STUDYING THE ROLE OF INFLAMMATORY MEDIATOR MIR-155 IN A MOUSE MODEL OF ALS

研究炎症介质 MIR-155 在 ALS 小鼠模型中的作用

• 批准号: 8489135
• 负责人: Erica Danielle Koval
• 金额: $ 2.86万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-07-01 至 2014-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8489135
• 关键词: A Mouse; Ablation; Adult; Affect; Age; Amyotrophic Lateral Sclerosis; Antisense Oligonucleotides; Atrophic; Autopsy; B-Lymphocytes; Binding Sites; Biological Assay; Biological Process; Brain; Breeding; Cells; Cerebrospinal Fluid; Cessation of life; Complement; Data; Development; Disease; Disease Pathway; Disease Progression; Drug Delivery Systems; Familial Amyotrophic Lateral Sclerosis; Future; Gene Mutation; Human; Human Genetics; Immune; Immune Cell Activation; In Situ Hybridization; Inflammation; Inflammation Mediators; Inflammatory; Intervention; Knock-out; Learning; Link; Longevity; Mediating; Messenger RNA; MicroRNAs; Microglia; Modeling; Modification; Molecular; Motor Neurons; Mus; Muscle Weakness; Nerve Degeneration; Neurodegenerative Disorders; Neuromuscular Diseases; Onset of illness; Pathogenesis; Pathway interactions; Patients; Peripheral; Phenocopy; Phenotype; Proteins; Pump; RNA; Research; Rodent Control; Rodent Model; Role; Severity of illness; Spinal Cord; T-Lymphocyte; Techniques; Technology; Testing; Therapeutic; Time; Tissues; Translational Research; Western Blotting; Work; cytokine; derepression; design; disease phenotype; experience; in vivo; inhibitor/antagonist; insight; interest; knock-down; knockout animal; macrophage; mouse model; nervous system disorder; neuroinflammation; novel; research study; screening; tool

DESCRIPTION (provided by applicant): Amyotrophic lateral sclerosis (ALS) is a fatal, adult-onset neuromuscular disorder characterized by the progressive loss of motor neurons in the spinal cord and brain. There is no adequate therapeutic for ALS, and patients typically die within 5 years of disease onset1. microRNAs are a promising new avenue for disease research. microRNAs are short single-stranded RNA molecules capable of regulating large numbers of mRNAs. microRNAs are dysregulated in a variety of diseases and have proven themselves to be viable therapeutic targets20,21. By comparing microRNAs in the spinal cords of our ALS rodent model to age-matched controls, we discovered a robust and significant increase in specific microRNAs. Of particular interest is miR-155, a microRNA expressed by immune cells and known to be involved in inflammation and immunity27. We have since confirmed this microRNA to also be significantly upregulated in ALS patient autopsy spinal cord tissue. We will study the role of this inflammatory mediating microRNA in our ALS rodent model in order to elucidate its role in neurodegeneration and to test its intervention potential. In Aim 1, we will collect time course data and perform in situ hybridization in order to learn when the expression levels of our microRNA of interest begin to change and in order to determine which cells in the spinal cord dynamically modulate their miR-155 expression during disease. Next, in Aim 2, we will determine the importance of miR-155 during neurodegenerative disease and its impact on disease pathways. We are currently breeding miR-155 knockout animals into our ALS mouse line. We will compare the mice in their age of disease onset and overall longevity. In order to link mechanism to phenotype, we will complement these studies with immunohistological and molecular assays that will evaluate changes in neuroinflammatory pathways. Finally, in Aim 3, we will knock down miR-155 with antisense oligonucleotides delivered directly into the cerebral spinal fluid with osmotic pump technology. In addition to developing a novel knockdown technique for microRNAs, this work will also allow us to modulate our microRNA both temporally and spatially - furthering our mechanistic insight (peripheral vs. central effect) while also testig a potential therapeutic. If successful, the proposed experiments will expand our understanding of the role of microRNAs in neuroinflammation and may also help inform the development of a novel ALS therapeutic.

描述（由申请人提供）：肌萎缩性侧索硬化症（ALS）是一种致命的、成人发病的神经肌肉疾病，其特征在于脊髓和大脑中运动神经元的进行性丧失。ALS没有足够的治疗方法，患者通常在疾病发作后5年内死亡1。microRNA是疾病研究的一个有前途的新途径。microRNA是能够调节大量mRNA的短单链RNA分子。microRNA在多种疾病中失调，并且已经证明它们本身是可行的治疗靶点20，21。通过比较我们的ALS啮齿动物模型脊髓中的microRNA与年龄匹配的对照组，我们发现特定microRNA的稳健和显着增加。特别令人感兴趣的是miR-155，一种由免疫细胞表达的微小RNA，已知参与炎症和免疫27。我们已经证实这种microRNA在ALS患者尸检脊髓组织中也显著上调。我们将研究这种炎症介导的microRNA在ALS啮齿动物模型中的作用，以阐明其在神经退行性变中的作用并测试其干预潜力。在目标1中，我们将收集时间进程数据并进行原位杂交，以了解我们感兴趣的microRNA的表达水平何时开始变化，并确定脊髓中哪些细胞在疾病期间动态调节其miR-155表达。接下来，在目标2中，我们将确定miR-155在神经退行性疾病中的重要性及其对疾病途径的影响。我们目前正在将miR-155敲除动物培育成我们的ALS小鼠系。我们将比较小鼠的发病年龄和总体寿命。为了将机制与表型联系起来，我们将用免疫组织学和分子测定来补充这些研究，以评估神经炎症通路的变化。最后，在目标3中，我们将使用渗透泵技术直接递送到脑脊液中的反义寡核苷酸敲除miR-155。除了为microRNA开发一种新的敲低技术外，这项工作还将使我们能够在时间和空间上调节我们的microRNA-促进我们的机制洞察力（外周效应与中枢效应），同时也测试了潜在的治疗方法。如果成功，拟议的实验将扩大我们对microRNA在神经炎症中作用的理解，也可能有助于开发一种新的ALS治疗方法。