Pathological Changes of Alpha-Synuclein Structure in the Brain

大脑α-突触核蛋白结构的病理变化

• 批准号: 9788107
• 负责人: DENNIS J SELKOE
• 金额: $ 22.38万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-09-30 至 2020-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9788107
• 关键词: Affect; Amyloid; Biochemical; Biological Assay; Biological Markers; Brain; Brain Diseases; Cell Culture Techniques; Cell model; Cells; Cerebellum; Characteristics; Collaborations; Communities; Complement; Complex; Corpus striatum structure; Coupled; Cytosol; Data; Detection; Diagnosis; Diffuse; Discipline; Disease; Drug Design; Electron Microscopy; Ensure; Environment; Fingerprint; Genetic; Gold; Human; In Vitro; Inherited; Knowledge; Letters; Lewy Bodies; Lewy Body Dementia; Link; Lip structure; Locales; Mass Spectrum Analysis; Measures; Membrane; Methods; Modeling; Molecular; Molecular Weight; Multiple System Atrophy; Neurites; Neurons; Parkinson Disease; Parkinson' s Dementia; Pathogenicity; Pathologic; Pathology; Patients; Physiological; Point Mutation; Positioning Attribute; Prions; Procedures; Process; Production; Proteins; Proteolysis; Reagent; Recombinants; Research; Resolution; Sampling; Seeds; Source; Specificity; Spectrum Analysis; Structure; Structure-Activity Relationship; System; Testing; Therapeutic; Time; Tissue Extracts; Tissues; Toxic effect; Toxicity Tests; alpha synuclein; alpha synuclein gene; amyloid formation; base; biological systems; biophysical properties; cerebral atrophy; disorder subtype; early onset; human disease; in vivo; in vivo Model; neuropathology; novel; prion-like; protein aggregate; protein misfolding cyclic amplification; screening; structural biology; synucleinopathy

! Aggregates of alpha-synuclein (αSyn) are a pathological hallmark of Parkinson's disease (PD), Dementia with Lewy Bodies (DLB) and Multiple System Atrophy (MSA). Moreover, genetic increases in αSyn expression and point mutations in the αSyn gene are an increasingly well-documented precipitant of early- onset familial PD (fPD). Pathological oligomers have been extensively studied, but only detailed analyses of synthetic aggregates from recombinant sources are available. Therefore, we wish to characterize pathological αSyn species we are isolating from PD/DLB and MSA brains. We plan here an extensive characterization of toxic and self-permissive aggregated forms of αSyn, isolated from human patient brain. Characterization of the isolated species will be both functional (toxicity) and structural for a correlative analysis of structure-function relationship. Additional analysis of different brains will allow a comparison between structure specificity of the different diseases (disease “strains”). The isolation procedure will focus on gentle, non-denaturing methods targeting 3 different sub-cellular locales (cytosol, membrane- associated, insoluble). Results from our novel concept could point to exciting possibilities in rational drug design based on exact structural details of pathogenic strains. Detection and quantification the pathogenic αSyn oligomers may be used as a biomarker for diagnosis of synucleinopathies and will provide novel reagents to the community along with valuable platforms (strain specific PMCA assays) for therapeutic compound screening. To move our hypotheses forward, we propose to gather data in two major directions: Aim 1: Search for the existence of natively soluble `pathological seeds' of αSyn in human brain homogenates of PD, DLB and MSA patients under non-denaturing conditions and quantify their bioactivity in dynamic assays of pathogenic aggregation and toxicity. Aim 2: Characterize the unique structural features (“strains”) of the brain isolated insoluble aggregates from each disease subgroup (PD/DLB/MSA) and their amplification products to contrast their biochemical and biophysical properties in relation to disease. Results from our novel concept could point to exciting possibilities in rational PD drug design based on exact structural details of pathogenic strains. In addition, detection and quantification the pathogenic αSyn oligomers may be used as a biomarker for diagnosis of synucleinopathies and will provide novel reagents (synthetic human brain derived strains) to the community along with valuable platforms (strain specific detection assays) for therapeutic compound screening. ! !

! α-突触核蛋白（αSyn）的聚集体是帕金森病（PD）、痴呆（Dementia 路易体（DLB）和多系统萎缩（MSA）。此外，αSyn的遗传增加 αSyn基因的表达和点突变是越来越多的证据证明的早期- 发病家族性PD（FPD）。病理性寡聚体已被广泛研究，但仅详细分析 来自重组来源的合成聚集体是可获得的。因此，我们希望描述 我们从PD/DLB和MSA脑中分离的病理性αSyn种类。我们在这里计划一个广泛的 从人类患者脑中分离的αSyn的毒性和自我允许聚集形式的表征。 分离物种的表征将是功能（毒性）和结构的相关性 结构-功能关系分析。对不同大脑的额外分析将允许进行比较 不同疾病（疾病“株系”）的结构特异性之间的关系。隔离程序将集中在 在温和的，非变性的方法，靶向3个不同的亚细胞区域（细胞质，膜， 相关的，不溶的）。我们的新概念的结果可以指出合理药物的令人兴奋的可能性 设计基于致病菌株的确切结构细节。检测和量化病原体 αSyn寡聚体可用作诊断突触核蛋白病的生物标志物，并将提供新的 沿着有价值的平台（菌株特异性PMCA测定）， 化合物筛选为了推进我们的假设，我们建议在两个主要方面收集数据。 方向：目的1：寻找αSyn在人脑中天然可溶的“病理种子”的存在 在非变性条件下测定PD、DLB和MSA患者的匀浆并定量其生物活性 在病原体聚集和毒性的动态测定中。目标2：描述独特的结构 从每个疾病亚组（PD/DLB/MSA）分离的脑不溶性聚集体的特征（“菌株”） 以及它们的扩增产物，以对比它们的生物化学和生物物理性质， 疾病从我们的新概念的结果可以指出在合理的PD药物设计令人兴奋的可能性 基于致病菌株的精确结构细节。此外，检测和定量 致病性αSyn寡聚体可用作诊断突触核蛋白病的生物标志物， 新型试剂（合成人脑衍生菌株）沿着有价值的平台 （菌株特异性检测测定）用于治疗性化合物筛选。 ! !