Diversity Supplement to R01NS105774

摘要 α-突触核蛋白的表达和神经元谷胱甘肽水平都被认为是至关重要的 帕金森病(PD)发病机制中的因素，但这些因素是如何相互作用的 人们对此知之甚少。这一知识差距具有治疗意义，因为 恢复神经元谷胱甘肽水平的药理药物已经在临床上可用。在……里面 PD，α-突触核蛋白寡聚体或高阶聚集体的聚集与 氧化应激，谷胱甘肽耗竭，神经元死亡。此外，通关 α-突触核蛋白的活性受到酪氨酸激酶c-Abl的破坏，该蛋白由氧化激活 压力。谷胱甘肽是主要的硫醇氧化还原物种，被细胞用来 清除活性氧，修复氧化损伤的蛋白质。在这里我们的目标是 确定神经元硫醇氧化还原状态、c-Abl之间的特定因果关系 激活和α-突触核蛋白介导的病理学。我们的基本假设是α- 突触核蛋白聚集体通过金属催化形成活性氧 过程中，所产生的谷胱甘肽耗竭有助于α-突触核蛋白在 前馈的方式。这项研究将使用细胞培养模型，在该模型中，α-突触核蛋白 表达和硫醇氧化还原状态是可以控制和监测的。这些研究还将使用 THY-1α-突触核蛋白“61系”杂交产生的新型双转基因小鼠 小鼠，表现出α-突触核蛋白聚集体形成，与EAAT3-/-小鼠， 表现出较低的神经元谷胱甘肽水平。这些小鼠的谷胱甘肽缺陷可能是 用口服半胱氨酸前体药物逆转。这允许在实验中对 神经元原位硫醇氧化还原状态和潜在疾病修饰的评估 帕金森病的治疗剂。 1

ABSTRACT Both α-synuclein expression and neuronal glutathione levels are recognized as crucial factors in the pathogenesis of Parkinson’s disease (PD), but how these factors interact is poorly understood. This knowledge gap is of therapeutic relevance, because pharmacological agents for restoring neuronal glutathione levels are clinically available. In PD, the accumulation of α-synuclein oligomers or higher-order aggregates is associated with oxidative stress, glutathione depletion, and neuronal death. Additionally, the clearance of α-synuclein is impaired by the tyrosine kinase c-Abl, which is activated by oxidative stress. Glutathione is the dominant thiol redox species and is used by cells to both scavenge reactive oxygen species and repair oxidatively damage proteins. Here we aim to identify specific cause-effect relationships between neuronal thiol redox status, c-Abl activation, and α-synuclein - mediated pathology. Our underlying hypothesis is that α- synuclein aggregates drive reactive oxygen species formation through metal-catalyzed processes, and the resulting glutathione depletion contributes to α-synuclein aggregation in a feed-forward manner. The studies will use cell culture models in which both α- synuclein expression and thiol redox state can be controlled and monitored. The studies will also use a novel double-transgenic mouse generated by crossing the Thy-1 α-synuclein “Line 61” mouse, which exhibits α-synuclein aggregate formation, with the EAAT3-/- mouse, which exhibits low neuronal glutathione levels. The glutathione defect in these mice can be reversed with oral cysteine pro-drugs. This permits both experimental manipulation of neuronal thiol redox state in situ, and an assessment of potential disease -modifying therapeutic agents for PD. 1