Extracellular alpha-synuclein in Parkinson's Disease

描述(由申请人提供)：本申请的目标是研究细胞外α-突触核蛋白对帕金森病(PD)神经退行性变和疾病进展的影响。靶向细胞外α-突触核蛋白用于PD新型治疗药物的开发有几个原因，包括在PD患者移植后几年的健康移植神经元中发现含有α-突触核蛋白的路易小体，这表明α-突触核蛋白的传递可能在疾病的传播和进展中发挥作用。我们提出了互补的方法，不仅要阐明α-突触核蛋白释放的基本机制和种类，而且要开发针对细胞外α-突触核蛋白寡聚体的独特治疗干预措施，以期了解细胞外α-突触核蛋白生物学可以导致新的或不同的治疗方法的翻译。我们将检查三个主要目标：第一个目标将使用一系列生物物理和分子工具来定义从神经元释放的α-突触核蛋白的类型，这些工具允许表征释放的α-突触核蛋白的寡聚体形式。我们还将研究α-突触核蛋白释放和摄取的机制以及细胞外α-突触核蛋白对细胞活力的影响。第二个目标是对我们的初步研究和最近发表的研究进行扩展，以探索参与α-突触核蛋白释放和摄取的途径，特别是宏观自噬在α-突触核蛋白释放和摄取中的作用。最后，第三个目标利用体内微透析和AAV引入的在体外具有神经毒性的α-突触核蛋白形式，在体内扩展了这些观察结果，并进一步扩展了宏观自噬可以调节体内α-突触核蛋白释放和摄取的假设。我们将能够共同确定哪些种类的α-突触核蛋白从细胞中释放并最终被邻近细胞摄取；与释放和摄取相关的机制；以及影响α-突触核蛋白释放和摄取的操作是否会影响α-突触核蛋白在细胞和动物模型中诱导的毒性，目标是转化为临床。

DESCRIPTION (provided by applicant): The goal of this application is to investigate the impact of extracellular alpha-synuclein on neurodegeneration and disease progression in Parkinson's disease (PD). There are several reasons to target extracellular alpha-synuclein for the development of novel therapeutic agents for PD including the fact that Lewy bodies containing alpha-synuclein have been found in healthy grafted neurons in PD patients several years after transplant suggesting a possible role for alpha-synuclein transmission in disease propagation and progression. We have proposed complementary approaches to elucidate not only the fundamental mechanisms and species involved in alpha-synuclein release, but to develop unique therapeutic interventions that target extracellular alpha- synuclein oligomers with a view that understanding extracellular alpha-synuclein biology can lead to a translation of new or different therapeutic approaches. We will examine 3 major aims: The first aim will define the types of alpha-synuclein released from neurons using a series of biophysical and molecular tools that allow characterization of released oligomeric forms of alpha-synuclein. We will also investigate the mechanisms involved in the release and uptake of alpha-synuclein and the impact of extracellular alpha-synuclein on cell viability .The second aim will expand on our preliminary studies and on recently published studies to investigate pathways involved in alpha-synuclein release and uptake and in particular the role of macroautophagy in alpha-synuclein release and uptake. Finally, the third aim extends these observations in vivo, utilizing in vivo microdialysis and AAV introduced forms of alpha-synuclein that are neurotoxic in vitro, and furthermore extends the hypothesis that macroautophagy can modulate alpha-synuclein release and uptake in vivo. Together, we will be able to determine which species of alpha-synuclein are released from cells and ultimately taken up by neighboring cells; the mechanisms associated with release and uptake; and if manipulations affecting alpha-synuclein release and uptake affect alpha-synuclein-induced toxicity in cells and animal models with the goal of translation into the clinic.