Molecular Signatures of Amyotrophic Lateral Sclerosis in Skeletal Muscle

肌萎缩侧索硬化症(ALS)是一种运动神经元退行性疾病，不可阻挡地导致 逐渐衰弱和死亡。这种疾病没有特定的生物标志物，诊断通常是 延迟，因为临床和电生理检查在早期往往是不确定的。 此外，一旦建立了诊断，目前用于跟踪患者的工具对于及时 检测到疾病的好转或恶化。一种有助于诊断、跟踪疾病的生物标志物 进展，或两者兼而有之，将填补肌萎缩侧索硬化症治疗方面的一大临床空白，并加快新的临床试验 治疗。这项提议的长期目标是在ALS患者的肌肉中识别 可以作为疾病的生物标志物。在肌萎缩侧索硬化症中，骨骼肌在运动神经元死亡和 临床起病，如神经肌肉连接结构改变，肌肉线粒体受限 功能障碍和纤维萎缩。肌肉是肌萎缩侧索硬化症退变过程中受其影响的“末梢器官” 最容易进行分子研究的地方。在这里，我们假设肌肉中的基因表达模式来自于 ALS患者包含可作为疾病生物标记物的分子特征。这一假设是 根据初步数据，我们使用下一代深度测序技术对临床上的肌肉样本进行了测序 ALS患者的特点很好。使用非肌萎缩侧索硬化症和正常对照样本，我们确定了300多个 肌萎缩侧索硬化症样本中的独特靶点，包括同形方差，将作为研究的基础 求婚。我们处于进行这项研究的有利地位，因为有大量的神经肌肉患者， 包括肌萎缩侧索硬化症和肌萎缩侧索硬化症的模拟物，以及我们对电诊断和肌肉组织学实验室的监督。 在这里，我们将进一步研究我们的分子靶标，具体有以下两个目标： 具体目标： 1.通过执行聚合酶链式反应来验证通过深度测序鉴定的ALS特异性靶点和异构体变异 对大量肌萎缩侧索硬化症和非肌萎缩侧索硬化症肌肉样本的分析。 2：评估肌萎缩侧索硬化症患者肌肉组织中有效靶点的蛋白表达及相关靶点 来自G93A SOD1ALS小鼠的肌肉样本。

Amyotrophic lateral sclerosis (ALS) is a degenerative disease of motor neurons that inexorably leads to progressive weakness and death. There is no specific biomarker for this disease, and the diagnosis is often delayed as clinical and electrophysiological examinations are often inconclusive in the early stages. Furthermore, once a diagnosis is estabished, the current tools to track patients are insensitive for the timely detection of disease improvement or worsening. A biomarker that can facilitate diagnosis, track disease progression, or both, would fill a large clinical gap in ALS management, and expedite clinical trials of novel therapies. The long term goal of this proposal is to identify molecular signatures in muscle of ALS patients that can serve as disease biomarkers. In ALS, changes occur in skeletal muscle prior to motor neuron death and clinical onset, such as structural changes in the neuromuscular junction, muscle restricted mitochondrial dysfunction, and fiber atrophy. Muscle is the "end organ" affected by the degenerative process of ALS and is the most accessible for molecular study. Here we hypothesize that gene expression patterns in muscle from patients with ALS contain molecular signatures that can serve as biomarkers of the disease. This hypothesis is based on preliminary data we obtained using next generation deep sequencing on muscle samples of clinically well characterized ALS patients. Using non-ALS disease- and normal control samples, we identified over 300 unique targets in ALS samples, including isoform variances, that will serve as the basis of study for this proposal. We are well positioned to carry out this study because of the large neuromuscular patient population, including ALS and ALS mimics, and our oversight of the electrodiagnostic and muscle histology laboratories. Here, we will further investigate our molecular targets with the following two specific aims: Specific Aims: 1. To validate ALS-specific targets and isoform variances identified by deep sequencing by performing PCR analysis of a large cohort of ALS and non-ALS muscle samples. 2: To assess protein expression of validated targets in muscle tissues from ALS patients and correlate targets with muscle samples from the G93A SOD1 ALS mouse.