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（由适用提供）：积累证据表明阿尔茨海默氏病（AD）始于合成疾病。这种观点得到的工作表明，β-淀粉样蛋白的毒性水平与合成功能障碍和记忆丧失有关，早在萎缩和细胞死亡发生之前。受到这一观察的启发，我使用了生物信息学工具来寻找AD中合成功能障碍的原因。我已经确定了一种称为ZCCHC17的转录因子，该转录因子在AD脑组织中降低，我的假设是，该转录因子的低水平导致AD突触功能障碍。我的假设得到了我的初步数据的支持（见下文）。根据其结构和核定位，ZCCHC17预计将调节基因表达。但是，它不是很好的研究，并且以前尚未与AD联系在一起。研究ZCCHC17在AD中突触功能障碍中的作用可能提供了一种治疗策略来挽救该疾病中的合成可塑性和记忆力。我结合了计算和湿lab技术的结合，得出了假设。我首先使用新颖的统计技术来筛选从人类AD和对照大脑中删除的激光神经元的RNA表达谱。我的目标是确定：1​​）预计将调节大量突触靶标的转录因子，而2）预计在AD中的功能受损。我从这项工作中的有力候选人是ZCCHC17。我现在已经表明，1）ZCCHC17蛋白水平在AD脑匀浆中降低，而2）ZCCHC17主要位于神经元核中，AD中的AD蛋白水平降低。尽管令人信服，但这些数据仍然仅相关。由于ZCCHC17的研究不是很好，因此该建议的目的是生成支持我的基础数据，该数据支持我的假设，即ZCCHC17对于突触功能很重要，并且在AD中受到了损害。这些数据将证明对未来项目中ZCCHC17进行更深入的研究是合理的。在AIM 1中，我将检验以下假设：ZCCHC17是调节突触基因的转录因子。我将首先使用ChIP-Seq进行此操作，这将使我能够确定ZCCHC17是否与DNA结合。如果ZCCHC17与DNA调节区域结合，则支持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中执行CHIP-SEQ，以提高我对ZCCHC17在神经元功能（人体组织，SY5Y细胞，小鼠海马神经元和成年小鼠脑的作用）的理解。我还将在最后三个组织中进行ZCCHC17的siRNA敲除。如果将ZCCHC17的siRNA敲除导致ZCCHC17预测的突触靶标的表达降低，那么这支持了ZCCHC17调节这些合成基因表达的假设。在AIM 2中，我将测试以下假设：升高的β-淀粉样蛋白水平导致ZCCHC17蛋白水平降低。我有支持这一假设的初步数据，并将直接在SY5Y细胞，小鼠海马神经元培养物和成年小鼠皮层中直接检验该假设。如果此预测是正确的，则支持β-淀粉样蛋白会损害ZCCHC17功能的假设。