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

帕金森氏病(PD)和多系统萎缩(MSA)等共核病包括异常 蛋白α-突触核蛋白(α-SYN)的积累。虽然距离警局成立已经近两个世纪了 首先描述的是，生物标记物的缺乏极大地阻碍了疾病改良的发展。 治疗和准确的早期诊断。这一建议是基于这样的假设，即这些疾病的病理 疾病通过一种机制通过大脑传播，在这种机制中，错误折叠的α-syn跨细胞运动 蛋白质聚集体或“种子”就会出现。也有证据表明，致病的α-SYN在 PD或MSA等疾病，这些疾病可能是这些疾病临床和病理差异的基础 联体核病。这一建议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量化 患者脑提取液中生化和结构不同的α-SYN种子活性异常 和PD和MSA在一起。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)利用尖端的分子和成像技术 分析α-SYN在共核病MSA和PD中的差异，以及(3)确定α-SYN是否 患者样本血液成分中的种子活性可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。 申请人是肯塔基大学神经病学系的助理教授，他坚定地认为 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和协作的环境， 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群体 疾病和在退伍军人管理局的临床随访：所有这些都是成功的不可或缺的和关键的因素 求婚。培训将在经验丰富的导师团队的指导下进行。约翰·斯莱文博士将成为 主要导师，在运动障碍的临床研究方面拥有40多年的专业知识，并持有 退伍军人事务部基础科学研究优秀奖。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物化学和异常蛋白聚集方面的专家，拥有优点 在各自领域获奖，也是大学分子和细胞生物化学系的教员 肯塔基州。这些导师是被特别挑选出来的，以促进候选人对分子的了解 研究运动障碍患者外周积液中聚集蛋白的技术。 本提案期间的培训包括先进的基于蛋白质的遗传、分子和成像技术 扩增和检测以及分离和分析红豆杉胞外体和血小板组分的技术 全血。这些组件对于完成申请者的近期目标--制定 利用生物标记物检测和鉴别外周样本中的同型核病 α-SYN播种活性的差异。综合培训计划将使申请者能够进行 在帕金森病和相关神经退行性疾病的新生物标志物研究方面的独立研究。

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.