Mechanisms of Protection and Pathogenesis in Amyotrophic Lateral Sclerosis Mice

肌萎缩侧索硬化症小鼠的保护机制和发病机制

基本信息

批准号：
7260026
负责人：
JOHN P CROW
金额：
$ 38.11万
依托单位：
UNIV OF ARKANSAS FOR MED SCIS
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-12-01 至 2012-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7260026
关键词：
2-Mercaptoethanol Acids Active Sites Address Affect Affinity Age Agreement Alkylation Amino Acid Substitution Amyotrophic Lateral Sclerosis Animals Antibodies Applications Grants Back Binding Binding Sites Biological Markers Brain California Cell Death Process Cells Cessation of life Characteristics Chemicals Clinical Clinical Trials Condition Conflict (Psychology)Copper Core Facility Crows Cysteine Cytosol Data Dependence Detection Disease Disease Progression Disruption Disulfides Drug Controls Drug Design Elements Enzyme Stability Enzymes Event Evolution Experimental Designs Failure Fluorescence Funding Future Gel Gene Expression Genes Genetic Crosses Goals Grant Heart High Pressure Liquid Chromatography Histidine Histopathology Human Immunoblotting In Vitro Injury Investigation Isotopes Knock-out Knowledge Laboratories Lead Life Extension Light Link Localized Location Longevity Mass Spectrum Analysis Measurement Mediating Membrane Messenger RNA Metals Methods Microglia Microscopic Mitochondria Modification Molecular Molecular Chaperones Molecular Weight Monitor Morphologic artifacts Motor Neuron Disease Motor Neurons Mus Mutation Neuroglia Neuronal Injury Neurons Numbers Object Attachment One-Step dentin bonding system Onset of illness Outcome Pathogenesis Pathway interactions Pattern Peptides Peroxonitrite Pharmaceutical Preparations Phase Phenotype Phenylalanine Play Positioning Attribute Post-Translational Protein Processing Preparation Principal Investigator Process Progress Reports Property Protein Binding Protein Overexpression Proteins Proteomics Protocols documentation Publications Published Comment Publishing Rate Rattus Reaction Reader Recombinant Proteins Recommendation Relative (related person)Reporting Research Design Research Personnel Resistance Role Running Sampling Scanning Scheme Secondary to Severity of illness Shoulder Signal Transduction Site Small Interfering RNA Solvents Source Spinal Cord Spottings Staging Sulfhydryl Compounds Superoxides Surface Symptoms System Techniques Technology Testing Text Therapeutic Therapeutic Effect Therapeutic Uses Time Tissues Toxic effect Transgenic Mice Transgenic Organisms Translating Treatment Protocols Tryptophan Uncertainty Update Urea Weight Work Zinc Zinc deficiency absorption analog base chemical property concept crosslink day design dimer disulfide bond drug development gel electrophoresis gene therapy high throughput analysis hydroxy-aluminum polymer in vivo innovation insight interest killings knock-down laser capture microdissection link protein molecular mass monomer mouse model mutant novel oxidation persulfides polypeptide prevent programs protective effect protein crosslink protein expression protein folding protein misfolding ranpirnase research study residence scale up uptake

项目摘要

DESCRIPTION (Provided by applicant): For over a decade, ALS-associated SOD1 mutants have been compared to wild-type enzyme, both in vitro and in vivo in an attempt to understand the mechanism(s) of pathogenesis. However, little is known about the molecular mechanisms which underlie pharmacologically-mediated extension of survival in G93A mice. We have identified two chemically distinct (organomanganic) compounds which markedly extend survival in G93A mice when given at disease onset (similar to when human treatment would begin). Based on our results, efforts are underway to conduct human clinical trials in ALS with these compounds. We propose to use these highly efficacious compounds to probe the mechanisms of protection, while continuing our work on the mechanisms of SOD1 mutant-mediated toxicity. We will examine protein expression changes which accompany protection by these compounds and the ultimate loss of protection (Aim 1) in order to gain a better understanding of the mechanisms involved. The magnitude of life extension with these compounds rivals the best yet reported for any compound, even those administered presymptomatically. However, reliable comparisons require comparable treatment regimens. We propose to test compounds that show efficacy when given presymptomatically in an onset administration paradigm (Aim 2) to test the hypothesis that the failure of some human trials may relate to differences in study design. Compelling new evidence suggests that mitochondrial uptake of mutants is critical to disease and that uptake is determined by metallation state (primarily zinc) and the status of an internal disulfide. Herein we offer fundamentally new evidence that a zinc-deficient mutant in vivo directly correlates with disease severity, and that mutant SOD1 (but not wild-type) is covalently cross linked. Aim 3 will identify the cross linked proteins and determine if drug treatment alters it. Aim 4 will extend our studies of in vivo metallation states and thiol status of mutants using enzyme purified from spinal cord as an "in vivo expression system".

描述（由申请人提供）：十多年来，将与ALS相关的SOD1突变体与野生型酶进行了比较，无论是在体外还是体内，都试图了解发病机理的机制。然而，对于基于G93A小鼠生存的延伸的分子机制知之甚少。我们已经确定了两种化学上不同的（有甲蜂蜜）化合物，它们在疾病发作时显着扩展了G93A小鼠的存活（类似于开始治疗的何时开始）。根据我们的结果，正在努力在具有这些化合物的ALS中进行人类临床试验。我们建议使用这些高效的化合物来探测保护的机制，同时继续我们对SOD1突变体介导的毒性机制的工作。我们将检查蛋白质表达变化，这些变化伴随着这些化合物的保护以及最终的保护损失（AIM 1），以便更好地了解所涉及的机制。这些化合物的寿命延伸幅度可与任何化合物，即使是被持有的那些均可竞争。但是，可靠的比较需要可比的治疗方案。我们建议测试在发作给药范式中赋予有效性的化合物（AIM 2），以测试某些人类试验失败可能与研究设计差异有关的假设。令人信服的新证据表明，线粒体对突变体的摄取对疾病至关重要，并且摄取由金属状态（主要是锌）和内部二硫化物的状态确定。在本文中，我们提供了从根本上提供的新证据，表明体内锌的突变体与疾病的严重程度直接相关，而突变体SOD1（而不是野生型）则是共价交叉的。 AIM 3将识别交叉连接的蛋白质，并确定药物治疗是否改变了它。 AIM 4将使用从脊髓纯化为“体内表达系统”的酶扩展我们对体内金属状态和突变体的硫醇状态的研究。