Neurofilaments, SOD1 and Motor Neuron Diseases

神经丝、SOD1 和运动神经元疾病

• 批准号: 8656149
• 负责人: Don W Cleveland
• 金额: $ 48.76万
• 依托单位: LUDWIG INSTITUTE FOR CANCER RES LTD
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-04-01 至 2017-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8656149
• 关键词: Affect; Affinity; Amyotrophic Lateral Sclerosis; Antisense Oligonucleotides; Astrocytes; Automobile Driving; Autopsy; CSPG4 gene; Cells; Cessation of life; Complement; Complement 1q; Cytoplasm; Disease; Disease Progression; Event; Familial Amyotrophic Lateral Sclerosis; Genes; Genetic; High-Throughput Nucleotide Sequencing; Immunofluorescence Immunologic; Immunoprecipitation; Infusion procedures; Inherited; Link; MIF gene; Mass Spectrum Analysis; Measures; Mediating; Messenger RNA; Microglia; Mitochondria; Mitochondrial Proteins; Modeling; Molecular Chaperones; Motor Neuron Disease; Motor Neurons; Mus; Mutant Strains Mice; Mutation; Neuroglia; Neurons; Oligodendroglia; Onset of illness; Paralysed; Pathogenesis; Patients; Prevalence; Protein Import; Proteins; Role; Schwann Cells; Solutions; Spinal Cord; Superoxide Dismutase; Synapses; Testing; Tissues; Toxic effect; Transgenes; cell type; interest; mutant; neural precursor cell; neurofilament; precursor cell; premature; recombinase; selective expression

DESCRIPTION (provided by applicant): A proportion of dominantly inherited ALS arises from mutation in superoxide dismutase (SOD1). Accumulation of misfolded SOD1 is widely recognized as a component of this toxicity, especially its aggregation onto mitochondria within spinal cord. How mitochondrial composition is affected by mutant SOD1 will be determined using quantitative SILAM mass spectrometry. The mechanism(s) through which ALS-linked mutations aggregate and damage mitochondria only in affected tissues will be also be determined, focusing on our discovery of a chaperone that can block misfolded SOD1 accumulation in non-neuronal cells. Combining 1) Barres' discovery of a role for complement in synaptic pruning and 2) our discovery that components of the complement cascade are induced in motor neurons early in SOD1 mutant-mediated disease, gene disruption will now be used to test the role in disease pathogenesis of complement induction within motor neurons. We previously demonstrated that toxicity from SOD1 mutants is non-cell autonomous, with damage within motor neurons driving disease onset and damage within neighboring glial cells (both astrocytes and microglia) driving rapid disease progression. The contribution(s) of mutant SOD1 toxicity within additional cell types, especially oligodendrocytes and their precursors will be tested by deletion of the mutant encoding transgene using cell type specific expression of Cre recombinase. Mechanistically, how mutant SOD1 damages motor neurons, astrocytes and oligodendrocytes will be identified by high throughput sequencing of polysomal mRNAs recovered by ribosomal affinity tagging. This question is of especially high interest for astrocytes, which are known to generate one or more toxicities from their synthesis of ALS causing mutants in SOD1.

描述（由申请人提供）：一部分显性遗传性ALS由超氧化物歧化酶（SOD 1）突变引起。错误折叠的SOD 1的积累被广泛认为是这种毒性的一个组成部分，特别是它在脊髓内的线粒体上的聚集。将使用定量SILAM质谱法来确定突变SOD 1如何影响线粒体组成。ALS连锁突变仅在受影响的组织中聚集和损伤线粒体的机制也将被确定，重点是我们发现了一种可以阻断非神经元细胞中错误折叠的SOD 1积累的分子伴侣。结合1）Barres关于补体在突触修剪中的作用的发现和2）我们关于补体级联的组分在SOD1突变体介导的疾病早期在运动神经元中被诱导的发现，基因破坏现在将用于测试运动神经元内补体诱导在疾病发病机制中的作用。我们以前证明，SOD1突变体的毒性是非细胞自主的，运动神经元内的损伤驱动疾病发作，相邻神经胶质细胞（星形胶质细胞和小胶质细胞）内的损伤驱动疾病快速进展。通过使用Cre重组酶的细胞类型特异性表达缺失编码转基因的突变体来测试突变体SOD 1毒性在其他细胞类型，特别是少突胶质细胞及其前体中的贡献。从机制上讲，突变体SOD1如何损伤运动神经元、星形胶质细胞和少突胶质细胞将通过核糖体亲和标记回收的多核糖体mRNA的高通量测序来鉴定。这个问题对于星形胶质细胞特别重要，已知星形胶质细胞从其合成导致SOD1突变体的ALS中产生一种或多种毒性。