Extracellular alpha-synuclein in Parkinson's Disease

描述（由申请人提供）：本申请的目的是研究细胞外α -突触核蛋白对帕金森病（PD）神经变性和疾病进展的影响。以细胞外α -突触核蛋白为靶点开发新的PD治疗剂有几个原因，包括在PD患者移植后数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起作用。我们提出了互补的方法，不仅阐明了参与α -突触核蛋白释放的基本机制和物种，而且开发了针对细胞外α -突触核蛋白寡聚物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。我们将研究3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α -突触核蛋白释放的低聚形式。我们还将研究α -突触核蛋白的释放和摄取机制以及细胞外α -突触核蛋白对细胞活力的影响。第二个目标将扩展我们的初步研究和最近发表的研究，以调查α -突触核蛋白释放和摄取的途径，特别是巨噬在α -突触核蛋白释放和摄取中的作用。最后，第三个目标将这些观察结果扩展到体内，利用体内微透析和AAV引入的α -突触核蛋白在体外具有神经毒性，并进一步扩展了巨噬可以调节α -突触核蛋白在体内释放和摄取的假设。一起，我们将能够确定哪些种类的α -突触核蛋白从细胞中释放出来并最终被邻近细胞吸收；与释放和摄取相关的机制；如果影响α -突触核蛋白释放和摄取的操作影响细胞和动物模型中α -突触核蛋白诱导的毒性，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.