Understanding SOD1 Kinetics in Amyotrophic Lateral Sclerosis

项目摘要 肌萎缩性侧索硬化症是一种成人发病的神经退行性疾病，其特征在于缺乏 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。唯一一种FDA批准的治疗ALS的药物，阿舒唑， 有效由于治疗方案令人失望，我们开发了有针对性的治疗策略 对于ALS的遗传子集，例如由超氧化物歧化酶中的显性遗传突变引起的那些， SOD 1基因。我们以前的数据表明，脑脊液（CSF）中的SOD 1将 是一个优秀的药效学标志物的SOD 1为重点的治疗方法。之一 理解SOD 1作为标记物的主要缺失部分是SOD 1 CSF半衰期数据。的半衰期 该蛋白质将有助于临床试验计划，因为半衰期影响SOD 1蛋白质的量 减少，并将决定用于药效学测量的CSF收集的最佳时间。我们有 最近通过建立一个稳定的 同位素氨基酸标记质谱法测定SOD 1在大鼠模型中的半衰期 和正常健康的人类对照。虽然这是第一个重要的人类数据，CSF SOD 1的一半- 我们已经建立的生命需要从正常对照延长到ALS引起的参与者 SOD 1突变。我们将通过以下方法确定12名SOD 1突变受试者CSF中的SOD 1半衰期： 分析野生型、突变体和总SOD 1蛋白的动力学。我们假设突变体 在SOD 1突变的受试者中，半衰期将缩短。此外，一些病理学研究 这表明SOD 1在散发性ALS中发生错误折叠，因此更广泛地涉及 人症必须确定这是否属实，因为需要开发针对SOD 1的 治疗方法将大大增加。然而，我们需要的是一种测定SOD 1的方法， 参与活的病人。我们假设SOD 1的蛋白质半衰期将是这样一个措施。 我们将测定19名对照和19名无SOD 1的散发性ALS参与者的CSF中SOD 1半衰期。 使用我们的动力学方法。如果SOD 1半衰期在散发性ALS中减少， 控制，这表明这些参与者可能受益于SOD 1有针对性的策略， 我们也会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， 我们将在最重要的患者人群中定义SOD 1动力学，用于SOD 1重点临床研究。 试验并测试SOD 1半衰期是否在散发性ALS中减少。

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.