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，阿尔茨海默氏病（AD）开始作为一种突触障碍。这一观点得到了工作的支持，这些工作表明β-淀粉样蛋白的毒性水平与突触功能障碍和记忆丧失有关，早在萎缩和细胞死亡发生之前。受此观察的启发，我使用生物信息学工具来寻找AD中突触功能障碍的原因。我已经确定了一种称为ZCCHC 17的转录因子，它在AD脑组织中减少，我的假设是这种转录因子的低水平有助于AD的突触功能障碍。我的假设得到了初步数据的支持（见下文）。基于其结构和核定位，ZCCHC 17被预测调节基因表达。然而，它并没有得到很好的研究，以前也没有被链接到AD。进一步研究ZCCHC 17在AD突触功能障碍中的作用可能为挽救这种疾病的突触可塑性和记忆提供治疗策略。我通过计算和湿实验室技术的结合得出了我的假设。首先，我使用新的统计技术，从AD患者和对照组大脑的激光切割神经元中筛选RNA表达谱。我的目标是确定以下转录因子：1）预测调节大量突触靶点，2）预测在AD中功能受损。我在这项工作中最强的候选人是ZCCHC 17。我现在已经表明，1）ZCCHC 17蛋白水平在AD脑匀浆中降低，2）ZCCHC 17主要位于神经元核中，并在AD中降低。尽管这些数据很有说服力，但它们仍然只是相互关联的。由于ZCCHC 17还没有得到很好的研究，本提案的目标是生成支持我的假设的基础数据，即ZCCHC 17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ZCCHC 17进行更深入的研究是合理的。在目标1中，我将检验ZCCHC 17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ZCCHC 17是否与DNA结合。如果ZCCHC 17与DNA调控区结合，则支持ZCCHC 17是调控基因表达的转录因子的假设。我将在几种不同的组织中进行ChIP-seq，以增加我对ZCCHC 17在神经元功能中作用的理解（人体组织，SY 5 Y细胞，小鼠海马神经元和成年小鼠大脑）。我还将在最后三个组织中进行ZCCHC 17的siRNA敲低。如果ZCCHC 17的siRNA敲低导致ZCCHC 17预测的突触靶的表达减少，则这支持ZCCHC 17调节这些突触基因的表达的假设。在目标2中，我将检验β-淀粉样蛋白水平升高导致ZCCHC 17蛋白水平降低的假设。我有初步的数据支持这一假设，我将在SY 5 Y细胞、小鼠海马神经元培养物和成年小鼠皮层中直接检验这一假设。如果这一预测是正确的，那么这支持了β-淀粉样蛋白损害ZCCHC 17功能的假设。