The role of ATP13A2/PARK9 in secretion of exosomes and alpha synuclein

ATP13A2/PARK9 在外泌体和 α 突触核蛋白分泌中的作用

基本信息

批准号：
10261844
负责人：
DIMITRI KRAINC
金额：
$ 64.52万
依托单位：
NORTHWESTERN UNIVERSITY AT CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2016
资助国家：
美国
起止时间：
2016-09-30 至 2021-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10261844
关键词：
ATP phosphohydrolase Adolescent Affect Amygdaloid structure Anterior Astrocytes Behavioral Binding Proteins Biogenesis Cell Fractionation Cell physiology Cells Coculture Techniques Collaborations Confocal Microscopy Data Degradation Pathway Dementia Disease Early Endosome Endosomes Enzymes Exocytosis Fingers Functional disorder Generations Homeostasis Human Hydrolase Immunohistochemistry Impairment Individual Knockout Mice Levodopa Lysosomes Mediating Medical Genetics Membrane Modeling Mus Nerve Degeneration Neurodegenerative Disorders Neuronal Dysfunction Neurons PARK9 gene Parkinson Disease Parkinsonian Disorders Pathogenesis Pathologic Pathway interactions Patients Phenotype Phosphatidylinositols Physiological Play Proteins Recombinants Research Project Grants Role Syndrome System Testing Transgenic Mice Validation Vesicle Yeasts Zinc alpha synuclein cofactor dopaminergic neuron endosome membrane entorhinal cortex exosome experimental study gaze palsy improved in vivo induced pluripotent stem cell insight loss of function loss of function mutation lysosomal proteins monomer mouse model mutant olfactory bulb olfactory nuclei overexpression pre-clinical protein aggregation protein degradation protein transport recruit synucleinopathy therapeutic target transmission process uptake

项目摘要

Neurodegenerative disorders are characterized by the accumulation of misfolded aggregated proteins in neurons. Since neurons are permanently postmitotic, efficient intracellular protein degradation systems are critically important for normal neuronal function. Recent evidence suggests that disruption of lysosomal degradation pathways directly contributes to neurodegeneration in Parkinson's disease and related synucleinopathies. We have previously shown that loss of function of lysosomal ATPase PARK9 (ATP13A2) leads to zinc dyshomeostasis, lysosomal dysfunction and a-syn accumulation. In addition, we and other found that PARK9 localizes to multivesicular endosomes and regulates exosome biogenesis. Here, we propose to further analyze the physiological role of PARK9 in generation and secretion of exosomes and how loss of PARK9 function contributes to neuronal dysfunction and neurodegeneration. First, we will test the hypothesis that PARK9 plays an important role in the formation of intraluminal vesicles by recruitment of zinc-dependent FYVE proteins to early endosomes. Second, we will examine if a-syn secretion via exosomes and lysosomal exocytosis contributes to PARK9-mediated neuronal dysfunction. Finally, we will test the hypothesis that PARK9 is protective in synucleinopathies by overexpressing PARK9 in mouse models that accumulate a-synuclein. We will also examine propagation of a-synuclein in PARK9 knockout and transgenic mice. These findings will also provide further mechanistic insights into PARK9 loss of function in the context of Kufor-Rakeb syndrome as well as more general forms of syncleinopathies such as Parkinson's disease (PD), especially in terms of cell- to-cell transmission of a-syn that has been implicated in the pathogenesis of these disorders.

神经退行性疾病的特征是错误折叠的聚集体的积累神经元中的蛋白质。由于神经元永久处于有丝分裂后，有效的细胞内蛋白质降解系统对于正常神经元功能至关重要。最近的证据表明溶酶体降解途径的破坏直接导致帕金森病和相关突触核蛋白病中的神经变性。我们之前有过研究表明，溶酶体 ATP 酶 PARK9 (ATP13A2) 功能丧失会导致锌体内平衡失调、溶酶体功能障碍和 a-syn 积累。此外，我们和其他发现 PARK9 定位于多囊泡内体并调节外泌体生物发生。在此，我们建议进一步分析PARK9在生成和分泌中的生理作用外泌体的作用以及 PARK9 功能的丧失如何导致神经元功能障碍和神经变性。首先，我们将检验 PARK9 在通过招募锌依赖性 FYVE 蛋白早期形成管腔内囊泡内体。其次，我们将检查 a-syn 是否通过外泌体和溶酶体分泌胞吐作用导致 PARK9 介导的神经元功能障碍。最后，我们将测试假设 PARK9 通过在小鼠中过度表达 PARK9 而对突触核蛋白病具有保护作用积累α-突触核蛋白的模型。我们还将检查α-突触核蛋白在 PARK9 敲除和转基因小鼠。这些发现也将提供进一步的机制对 Kufor-Rakeb 综合征等情况下 PARK9 功能丧失的见解肌细胞病的一般形式，例如帕金森病 (PD)，特别是在细胞- a-syn 向细胞的传递与这些疾病的发病机制有关。