NADPH Oxidase 2 in Parkinson's Disease Pathogenesis

NADPH 氧化酶 2 在帕金森病发病机制中的作用

基本信息

批准号：
10581821
负责人：
Roberto Di Maio
金额：
$ 48.36万
依托单位：
UNIVERSITY OF PITTSBURGH AT PITTSBURGH
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-12-05 至 2027-11-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10581821
关键词：
Address Affect Animal Disease Models Antisense Oligonucleotides Binding Biological Assay Biological Process Brain Cells Complex Consensus Data Defect Detection Disease Disease model Electron Transport Environment Event Experimental Designs Experimental Models Functional disorder Genes Histologic Idiopathic Parkinson Disease In Situ Inflammatory Response Knock-out LRRK2 gene Ligation Mediating Microglia Mitochondria Modeling Movement Disorders NADP NADPH Oxidase Nature Nerve Degeneration Neuroglia Neurons Outcome Oxidases Oxidation-Reduction Oxidative Stress Parkinson Disease Parkinsonian Disorders Pathogenesis Pathogenicity Pathologic Pathology Patients Persons Phosphotransferases Play Post-Translational Protein Processing Printing Process Production Protein Import Protein Isoforms Rattus Reactive Oxygen Species Regulatory Pathway Reporting Research Resolution Role Rotenone Signal Transduction Source Superoxides System Technology Testing Therapeutic Time United States Work age related alpha synuclein design dopaminergic neuron effective therapy knock-down mitochondrial dysfunction neuroinflammation neuroprotection nigrostriatal degeneration non-motor symptom novel novel drug class novel therapeutic intervention overexpression oxidative damage pathogenic isoform pharmacologic prevent response

项目摘要

ABSTRACT Reductive-oxidative (redox) imbalance in nigrostriatal neurons has been indicated as an important cause of nigrostriatal degeneration in familial and idiopathic Parkinson disease (PD). While it is generally assumed that the oxidative damage seen in PD derives from dysfunctional mitochondrial electron transport chain, there is growing consensus that NADPH oxidase isoform 2 (NOX2) may be important and, in some cellular systems, the two appear intimately related in a redox regulatory pathway recently termed “ROS-induced ROS production” (RIRP). However, the relevance of NOX2 in PD pathogenesis remains underexplored. Additionally, the in-situ detection of NOX2 activity state has been difficult to assess. Using a novel histological proximity ligation (PL) assay to detect NOX2 activation state with excellent cellular resolution, our preliminary findings address the relevance of NOX2 activity, in idiopathic PD and in PD animal models of parkinsonism. Our preliminary studies reveal that NOX2 can be activated by mitochondrially-derived ROS in a RIRP manner, which serves to greatly amplify ROS production. The resultant NOX2 activity is necessary and sufficient to both activate wildtype LRRK2 and to cause accumulation and oxidative post-translational modifications of α-synuclein (4-HNE-α-syn and NO-α-syn). The NOX2-activated LRRK2 kinase activity leads to endo-lysosomal and autophagic defects and decreased degradation of toxic species of α-synuclein, such as pSer129-α-synuclein. In this way, NOX2 activity both drives the formation and inhibits the disposal of toxic forms of α-synuclein. In turn, as shown in previous work, these forms of α-synuclein bind to mitochondrial TOM20, blocking protein import and leading to mitochondrial functional alterations and relative increased ROS production. Thus, there appears to be a feedforward cycle, in which NOX2 plays an essential amplification role in PD pathogenesis. However, other NOXs, including isoforms 1 and 4 – also highly expressed in brain - might contribute somehow to this pathogenic process. In this context, the current proposal is designed to further explore and define the exclusive role of NOX2 in PD. Positive outcomes demonstrating the relevance of NOX2 in PD pathogenesis and the efficacy of NOX2-directed therapies using antisense oligonucleotides (ASOs), will reveal potential therapeutic strategies for neuroprotection and ameliorate the PD-related movement disorder. As such, this project has compelling practical significance for PD therapeutics.

抽象的血纹神经元中的还原性氧化（氧化还原）不平衡已被认为是家族性和特发性帕金森氏病（PD）中的纹状体变性。虽然通常认为 PD中看到的氧化损伤来自功能失调的线粒体电子传输链，有越来越多的共识，即NADPH氧化物同工型2（NOX2）可能很重要，并且在某些细胞系统中，在氧化还原调节途径中，两种外观密切相关，最近称为“ ROS诱导的ROS产生” （RIRP）。但是，NOX2在PD发病机理中的相关性仍然没有被逐渐倍增。另外，原位 NOX2活性状态的检测很难评估。使用新型的组织学接近连接（PL）测定以极好的细胞分辨率检测NOX2激活状态，我们的初步发现解决了在特发性PD和PD动物模型中，NOX2活动的相关性。我们的初步研究表明，Nox2可以通过线粒体衍生的ROS激活，以RIRP的方式激活这可以很好地扩大ROS的生产。由此产生的NOX2活动是必要的，并且足以使两者都足够激活野生型LRRK2并引起α-突触核蛋白的累积和氧化后翻译后修饰（4-HNE-α-Syn和NO-α-Syn）。 NOX2激活的LRRK2激酶活性导致内部溶酶体和自噬缺陷和改善α-突触核蛋白有毒物种的降解，例如pser129-α-核蛋白。在这样，NOX2活性既可以驱动形成并抑制α-突触核蛋白的有毒形式的处置。反过来，如先前的工作所示，这些形式的α-突触核蛋白与线粒体TOM20结合，阻断了蛋白的进口和导致线粒体功能改变并相对增加ROS产生。那似乎有馈电循环，其中NOX2在PD发病机理中起着基本的扩增作用。但是，其他NOX，包括同工型1和4（也在大脑中也高度表达）可能会以某种方式贡献对于这个致病过程。在这种情况下，当前的建议旨在进一步探索和定义 NOX2在PD中的独家作用。阳性结果证明了NOX2在PD发病机理中的相关性和NOX2定向的效率使用反义寡核苷酸（ASO）的疗法将揭示潜在的治疗策略神经保护并改善与PD相关的运动障碍。因此，这个项目具有令人信服的实用性 PD治疗的重要性。