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.

酗酒会对大脑和行为产生毁灭性的影响，并对公众健康造成巨大负担。 然而，酒精所致神经细胞损伤的分子基础尚不完全清楚。近期 功能基因组学的进展表明，长的非编码RNA(LncRNAs)可能在酒精- 诱导神经细胞死亡。我们实验室最近的工作支持了这一假设。因此，a 建立酒精性神经细胞死亡相关基因的综合筛选体系具有重要意义 确定潜在治疗靶点的好处。 基因编辑工具，如锌指核酸酶(ZFN)和转录激活样元件 核酸酶(TALEN)在基因功能研究或基因治疗应用中具有巨大的潜力 通过击倒或击倒技术。一种基于细菌聚类的新型基因编辑工具 规则间隔短回文重复序列(CRISPR)/CRISPR相关系统[CAS])已被 成功地应用于从细菌、酵母到哺乳动物的各种生物中。相对于ZFN和TALEN， CRISPR/Cas是有利的，因为它只需要改变引导RNA(GRNA)的序列，并且它 也可以直接输送到胚胎中，产生基因修饰的生物。此外，多路传输 CRISPR/Cas的能力使得同时靶向多个基因成为可能。 在这项研究中，我们将使用我们最近开发的双引导CRISPR/CAS方法来产生lncRNA SH-SY5Y细胞模型中的基因敲除(GRNA)文库。然后我们将对lncRNAs进行全基因组筛选 参与酒精诱导的细胞死亡途径。我们的初步数据表明，酒精会增加 核旁斑点蛋白基因在SH-SY5Y细胞中的表达我们之前的工作已经证明 酒精以剂量依赖的方式激活氧化应激-凋亡细胞死亡途径，以减少 神经细胞活性和增加细胞氧化应激。我们期待着高吞吐量的应用 酒精诱导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lncRNAs及其在介导酒精诱导的神经变性中的潜在作用。生成的 本研究中的lncRNA gRNA文库也可用于表征其他化合物诱导的细胞毒性 神经毒素或其他物质滥用。因此，这项研究有可能减少酒精对健康的影响 对滥用药物和其他与公共卫生负担有关的物质滥用也有好处。