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。靶向细胞外α-突触核蛋白用于开发PD的新型治疗剂有几个原因，包括移植后数年在PD患者的健康移植神经元中发现含有α-突触核蛋白的路易体的事实，这表明α-突触核蛋白传递在疾病传播和进展中的可能作用。我们已经提出了补充的方法，以阐明不仅在α-突触核蛋白释放中涉及的基本机制和物种，而且开发靶向细胞外α-突触核蛋白寡聚体的独特的治疗干预，以期理解细胞外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-突触核蛋白的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。第二个目标将扩展我们的初步研究和最近发表的研究，以研究α-突触核蛋白释放和摄取的途径，特别是在α-突触核蛋白释放和摄取中的作用。最后，第三个目的扩展了这些观察结果在体内，利用体内微透析和AAV引入的形式的α-突触核蛋白是神经毒性的体外，并进一步扩展了假设，即大自噬可以调节α-突触核蛋白的释放和摄取在体内。总之，我们将能够确定哪些种类的α-突触核蛋白从细胞中释放并最终被邻近细胞吸收;与释放和吸收相关的机制;以及影响α-突触核蛋白释放和吸收的操作是否影响细胞和动物模型中的α-突触核蛋白诱导的毒性，目的是转化为临床。

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.