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; 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. ! !

呢 α-核蛋白（αSyn）的聚集体是帕金森氏病（PD），痴呆症的病理标志 Lewy身体（DLB）和多系统萎缩（MSA）。此外，αSyn的遗传增加 αSyn基因中的表达和点突变是越来越有记录的早期精度 发作家族性PD（FPD）。病理性的低聚物已广泛研究，但仅进行详细的分析 可从重组来源提供合成骨料。因此，我们希望描述 病理αSyn物种我们从PD/DLB和MSA大脑中分离出来。我们在这里计划广泛 从人类患者大脑中分离出的αSyn的有毒和自我验证的综合形式的表征。 孤立物种的表征既具有功能（毒性），又是相关性的结构 结构功能关系的分析。对不同大脑的其他分析将允许比较 在不同疾病的结构特异性（疾病“菌株”）之间。隔离程序将集中 在靶向3个不同细胞局部的柔和，非贬值方法上（细胞质，膜 - 相关，不溶性）。我们新颖概念的结果可能指出了理性药物的激动人心的可能性 基于致病菌株的确切结构细节的设计。检测和定量致病性 αSyn低聚物可以用作突触核酸诊断的生物标志物，并将提供新颖的 社区的试剂以及有价值的平台（特定的PMCA分析）进行治疗 复合筛选。为了推动我们的假设向前发展，我们建议将数据收集到两个主要的 方向：目标1：在人脑中寻找αSyn的本地固体“病理种子”的存在 PD，DLB和MSA患者的匀浆，在非降解条件下，并量化其生物活性 在致病性聚集和毒性的动态测定中。目标2：特征独特的结构 来自每个疾病亚组的大脑分离的不溶性骨料的特征（“菌株”）（PD/DLB/MSA） 以及它们的放大产品以对比其生化和生物物理特性 疾病。我们新颖概念的结果可能指出了理性PD药物设计的激动人心的可能性 基于致病菌株的确切结构细节。另外，检测和数量 致病性αSyn低聚物可以用作诊断突触核苷的生物标志物，并将提供 与有价值的平台一起向社区提供的新型试剂（合成人脑衍生菌株） （特异性检测分析）用于治疗化合物筛选。 呢 呢