Novel ALS models: FUS conditional knockout in motor neurons and oligodendrocytes

新型 ALS 模型：运动神经元和少突胶质细胞中的 FUS 条件敲除

• 批准号: 9026836
• 负责人: FRANCA CAMBI
• 金额: --
• 依托单位: VETERANS HEALTH ADMINISTRATION
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-01-01 至 2017-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9026836
• 关键词: Accounting; Adult; Affect; Amyotrophic Lateral Sclerosis; Area; Award; Breathing; Cells; Clinical Trials; Cytoplasm; Cytoplasmic Granules; DNA; Data; Defect; Development; Disease; Drosophila genus; Functional disorder; Funding; Future; Goals; Health Benefit; Human; Inherited; Intervention; Knock-out; Knockout Mice; Knowledge; Laboratories; Male Sterility; Mediating; Metabolism; Modeling; Molecular; Molecular Target; Motor; Motor Neuron Disease; Motor Neurons; Mus; Mutation; Neonatal; Nerve Degeneration; Nervous system structure; Neuroglia; Neurons; Nuclear; Nuclear Export; Nuclear Localization Signal; Oligodendroglia; Phenotype; Pilot Projects; Play; Process; Processed Genes; Proteins; RNA; Reagent; Research; Research Personnel; Role; Signal Transduction; Spinal Cord; Stress; Supporting Cell; Testing; Veterans; Zebrafish; age related; base; disease-causing mutation; effective therapy; falls; gain of function; gene discovery; in vivo; innovation; loss of function; motor neuron degeneration; mouse model; mutant; neuron loss; neurotoxicity; novel; novel therapeutic intervention; novel therapeutics; overexpression; public health relevance; sarcoma

 DESCRIPTION (provided by applicant): Amyotrophic Lateral Sclerosis (ALS) is an invariably fatal disorder caused by degeneration of the upper and lower motor neuron. Presently, there are no effective therapies. Many promising interventions have failed in clinical trials. New therapies are needed and their development will depend on advances in our knowledge of mechanisms of motor neuron degeneration and of the support provided by the surrounding glia. Fused in sarcoma (FUS) is one of the latest genes discovered to cause familial and sporadic ALS. It is one of a growing number of RNA/DNA processing genes associated with familial and sporadic ALS. This discovery has further highlighted the pivotal role that defects in RNA/DNA function and processing have in neurodegeneration. A major unanswered question is whether loss of FUS function contributes to neuronal degeneration. Ours and other investigators studies support the notion that loss of nuclear function(s) plays a role in motor neuron degeneration, however, the importance of loss of function mechanism in vivo remains to be determined. Another critical question is whether neurodegeneration caused by FUS defects is cell-autonomous. Our studies in Drosophila suggest a glia-based mechanism of neurotoxicity. The hypothesis to be tested in this project is that loss of FUS function is a critical mechanism to cause motor neuron degeneration and non cell-autonomous FUS neurotoxicity mediated by oligodendrocytes plays an important role in ALS. Constitutive FUS knockout in mice resulted in neonatal lethality and male sterility, hence these mice cannot be used to investigate the role of loss of FUS in vivo in the adult nervous system. To fill this gap in knowledge, we will make FUS conditional knockout mice in which FUS is depleted in either motor neurons or oligodendrocytes and characterize age-dependent motor neuron loss and associated phenotypes. We have made a FUSfl/fl mouse line and we are ready to cross it with neuronal and oligodendrocyte Cre expressing lines. We will perform motor, pathological and molecular studies to determine whether FUS deficient motor neurons undergo age-dependent degeneration and whether FUS deficient oligodendrocytes contribute to motor neuron loss. Future studies will investigate the molecular mechanisms of disease in these two models. The ultimate goal is to generate models in which molecular targets can be identified, characterized and used to test novel therapeutic interventions for ALS. Collectively, these studies will fill an important gap in knowledge and will generate valuable mouse models that will be used to investigate disease mechanisms and serve as in vivo platforms to test compounds aimed at slowing progression of motor neuron loss. These studies have a great potential to generate results and reagents that will advance the field of ALS research and will benefit the health of Veterans with motor neuron disorders.

 描述（由申请人提供）： 肌萎缩性侧索硬化症（ALS）是由上和下运动神经元变性引起的致命性疾病。目前，没有有效的疗法。在临床试验中，许多承诺干预措施失败了。需要新的疗法，它们的发育将取决于我们对运动神经元变性机理的了解以及周围神经胶质的支持。在肉瘤中融合（FUS）是发现家庭和零星ALS的最新基因之一。它是越来越多的与家族和零星ALS相关的RNA/DNA加工基因之一。这一发现进一步强调了RNA/DNA功能中缺陷的关键作用，并且在神经变性中具有处理。一个主要的未解决的问题是FUS功能的丧失是否有助于神经变性。我们的和其他研究人员的研究支持这样的观念：核功能丧失在运动神经变性中起作用，但是，体内功能机制丧失的重要性仍然有待确定。另一个关键的问题是，由FUS缺陷引起的神经退行性是否是细胞自主的。我们对果蝇的研究提出了基于神经胶质的神经毒性机制。该项目中要检验的假设是，FUS功能的丧失是引起运动神经元变性和由少突胶质细胞介导的非细胞自主FUS神经毒性的关键机制，在ALS中起重要作用。小鼠的组成型FUS敲除导致新生儿致死性和雄性不育，因此这些小鼠不能用于研究成人神经系统中FUS损失的作用。为了填补知识的空白，我们将使有条件的敲除小鼠在运动神经元或少突胶质细胞中耗尽FU的小鼠，并表征依赖年龄的运动神经元损失和相关表型。我们已经制作了FUSFL/FL小鼠系，我们准备用神经元和少突胶质细胞表达线横穿它。我们将进行运动，病理和分子研究，以确定FUS确定性运动神经元是否经历年龄依赖性变性，以及FUS确定的少突胶质细胞是否有助于运动神经元丧失。未来的研究将研究这两个模型中疾病的分子机制。最终目标是生成模型，在该模型中，可以将分子靶标识别，表征和用于测试ALS的新型治疗干预措施。总的来说，这些研究将填补知识的重要空白，并将产生有价值的小鼠模型，这些模型将用于研究疾病机制，并用作测试旨在减慢运动神经元损失进展的化合物的体内平台。这些研究具有产生结果和试剂的巨大潜力，可以推进ALS研究领域，并将使退伍军人患有运动神经元疾病的健康受益。