Understanding SOD1 Kinetics in Amyotrophic Lateral Sclerosis

了解肌萎缩侧索硬化症中的 SOD1 动力学

基本信息

批准号：
9282516
负责人：
TIMOTHY M. MILLER
金额：
$ 59.59万
依托单位：
WASHINGTON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-06-01 至 2019-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9282516
关键词：
Adult Alzheimer&apos s Disease Amino Acids Amyotrophic Lateral Sclerosis Antisense Oligonucleotides Biological Assay Brain Cerebrospinal Fluid Cessation of life Clinical Clinical Trials Collection DNA Sequence Alteration Data Diagnosis FDA approved Gene Mutation Gene Proteins Gene Targeting General Population Genes Genetic Grant Half-Life Human Inherited International Kinetics Label Mass Spectrum Analysis Measures Methods Modeling Motor Neurons Muscular Atrophy Mutation Mutation Analysis Neurodegenerative Disorders Participant Pathogenesis Pathologic Patients Pharmaceutical Preparations Pharmacodynamics Phase Phase I Clinical Trials Population Proteins Rattus Respiratory Failure Respiratory Muscles Riluzole Spinal Cord Testing Therapeutic Therapeutic Trials Time Work gene therapy human data man mutant novel novel therapeutics patient population pharmacodynamic biomarker phase II trial repository stable isotope superoxide dismutase 1 targeted treatment

项目摘要

Project Summary Amyotrophic lateral sclerosis is an adult onset neurodegenerative disease characterized by loss of motor neurons resulting in stiffness, weakness, muscle atrophy and death from failure of respiratory muscle 3-5 years after diagnosis. The only FDA approved drug for ALS, Riluzole, is only marginally effective. With disappointing therapeutic options, we have developed targeted therapeutic strategies for genetic subsets of ALS, such as those caused by dominantly inherited mutations in the superoxide dismutase 1 gene (SOD1). Our previous data suggest that SOD1 in the cerebral spinal fluid (CSF) will be an excellent pharmacodynamics marker for an SOD1-focused therapeutic approach. One of the main missing pieces in understanding SOD1 as a marker is SOD1 CSF half-life data. The half-life of the protein will aid in clinical trial planning since half-life influences the amount of SOD1 protein reduction and will dictate the best timing of CSF collection for pharmacodynamics measures. We have recently made huge strides in understanding the protein kinetics of SOD1 by establishing a stable isotope amino acid labeling method using mass spectrometry to measure SOD1 half-life in rat models and normal, healthy human controls. While important first of its kind human data, the CSF SOD1 half- life we have established needs to be extended from normal controls to participants with ALS-causing SOD1 mutations. We will determine SOD1 half-life in CSF of 12 participants with SOD1 mutations by analyzing the kinetics of wild type, mutant, and total SOD1 protein. We hypothesize that the mutant half-life will be decreased in participants with SOD1 mutations. Additionally, some pathological studies suggest that SOD1 becomes misfolded in sporadic ALS and is therefore more broadly implicated in ALS. It is imperative to determine whether this is true as the need to develop SOD1-targeted therapeutics would be greatly increased. However, what's needed is an assay to determine SOD1 involvement in living patients. We hypothesize that protein half-life of SOD1 will be such a measure. We will determine SOD1 half-life in CSF of 19 controls and 19 sporadic ALS participants without SOD1 mutations using our kinetic method. If SOD1 half-life is decreased in sporadic ALS compared to controls, this would suggest that these participants may benefit from the SOD1 targeted strategy and we will consider a therapeutic trial in this population as well. With successful completion of this grant, we will have defined SOD1 kinetics in the most important patient population for SOD1 focused clinical trials and tested whether SOD1 half-life is decreased in sporadic ALS.

项目概要肌萎缩侧索硬化症是一种成人发病的神经退行性疾病，其特征是肌萎缩侧索硬化症丧失运动神经元导致僵硬、虚弱、肌肉萎缩和呼吸衰竭死亡诊断后3-5年的肌肉。 FDA 唯一批准的治疗 ALS 的药物利鲁唑 (Riluzole) 的疗效仅微弱有效的。面对令人失望的治疗选择，我们制定了针对性的治疗策略对于 ALS 的遗传亚型，例如由超氧化物显性遗传突变引起的亚型歧化酶 1 基因 (SOD1)。我们之前的数据表明，脑脊液 (CSF) 中的 SOD1 会是针对以 SOD1 为重点的治疗方法的出色药效学标志物。中的一个理解 SOD1 作为标记物的主要缺失部分是 SOD1 CSF 半衰期数据。的半衰期为该蛋白质将有助于临床试验计划，因为半衰期会影响 SOD1 蛋白质的量减少，并将决定收集脑脊液用于药效学测量的最佳时机。我们有最近通过建立稳定的 SOD1 蛋白质动力学模型，在理解 SOD1 的蛋白质动力学方面取得了巨大进展使用质谱同位素氨基酸标记法测量大鼠模型中 SOD1 半衰期以及正常、健康的人类控制。虽然 CSF SOD1 是同类人类数据中的重要数据，但 CSF SOD1 半我们已经建立的生活需要从正常控制扩展到患有 ALS 的参与者 SOD1突变。我们将通过以下方法确定 12 名具有 SOD1 突变的参与者的脑脊液中 SOD1 半衰期：分析野生型、突变型和总 SOD1 蛋白的动力学。我们假设突变体 SOD1 突变的参与者的半衰期会缩短。此外，一些病理学研究表明 SOD1 在散发性 ALS 中发生错误折叠，因此更广泛地涉及肌萎缩侧索硬化症。由于需要开发针对 SOD1 的药物，因此必须确定这是否属实。治疗量将大大增加。然而，我们需要的是测定 SOD1 的方法参与活着的病人。我们假设 SOD1 的蛋白质半衰期就是这样的衡量标准。我们将确定 19 名对照者和 19 名不带 SOD1 的零星 ALS 参与者的脑脊液中 SOD1 半衰期使用我们的动力学方法进行突变。与散发性 ALS 相比，如果 SOD1 半衰期缩短控制，这表明这些参与者可能会从 SOD1 目标策略中受益，并且我们也将考虑在这一人群中进行治疗试验。随着这笔赠款的顺利完成，我们将在以 SOD1 为重点的临床最重要的患者群体中定义 SOD1 动力学试验并测试了散发性 ALS 中 SOD1 半衰期是否缩短。