Long non-coding RNAs in alcohol induced neural cell death

酒精诱导的神经细胞死亡中的长非编码RNA

• 批准号: 9386466
• 负责人: YIN-YUAN MO
• 金额: $ 22.28万
• 依托单位: UNIVERSITY OF MISSISSIPPI MED CTR
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-07-20 至 2019-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9386466
• 关键词: Affect; Alcohol abuse; Alcohol consumption; Alcohol-Induced Neurotoxicity; Alcohols; Animals; Apoptotic; Atrophic; Bacteria; Brain Injuries; Cell Death; Cell Line; Cell Nucleus; Cell Survival; Cell model; Cell physiology; Cells; Clustered Regularly Interspaced Short Palindromic Repeats; Code; Data; Dose; Elements; Embryo; Gene Expression; Gene-Modified; Genes; Genetic; Genetic Transcription; Genome; Goals; Guide RNA; Health; Hippocampus (Brain); Human; Individual; Knock-in; Knock-out; Laboratories; Libraries; Mammals; Mediating; MicroRNAs; Molecular; Molecular Target; Nerve Degeneration; Neurons; Neurotoxins; Nuclear; Occupations; Organism; Outcomes Research; Oxidative Stress; Pathway interactions; Play; Prefrontal Cortex; Proteins; Public Health; RNA library; Research; Resistance; Risk; Rodent; Role; Sampling; Series; Signal Transduction; Site; Source; Structure; Study models; Substance abuse problem; System; Techniques; Testing; Toxic effect; Transcript; Transcriptional Activation; Untranslated RNA; Validation; Work; Yeasts; alcohol abuse therapy; alcohol response; alcohol use disorder; base; biological adaptation to stress; brain behavior; brain cell; cell injury; chronic alcohol ingestion; cognitive ability; deep sequencing; dentate gyrus; experimental study; functional genomics; gene therapy; genome-wide; genome-wide analysis; high throughput screening; in vitro Model; neuron loss; neurotoxicity; nuclease; particle; prevent; screening; therapeutic development; therapeutic target; tool; zinc finger nuclease

Alcohol abuse can have devastating impacts on the brain and behavior and is a big burden on public health. However, the molecular underpinnings of alcohol induced neural cell injury are not fully understood. Recent advances in functional genomics suggest that long non-coding RNAs (lncRNAs) may play critical roles in alcohol- induced neural cell death. The recent work from our laboratory support this hypothesis. Therefore, a comprehensive screening system of lncRNAs related to alcohol-induced neural cell death is of significant benefits for identifying potential therapeutic targets. Genetic editing tools such as Zinc Finger Nuclease (ZFN) and Transcription Activation-Like Element Nuclease (TALEN) have great potential for the functional study of genes or the application of gene therapy through knockout or knockin techniques. A new type of genetic editing tool based on bacterial Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/CRISPR‐Associated System [Cas]) has been successfully used in various organisms from bacteria and yeast to mammals. Relative to ZFN and TALEN, CRISPR/Cas is advantageous because it only requires changing the sequence of the guide RNA (gRNA) and it can also be directly delivered into embryos to generate gene-modified organisms. Furthermore, the multiplexing capability of CRISPR/Cas makes it possible to target multiple genes simultaneously. In this study we will use our recently developed dual guide CRISPR/Cas approach to generate an lncRNA knockout (gRNA) library in SH-SY5Y cell model. We will then perform a genome-wide screening for lncRNAs involved in alcohol-induced cell death pathways. Our preliminary data indicated that alcohol increased the expression of a nuclear paraspeckle lncRNA in SH-SY5Y cells. Our previous work have demonstrated that alcohol activates the oxidative stress-apoptotic cell death pathway in a dose dependent manner to reduce neuronal cell viability and increase cellular oxidative stress. We expect that the application of the high throughput screening platform in alcohol-induced neuronal death system will allow for the accurate identification of all other specific lncRNAs as well as their potential roles in mediating alcohol-induced neurodegeneration. The generated lncRNA gRNA library in this study can also be available for the characterization of cell toxicity induced by other neurotoxins or other substance abuse. Thus this research has a potential to reduce the health impacts of alcohol abuse and also has benefits for other substances abuse related public heath burdens.

酗酒可能会对大脑和行为产生毁灭性的影响，这对公共卫生有很大的影响。 然而，酒精诱导的神经细胞损伤的分子基础尚未完全了解。最近的 功能基因组学的进步表明，长的非编码RNA（LNCRNA）可能在酒精中起关键作用 - 诱导神经元细胞死亡。我们实验室的最新工作支持这一假设。因此， 与酒精引起的神经细胞死亡有关的LNCRNA的综合筛查系统是显着的 确定潜在的治疗靶标的好处。 遗传编辑工具，例如锌指核酸酶（ZFN）和转录激活元件 核酸酶（TALEN）具有基因功能研究或基因治疗的巨大潜力 通过敲除或敲门技术。基于细菌聚类的新型遗传编辑工具 定期间隔短的短质体重复序列（CRISPR）/CRISPR相关系统[CAS]） 从细菌和酵母到哺乳动物的各种生物中成功使用。相对于ZFN和TALEN， CRISPR/CAS是有利的，因为它仅需要更改导向RNA（GRNA）的顺序，并且它 也可以直接输送到胚胎中以产生基因修饰的生物。此外，多路复用 CRISPR/CAS的能力使得可以简单地靶向多个基因。 在这项研究中，我们将使用我们最近开发的双指南CRISPR/CAS方法来生成lncrna SH-SY5Y细胞模型中的敲除（GRNA）库。然后，我们将对LNCRNA进行全基因组筛查 参与酒精引起的细胞死亡途径。我们的初步数据表明酒精增加了 SH-SY5Y细胞中的核parpecckle lncRNA的表达。我们以前的工作证明了 酒精以剂量依赖性的方式激活氧化应激 - 凋亡细胞死亡途径 神经元细胞活力并增加细胞氧化物应激。我们期望高通量的应用 酒精引起的神经元死亡系统中的筛查平台将允许准确识别所有其他 特定的LNCRNA及其在介导酒精诱导的神经退行性中的潜在作用。生成的 本研究中的lncRNA grna文库也可以用于表征其他细胞毒性 神经毒素或其他滥用药物。这项研究有可能减少酒精的健康影响 滥用，还为其他与滥用药物有关的公共荒地伯伦斯带来了好处。