Diversity Supplement to R01NS105774

R01NS105774 的多样性补充

• 批准号: 10350351
• 负责人: RAYMOND A SWANSON
• 金额: $ 7.3万
• 依托单位: NORTHERN CALIFORNIA INSTITUTE/RES/EDU
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-07 至 2022-06-06
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10350351
• 关键词: ABL1 gene; Antioxidants; Cell Culture Techniques; Cell Death; Cells; Clinical; Cysteine; Defect; Disease; Exhibits; Glutathione; Impairment; In Situ; Knowledge; Mediating; Metals; Modeling; Monitor; Mus; Neurons; Oral; Oxidation-Reduction; Oxidative Stress; Parkinson Disease; Pathogenesis; Pathology; Pharmacology; Process; Prodrugs; Protein Tyrosine Kinase; Proteins; Reactive Oxygen Species; Sulfhydryl Compounds; Therapeutic; Therapeutic Agents; Transgenic Mice; alpha synuclein; cell injury; excitatory amino acid transporter 3; neuron loss; novel; oxidative damage; protein aggregation; repaired

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

抽象的 α-突触核蛋白表达和神经元谷胱甘肽水平被认为是至关重要的 帕金森病 (PD) 发病机制中存在多种因素，但这些因素如何相互作用尚不清楚 不太了解。这种知识差距具有治疗相关性，因为 用于恢复神经元谷胱甘肽水平的药物在临床上是可用的。在 PD，α-突触核蛋白寡聚体或高阶聚集体的积累相关 氧化应激、谷胱甘肽耗竭和神经元死亡。另外，清关 α-突触核蛋白的功能受到酪氨酸激酶 c-Abl 的损害，酪氨酸激酶 c-Abl 被氧化激活 压力。谷胱甘肽是主要的硫醇氧化还原物质，被细胞用来 清除活性氧并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驱动活性氧的形成 过程，由此产生的谷胱甘肽耗尽有助于 α-突触核蛋白聚集 前馈方式。这些研究将使用细胞培养模型，其中 α-突触核蛋白 可以控制和监测表达和硫醇氧化还原状态。研究还将使用 通过与 Thy-1 α-突触核蛋白“61 系”杂交产生的新型双转基因小鼠 小鼠，其表现出 α-突触核蛋白聚集体形成，与 EAAT3-/- 小鼠，其 表现出低神经元谷胱甘肽水平。这些小鼠的谷胱甘肽缺陷可能是 口服半胱氨酸前药可逆转。这允许实验操作 原位神经元硫醇氧化还原状态，以及潜在疾病缓解的评估 PD的治疗剂。 1