Patient Motor Neuron Assay System for ALS Drug Discovery

项目总结/摘要 肌萎缩侧索硬化症（ALS），也称为卢伽雷病，是一种毁灭性的神经系统疾病， 患者通常在诊断后仅存活三到五年。约3万 美国人患有ALS，每年另有5，600人被诊断出（ALS协会）。尽管身份 在过去的动物模型和永生化人类细胞系研究中， 二十年来，只有一种FDA批准的药物利鲁唑（利鲁唑）可用于治疗这种疾病， 患者的生存期可延长两到三个月。这种令人清醒的状况凸显了对小说的迫切需求 药物疗法，也许通过一个新的药物发现平台。 BrainXell Inc.旨在建立一个自然的体外筛选平台，以确定有前途的ALS药物 引导进一步发展。通过从ALS患者中产生诱导多能干细胞（iPSC），我们 发现iPSC衍生的运动神经元（MN）表现出神经丝（NF）聚集， 轴突变性和最后的MN死亡，使人想起ALS患者中所见的主要病理学。我们 进一步发现NF的一个亚基NF轻链（NF-L）降低，如果NF-L水平恢复，NF 即使在疾病突变存在的情况下，聚集和神经突变性也得到减轻。因此，NF 调节不当是NF聚集和MN变性的关键原因，因此是一种潜在的治疗方法。 目标该公司最近用报道基因纳米荧光素酶（nanoluciferase，100 比传统的荧光素酶灵敏10倍），与NF-L融合以允许简单、稳健地读出NF-L 蛋白质水平它最近开发了一种技术，可以产生大量的运动神经元（数十亿美元）。 一批），使高通量筛选（HTS）成为可能。这项建议将建立在 这些技术，并确定建立一个HTS平台的可行性ALS药物发现使用 患者MN。这项工作可能代表了有史以来第一个基于患者细胞的HTS平台。它将提供一个 有机会在生物系统中筛选化合物库，因为它更接近疾病， 存在于人。

Project Summary/Abstract Amyotrophic lateral sclerosis (ALS), also known as Lou Gehrig’s disease, is a devastating neurological disorder with patients typically surviving just three to five years after diagnosis. Approximately 30,000 Americans live with ALS and another 5,600 are diagnosed each year (ALS Association). Despite identification of over 70 candidate drugs from studies with model animals and immortalized human cell lines over the past two decades, only a single FDA-approved drug, riluzole (Rilutek), is available to treat the disease, increasing survival of patients by two to three months. This sobering status highlights an urgent need of novel pharmaceutical therapies, perhaps via a new drug discovery platform. BrainXell Inc. seeks to create a naturalistic in vitro screening platform to identify promising ALS drug leads for further development. By generating induced pluripotent stem cells (iPSCs) from ALS patients, we discovered that the iPSC-derived motor neurons (MN) exhibit neurofilament (NF) aggregation followed by axonal degeneration and finally MN death, reminiscent of the cardinal pathology seen in ALS patients. We further found that a subunit of NF, NF light chain (NF-L) was decreased, and if NF-L level was restored, NF aggregation and neurite degeneration were mitigated, even in the presence of disease mutations. Thus, NF misregulation is a critical cause of NF aggregation and MN degeneration, and hence a potential therapeutic target. The company has recently engineered ALS patient iPSCs with a reporter gene, nanoluciferase (100 times more sensitive than the traditional luciferase), fused to NF-L to allow for a simple, robust readout of NF-L protein level. It has recently developed a technology to generate large quantities of motor neurons (billions in one batch) from patient iPSCs, making high-throughput screening (HTS) possible. This proposal will build upon these technologies and determine the feasibility of establishing an HTS platform for ALS drug discovery using patient MNs. This effort likely represents the first ever patient cell-based HTS platform. It will provide an opportunity to screen compound libraries in a biological system that more closely resembles the disease as it exists in man.