Project 1

SODl的聚集和原纤化已经涉及肌萎缩侧索硬化的疾病机制。 硬化症（ALS），这是一个主要的新目标，该计划项目更新，以开发更好的生物测定， 研究这些多聚体形式的SOD 1在系统中的毒性，这些系统将与疾病更相关， 人类在项目2中，我们进一步开发和使用人类细胞培养系统， 运动神经元变性的生物学方面？我们最近开发的人类胚胎干细胞衍生的 运动神经元（HESC-MN）系统。这些单元与其他模型系统相比具有明显的优势， ALS中退化的主要细胞类型，它们是完全人类的。细胞表达识别神经元 标记物，表现出成熟运动神经元典型的电生理功能，并且可以与 其他神经元和非神经元细胞。转染这些细胞以表达ALS-SOD 1蛋白，导致 对细胞存活和形态的有害作用。对于这个项目来说，重要的是，我们最近已经证明， 外源添加的ALS-SOD 1蛋白多聚体被很好地吸收。我们将利用这些细胞来研究 在其形成的不同阶段，SOD 1蛋白多聚体和聚集体的毒性，并将其与细胞毒性相关。 运动神经元变性的进展。该研究计划概述了一个高度协作，逐步 评估运动神经元中自发和诱导的突变体和WT SOD 1聚集体形成的方法， 接着研究SOD 1聚集体对神经变性机制的影响， 最后，通过使用SOD 1聚集的药理学抑制剂来研究是否减少了SOD 1 聚集可以防止运动神经元死亡。

Aggregation and fibrillation of SODl have been implicated in disease mechanisms of Amyotrophic Lateral Sclerosis (ALS), and it is a major new goal of this Program Project renewal to develop better biological assays to study the toxicity of these multimeric forms of SODl in systems that will be more relevant to the disease in humans. In Project 2 we further develop and use a human cell culture system that closely models important cell biological aspects of motor neuron degeneration¿our recently developed human embryonic stem cell-derived motor neuron (HESC-MN) system. The cells have distinct advantages over other model systems as they represent the major cell type that degenerates in ALS and they are fully human. The cells express identifying neuronal markers, exhibit electrophysiological function typical for mature motor neurons, and can be co-cultured with other neuronal and non-neuronal cells. Transfection of these cells to express ALS-SODl proteins causes deleterious effeds on cell survival and morphology. Importantly for this project, we have recently shown that exogenously added ALS-SODl protein multimers are taken up quite well. We will utilize these cells to study the toxicity of SODl protein multimers and aggregates at different stages of their formation and relate it to the progression of motor neuron degeneration. This research plan outlines a highly collaborative, step-by-step approach to evaluate spontaneous and induced mutant and WT SODl aggregate formation in motor neurons, followed by an investigation of the consequences of SODl aggregates on neurodegenerative mechanisms and, finally, by using pharmacological inhibitors of SODl aggregation to investigate whether reduced SODl aggregation can prevent motor neuron death.