Enhancing diagnosis of Parkinson's disease and multiple system atrophy via detection of alpha-synuclein seeding activity

突触核蛋白病如帕金森病（PD）和多系统萎缩症（MSA）涉及异常的神经元凋亡。 蛋白质α-突触核蛋白（α-syn）的积累。尽管PD已经有近两个世纪的历史了， 首先描述的是，生物标志物的缺乏极大地阻碍了疾病修饰的发展。 正确的治疗和早期诊断。这一建议是基于这样的假设，即这些疾病中的病理学 疾病通过一种机制在大脑中传播，在这种机制中，错误折叠的α-syn 出现蛋白质聚集体或“种子”。也有证据表明，致病性α-syn具有不同的结构， 疾病，如PD或MSA，这可能是这些疾病中观察到的临床和病理差异的基础。 共核蛋白病该提案利用了一种新的基于细胞的检测方法，该方法已被证明可以检测和定量 在患者脑提取物中，生物化学和结构上均不同的异常α-syn播种活性 警局和海事局的人在这个项目中，我们建议（1）优化检测α-syn聚集形式的能力 利用基于细胞的传感器的遗传操作，（2）利用尖端分子和成像技术 分析突触核蛋白病MSA和PD中α-syn之间的差异，以及（3）确定α-syn是否 患者样品的血液成分中的接种活性可以作为临床疾病进展的标志。 这项工作的意义包括促进PD和MSA患者的诊断和治疗。 申请人是肯塔基州大学神经病学系的助理教授， 致力于作为神经退行性疾病领域的独立翻译科学家的学术生涯 疾病拟议的培训将利用退伍军人事务部丰富的协作环境， 优秀的临床文件和记录系统，以及受这些影响的非常相关的患者人群 疾病，并在VA的临床随访：所有这些都是成功的关键因素， 提议培训将在经验丰富的导师团队的指导下进行。约翰·斯莱文博士， 主要导师，并拥有超过40年的运动障碍临床研究的专业知识，并持有 在基础科学研究方面获得退伍军人管理局的优异奖。西德尼·怀特哈特博士和朱海宁博士将担任联合... 导师他们分别是血小板生化和异常蛋白聚集的专家，持有功勋 在各自领域的奖项，也是大学分子和细胞生物化学系的教师 来自肯塔基州。这些导师是专门挑选出来的，以提高候选人的分子生物学知识。 研究运动障碍患者外周液中聚集蛋白的技术。 在此期间的建议培训包括先进的遗传，分子和成像技术的蛋白质 扩增和检测的技术，以及用于分离和分析外来体和血小板级分的技术， 全血。这些组成部分将是必不可少的，以完成申请人的近期目标，发展一个 可以通过利用生物标志物检测和区分外周样品中的突触核蛋白病 α-syn播种活性的差异。全面的培训计划将使申请人能够进行 独立研究PD和相关神经退行性疾病的新型生物标志物。

Synucleinopathies such as Parkinson's disease (PD) and multiple systems atrophy (MSA) involve abnormal accumulation of the protein alpha-synuclein (α-syn). Although it has been nearly two centuries since PD was first described, a dearth of biomarkers has greatly impeded progress toward development of disease-modifying treatments and accurate and early diagnosis. This proposal is based on the hypothesis that pathology in these diseases propagates through the brain via a mechanism in which transcellular movement of misfolded α-syn protein aggregates or “seeds” occurs. There is also evidence that pathogenic α-syn has diverse structures in diseases such as PD or MSA, that may underlie the clinical and pathologic differences seen in these synucleinopathies. This proposal utilizes a novel cell-based assay that has been shown to detect and quantify abnormal α-syn seeding activity that differs both biochemically and structurally in brain extracts from patients with PD and MSA. In this project we propose to (1) optimize the ability to detect aggregated forms of α-syn utilizing genetic manipulation of a cell-based sensor, (2) utilize cutting edge molecular and imaging techniques to parse out differences between α-syn in the synucleinopathies MSA and PD, and (3) determine whether α-syn seeding activity in blood components of patient samples can serve as a marker for clinical disease progression. The implications of this work include facilitation of diagnosis and therapeutics for patients with PD and MSA. The applicant is an Assistant Professor in the Department of Neurology at University of Kentucky, who is firmly committed to an academic career as an independent translational scientist in the area of neurodegenerative disease. The proposed training will take advantage of the rich and collaborative environment at the VA, the excellent clinical documentation and records system, and the very relevant patient population affected by these diseases and followed in clinic at the VA: all are factors that are integral and crucial to the success of this proposal. Training will be conducted under the direction of an experienced mentor team. Dr. John Slevin, will be the primary mentor, and has over 40 years of expertise in clinical research in Movement Disorders and holds a Merit Award at the VA in basic science research. Dr. Sidney Whiteheart and Dr. Haining Zhu will serve as co- mentors. They are experts in platelet biochemistry and abnormal protein aggregation, respectively, hold Merit Awards in their fields, and are also faculty in the Department of Molecular and Cellular Biochemistry at University of Kentucky. These mentors have been specially selected to advance the candidate’s knowledge of molecular techniques for investigating aggregated proteins in peripheral fluids of patients with movement disorders. Training during this proposal includes advanced genetic, molecular, and imaging-based techniques for protein amplification and detection, and techniques for isolation and analysis of exosomal and platelet fractions from whole blood. These components will be essential to completing the applicant’s immediate goal of developing a biomarker that can detect and differentiate between synucleinopathies in peripheral samples by utilizing differences in α-syn seeding activity. The comprehensive training program will position the applicant to conduct independent research in the study of novel biomarkers in PD and related neurodegenerative disease.