Towards Understanding the Initiating Role of Truncated Alpha-Synuclein in Neurodegeneration

理解截短的 α-突触核蛋白在神经退行性变中的起始作用

• 批准号: 10176335
• 负责人: Zachary A Sorrentino
• 金额: $ 4.02万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-05-16 至 2022-05-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10176335
• 关键词: Affect; Aging; Alzheimer' s Disease; Animal Model; Animals; Appearance; Biochemical; Biological Assay; Cell Culture Techniques; Cell Death; Cell model; Cells; Cerebrum; Characteristics; Data; Dementia; Detection; Disease; Disease Progression; Disease model; Etiology; Gene Transfer Techniques; Goals; Human; Impairment; In Vitro; Inclusion Bodies; Individual; Inflammation; Inherited; Injections; Investigation; Length; Lewy Bodies; Lewy Body Dementia; Light; Modeling; Molecular; Morphology; Mus; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Neurons; Parkinson Disease; Pathogenesis; Pathogenicity; Pathologic; Pathology; Physiological; Post-Translational Protein Processing; Prevention; Process; Property; Proteins; Recombinant adeno-associated virus (rAAV); Reporting; Role; Severities; Tauopathies; Testing; Time; Toxic effect; Transgenic Animals; Viral; Work; alpha synuclein; combat; comparative; economic impact; experimental study; in vivo; mouse model; mutant; new therapeutic target; overexpression; prevent; prion-like; recruit; synucleinopathy; tau Proteins; tau aggregation; therapeutic target

Project Summary/Abstract: Dementia afflicts 47 million individuals worldwide and has an economic impact of more than $800 billion; neurodegenerative diseases for which no disease modifying therapies exist are the most common etiology. α-synuclein (αS) aggregates into toxic fibrils in multiple neurodegenerative diseases where the fibrils form characteristic inclusions implicated in disease progression. Mechanisms through which initial αS fibrils form is unclear, and yet halting their appearance is key to preventing disease. Carboxy-terminal truncation (C-truncation) of αS may be crucial in disease pathogenesis, as it has been repeatedly shown in- vitro that C-truncated αS spontaneously assembles into fibrils more readily than full-length αS. Proteolytic formation of these truncated species, possibly promoted by aging associated lysosomal impairment, may be key to initial pathology formation in disease. Additionally, C-truncated αS is increased in disease suggesting a pathologic role, where 15-20% of αS in inclusions is truncated. Therefore, The Aims of this project are to characterize naturally formed C-truncated species of αS for their ability to aggregate into pathologic fibrils and stimulate synergistic formation of αS and tau pathology in-vivo. Specific Aim one: Investigate the propensity of physiologic C-terminally truncated αS to pathologically aggregate in-vitro and in cellular models of synucleinopathy and tauopathy. Herein, I will characterize the aggregation propensity of naturally formed C-truncated αS both in-vitro and in-vivo using well characterized models. My working hypothesis is that truncated forms of αS will aggregate readily and synergistically induce full-length αS and tau fibril formation. These results will elucidate molecular mechanisms of aggregation and set the stage for further in-vivo work. Specific Aim two: Determine the ability of physiologic C-terminally truncated αS to initiate pathology in mouse models of synucleinopathy and tauopathy utilizing cerebral fibril injection and recombinant adeno associated virus (rAAV). Viral overexpression of familial aggregation prone αS mutants in mice results in neurodegenerative features used to model disease. Additionally, injection of preformed αS fibrils has been found to hasten progression of the disease in transgenic animals. My working hypothesis is that injection of truncated αS fibrils, and conversely rAAV overexpression of truncated αS in mice will initiate αS and tau pathology. These experiments will uncover pathogenic roles for αS truncation in neurodegenerative diseases.

项目摘要/摘要：痴呆症困扰着全球 4700 万人，造成的经济影响为 超过8000亿美元；尚无疾病缓解疗法的神经退行性疾病是 最常见的病因。 α-突触核蛋白 (αS) 在多种神经退行性疾病中聚集成有毒原纤维 其中原纤维形成与疾病进展有关的特征性内含物。通过哪些机制 最初的 αS 原纤维形式尚不清楚，但阻止其出现是预防疾病的关键。羧基末端 αS 的截断（C 截断）可能在疾病发病机制中至关重要，因为它已被反复证明在- 在体外，C-截短的 αS 比全长 αS 更容易自发组装成原纤维。蛋白水解 这些截短物种的形成可能是由衰老相关的溶酶体损伤促进的 疾病初始病理形成的关键。此外，C-截短的 αS 在疾病中增加，表明 病理作用，其中夹杂物中 15-20% 的 αS 被截断。因此，该项目的目标是 表征天然形成的 C 截短的 αS 物种，因为它们能够聚集成病理原纤维，并且 刺激体内 αS 和 tau 病理学的协同形成。 具体目标一：研究生理性 C 末端截短的 αS 到病理性的倾向 聚合突触核蛋白病和 tau 蛋白病的体外和细胞模型。在此，我将描述 使用充分表征的体外和体内自然形成的 C-截短的 αS 的聚集倾向 模型。我的工作假设是，αS 的截短形式很容易聚集并协同诱导 全长 αS 和 tau 原纤维形成。这些结果将阐明聚集的分子机制和 为进一步的体内工作奠定基础。 具体目标二：确定生理性 C 末端截短的 αS 启动病理学的能力 利用脑原纤维注射和重组的突触核蛋白病和 tau 蛋白病小鼠模型 腺相关病毒（rAAV）。小鼠中易于家族聚集的αS突变体的病毒过度表达结果 用于模拟疾病的神经退行性特征。此外，注射预制αS原纤维已 发现加速转基因动物的疾病进展。我的工作假设是注入 截短的 αS 原纤维，相反，rAAV 在小鼠中过表达截短的 αS 将启动 αS 和 tau 病理。这些实验将揭示 αS 截短在神经退行性疾病中的致病作用。