; prevent; promoter; research study; synaptic function; tool; transcription factor

DESCRIPTION (provided by applicant): Accumulating evidence suggests that Alzheimer's Disease (AD) begins as a synaptic disorder. This view is supported by work showing that toxic levels of β-amyloid protein are associated with synaptic dysfunction and memory loss long before atrophy and cell death occur. Inspired by this observation, I have used bioinformatics tools to search for causes of synaptic dysfunction in AD. I have identified a transcription factor called ZCCHC17 that is reduced in AD brain tissue, and my hypothesis is that low levels of this transcription factor contribute to synaptic dysfunction in AD. My hypothesis is supported by my preliminary data (see below). Based on its structure and nuclear localization, ZCCHC17 is predicted to regulate gene expression. However, it is not well studied, and has not previously been linked to AD. Further study of ZCCHC17's role in synaptic dysfunction in AD may provide a therapeutic strategy to rescue synaptic plasticity and memory in this disease. I arrived at my hypothesis using a combination of computational and wet-lab techniques. I began by using novel statistical techniques to screen RNA expression profiles from laser-dissected neurons taken from human AD and control brains. My goal was to identify transcription factors that: 1) Are predicted to regulate a large number of synaptic targets, and 2) Are predicted to have impaired function in AD. My strongest candidate from this work is ZCCHC17. I have now shown that 1) ZCCHC17 protein levels are decreased in AD brain homogenate, and 2) ZCCHC17 is primarily located in neuronal nuclei and is decreased in AD. Although compelling, this data is still only correlational. Since ZCCHC17 is not well studied, the goal of this proposal is to generate foundational data that supports my hypothesis that ZCCHC17 is important for synaptic function and is impaired in AD. This data will justify a more in-depth study of ZCCHC17 in a future project. In Aim 1, I will test the hypothesis that ZCCHC17 is a transcription factor that regulates synaptic genes. I will do this first using ChIP-seq, which will allow me to determine if ZCCHC17 binds to DNA. If ZCCHC17 binds to DNA regulatory regions, then this supports the hypothesis that ZCCHC17 is a transcription factor that regulates gene expression. I will perform ChIP-seq in several different tissues in order to increase my understanding of ZCCHC17's role in neuronal function (human tissue, SY5Y cells, mouse hippocampal neurons, and adult mouse brain). I will also perform siRNA knock-down of ZCCHC17 in the last three tissues. If siRNA knock- down of ZCCHC17 causes reduced expression of ZCCHC17's predicted synaptic targets, then this supports the hypothesis that ZCCHC17 regulates the expression of these synaptic genes. In Aim 2, I will test the hypothesis that elevated β-amyloid levels lead to lower ZCCHC17 protein levels. I have preliminary data supporting this hypothesis, and I will directly test this hypothesis in SY5Y cells, mouse hippocampal neuronal cultures, and in adult mouse cortex. If this prediction is true, then this supports the hypothesis that β-amyloid impairs ZCCHC17 function.

描述（由申请人提供）：越来越多的证据表明阿尔茨海默病（AD）最初是一种突触障碍。这一观点得到了研究的支持，研究表明，早在萎缩和细胞死亡发生之前，β-淀粉样蛋白的毒性水平就与突触功能障碍和记忆丧失有关。受到这一观察结果的启发，我使用生物信息学工具来寻找阿尔茨海默病中突触功能障碍的原因。我已经发现了一种叫做ZCCHC17的转录因子，它在阿尔茨海默氏症的脑组织中减少了，我的假设是这种转录因子的低水平导致了阿尔茨海默氏症的突触功能障碍。我的假设得到了初步数据的支持（见下文）。根据其结构和核定位，预测ZCCHC17具有调控基因表达的功能。然而，它没有得到很好的研究，以前也没有与AD联系起来。进一步研究ZCCHC17在AD突触功能障碍中的作用，可能为挽救该疾病的突触可塑性和记忆提供治疗策略。我结合了计算技术和湿实验室技术得出了我的假设。我开始使用新颖的统计技术从激光解剖的人类AD神经元和控制大脑中筛选RNA表达谱。我的目标是确定转录因子：1)被预测调节大量突触靶点，2)被预测在AD中功能受损。我在这项工作中最强的候选人是ZCCHC17。我现在已经证明，1)ZCCHC17蛋白水平在AD脑匀浆中降低，2)ZCCHC17主要位于神经元核中，在AD中降低。尽管这一数据令人信服，但仍然只是相关性。由于ZCCHC17没有得到很好的研究，本提案的目标是生成基础数据来支持我的假设，即ZCCHC17对突触功能很重要，并且在AD中受损。这些数据将证明在未来的项目中对ZCCHC17进行更深入的研究。在Aim 1中，我将检验ZCCHC17是调节突触基因的转录因子的假设。我将首先使用ChIP-seq，这将使我能够确定ZCCHC17是否与DNA结合。如果ZCCHC17与DNA调控区结合，那么这就支持了ZCCHC17是调节基因表达的转录因子的假设。我将在几个不同的组织（人体组织，SY5Y细胞，小鼠海马神经元，成年小鼠大脑）中进行ChIP-seq，以增加我对ZCCHC17在神经元功能中的作用的了解。我还将在最后三个组织中进行ZCCHC17的siRNA敲低。如果ZCCHC17的siRNA敲低导致ZCCHC17预测的突触靶点的表达减少，那么这就支持了ZCCHC17调节这些突触基因表达的假设。在Aim 2中，我将检验β-淀粉样蛋白水平升高导致ZCCHC17蛋白水平降低的假设。我有支持这一假设的初步数据，我将直接在SY5Y细胞、小鼠海马神经元培养和成年小鼠皮层中验证这一假设。如果这一预测是正确的，那么这就支持了β-淀粉样蛋白损害ZCCHC17功能的假设。