ANALYZING MICROGLIA-DERIVED TOXICITY TO MOTOR NEURONS IN ALS

分析 ALS 中小胶质细胞对运动神经元的毒性

• 批准号: 8171449
• 负责人: Don W Cleveland
• 金额: $ 0.24万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8171449
• 关键词: Amyotrophic Lateral Sclerosis; Cells; Cessation of life; Computer Retrieval of Information on Scientific Projects Database; Conditioned Culture Media; Cuprozinc Superoxide Dismutase; Disease; Disease Progression; Familial Amyotrophic Lateral Sclerosis; Funding; Genes; Grant; Immune; Institution; Mass Spectrum Analysis; Microglia; Motor Neurons; Mus; Mutant Strains Mice; Mutation; Neurodegenerative Disorders; Neuroglia; Neurons; Paralysed; Pathway interactions; Research; Research Personnel; Resources; Source; Toxic effect; Transgenic Mice; United States National Institutes of Health; cell type; central nervous system injury; macrophage; mutant; release factor

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Amyotrophic Lateral Sclerosis (ALS) is a late onset neurodegenerative disease leading to paralysis. Mutations in the gene for the ubiquitously expressed Cu/Zn superoxide dismutase (SOD1) are the best-known cause for familial ALS and transgenic mice constitutively expressing mutant SOD1 develop a late-onset, ALS-like disease. It is known that degeneration of upper and lower motor neurons is responsible for paralysis in ALS and that glial cell types expressing mutant SOD1 contribute to disease mechanism. We showed that microglial cells, the immune cells of the CNS, were important players, since diminishing mutant SOD1 specifically in macrophages/ microglial cells extended survival in ALS mice. Microglial cells are activated in any injury of the CNS including sporadic and familial ALS and when activated, they release factors that can be toxic or trophic for neurons. Therefore, identifying those factors could be a key to finding new targets implicated in motor neuron death. As downregulating mutant SOD1 from macrophages/microglial cells slowed disease progression in ALS mice, mutant SOD1 must act directly within microglial cells to generate toxicity toward motor neurons. Therefore, microglial cells expressing mutant or wild-type SOD1 will be screened for the different factors that they can release using Mass Spectrometry. Conditioned medium from cultured microglial cells will be used as a source of released microglial factors and medium from cells expressing mutant SOD1 will be compared to medium from wild-type SOD1 expressing microglial cells. Identification of the differences between mutant SOD1 expressing microglial cells and control microglial cells should help elucidate the intracellular pathways involved and the toxic factors released by microglial cells that contribute to motor neuron death.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 肌萎缩侧索硬化症（ALS）是一种迟发性神经退行性疾病，可导致瘫痪。普遍表达的铜/锌超氧化物歧化酶 (SOD1) 基因突变是导致家族性 ALS 的最常见原因，而持续表达突变 SOD1 的转基因小鼠会患上迟发性 ALS 样疾病。众所周知，上运动神经元和下运动神经元的变性是 ALS 瘫痪的原因，而表达突变型 SOD1 的神经胶质细胞类型则有助于疾病机制。我们发现小胶质细胞（中枢神经系统的免疫细胞）是重要的参与者，因为减少巨噬细胞/小胶质细胞中的突变型 SOD1 可延长 ALS 小鼠的存活时间。小胶质细胞在中枢神经系统的任​​何损伤（包括散发性和家族性 ALS）中都会被激活，当激活时，它们会释放对神经元有毒或有营养的因子。因此，识别这些因素可能是寻找与运动神经元死亡有关的新靶点的关键。由于下调巨噬细胞/小胶质细胞的突变型 SOD1 可以减缓 ALS 小鼠的疾病进展，因此突变型 SOD1 必须直接在小胶质细胞内发挥作用，对运动神经元产生毒性。因此，将使用质谱法筛选表达突变型或野生型 SOD1 的小胶质细胞，以确定它们可以释放的不同因子。来自培养的小胶质细胞的条件培养基将用作释放的小胶质细胞因子的来源，并且来自表达突变体SOD1的细胞的培养基将与来自表达野生型SOD1的小胶质细胞的培养基进行比较。鉴定表达突变型 SOD1 的小胶质细胞和对照小胶质细胞之间的差异应有助于阐明所涉及的细胞内途径以及小胶质细胞释放的导致运动神经元死亡的毒性因子。