Molecular Signatures of Amyotrophic Lateral Sclerosis in Skeletal Muscle

描述（由申请人提供）：肌萎缩侧索硬化症（ALS）是一种运动神经元退行性疾病，可导致进行性虚弱和死亡。这种疾病没有特异性的生物标志物，并且由于临床和电生理检查在早期阶段通常不确定，因此诊断通常被延迟。此外，一旦确定诊断，当前用于跟踪患者的工具对于及时检测疾病改善或恶化不敏感。一种可以促进诊断、跟踪疾病进展或两者兼而有之的生物标志物将填补ALS管理中的巨大临床空白，并加快新疗法的临床试验。本申请的长期目标是鉴定分子 肌萎缩侧索硬化症患者肌肉中的特征可以作为疾病生物标志物。在ALS中，在运动神经元死亡和临床发作之前骨骼肌发生变化，例如神经肌肉接头的结构变化、肌肉限制性线粒体功能障碍和纤维萎缩。肌肉是受ALS退行性过程影响的“终末器官”，并且是最容易进行分子研究的。在这里，我们假设肌萎缩侧索硬化症患者肌肉中的基因表达模式包含可以作为该疾病生物标志物的分子特征。这一假设是基于我们使用下一代深度测序对临床特征良好的ALS患者的肌肉样本获得的初步数据。使用非ALS疾病和正常对照样品，我们在ALS样品中鉴定了300多个独特的靶标，包括同种型差异，这将作为本申请的研究基础。我们很好地进行这项研究，因为大量的神经肌肉患者群体，包括ALS和ALS模拟，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点，有以下两个具体目标：具体目标：1。通过对大量ALS和非ALS肌肉样本进行PCR分析，验证通过深度测序鉴定的ALS特异性靶标和亚型差异。第二章：评估ALS患者肌肉组织中经验证靶标的蛋白质表达，并将靶标与G93A SOD1 ALS小鼠肌肉样本相关联。

DESCRIPTION (provided by applicant): 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 established, 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 application 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 application. We are wel 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.