Influence of Aging on Pathogenic Alpha-Synuclein Strains and Transmission Mechanism

“衰老对致病性α-突触核蛋白菌株的影响及传播机制”项目摘要： 除了α-突触核蛋白传递机制外，α-突触核蛋白的不同菌株的作用也令人信服， 明白了解衰老对突触核蛋白病发病机制的影响，包括 帕金森病（PD）、PD伴痴呆（PDD）、路易体痴呆（DLB）和多系统 萎缩（MSA）的治疗对于开发更有效的对症治疗至关重要。在K 01提案中，三个 提出了理解老龄化影响的具体目标：（1）产生和表征 PD/PDD/DLB/MSA患者的脑组织/CSF中的致病菌株特异性α-syn与对照相比， 纵向和横截面的老化因素。(2)为了了解年龄对不同的 PD/PDD/DLB/MSA的致病性α-syn菌株的体外和体内研究。(3)为了揭开传输 与年龄相关的不同致病性α-突触核蛋白的机制。为实现3个具体目标，6个方法/步骤 需要：（i）Juan Troncoso博士和Liana Rosenthal博士将为候选人提供处理培训 人脑组织/脑脊液。Ted和Valina Dawson博士以及Mark Mattson博士将使用 以脑组织/脑脊液（CSF）中错误折叠的α-突触核蛋白为模板， 使用PMCA扩增技术，以产生不同的α-突触核蛋白菌株。(ii)扫描隧道显微镜 (STM)将用于成像不同α-突触核蛋白菌株的分子结构，并区分 差异，如果出现潜在的问题，可以从Chen Wang博士那里获得替代培训。 (iii)原子力显微镜（AFM）将用于观察不同α-突触核蛋白的组装形态 菌株Mingdong Dong博士将提供研究纳米机械性能和动态的培训 不同α-synuclein菌株的生长特征。(iv)放电实验的电生理学研究将是手- 安东尼罗·邦奇博士的培训这个训练研究是为了了解内在的和突触的特性 受到不同的α-突触核蛋白株的影响。(v)体内显微镜将由Da-Ting Lin博士进行实践培训。 该培训包括执行所有必要的外科手术，以将梯度折射率（GRIN）透镜插入 黑质和双光子显微镜。这项研究将允许候选人研究多巴胺神经元 立体定向注射不同的α-突触核蛋白株影响纹状体区的回路功能障碍， 评估传输。(vi)淋巴细胞激活基因3（LAG 3）是一种α-突触核蛋白 因此，LAG 3是否能够介导PFF受体的传递是值得探讨的。 不同的α-突触核蛋白株。最重要的是，本项目提出的是建立一个独立的研究实验室 装备，以了解不同菌株的淀粉样蛋白的错误折叠结构，传输和 神经退行性疾病的毒性，并通过成功地开发一个程序化的研究路线， 竞争资金。

“Influence of Aging on Pathogenic α-Synuclein Strains and Transmission Mechanism” Project Summary: Besides α-synuclein transmission mechanism, the role of distinct strains of α-synuclein is compelling to be understood. Understanding the influence of aging on the pathogenesis of synucleinopathies, including Parkinson's disease (PD), PD with dementia (PDD), dementia with Lewy body (DLB) and multiple system atrophy (MSA), is crucial for developing more effective symptomatic therapies. In this K01 proposal, three specific aims are proposed for understanding the influence of aging: (1) To generate and characterize pathogenic strain-specific α-syn from brain tissue/CSF of patients with PD/PDD/DLB/MSA to controls by the factor of aging longitudinally and cross-sectionally. (2) To understand the influence of age on distinct pathogenic α-syn strains from PD/PDD/DLB/MSA in vitro and in vivo. (3) To uncover the transmission mechanism of age-related distinct pathogenic α-synuclein. To achieve the 3 specific aims, 6 methods/steps are required: (i) Dr. Juan Troncoso and Dr. Liana Rosenthal will provide the training to candidate on handling human brain tissue/CSF. Drs. Ted and Valina Dawson, and Dr. Mark Mattson will train the candidate using the misfolded α-synuclein in brain tissue/cerebrospinal fluid (CSF) as templates, and with protein misfolding cyclic amplification (PMCA) technique, to generate distinct α-synuclein strains. (ii) Scanning tunneling microscopy (STM) will be applied for imaging molecular structures of distinct α-synuclein strains and distinguish the differences, and the alternative training will be available from Dr. Chen Wang if the potential problems appear. (iii) Atomic force microscopy (AFM) will be applied for observing assembly morphologies of distinct α-synuclein strains. Dr. Mingdong Dong will provide the training on studying nano-mechanical properties and dynamic growth features of distinct α-synuclein strains. (iv) Electrophysiology study on firing experiment will be hands- on training from Dr. Antonello Bonci. This training study is to understand the intrinsic and synaptic properties affected by distinct α-synuclein strains. (v) In vivo microscopy will be hands-on training from Dr. Da-Ting Lin. This training includes performing all necessary surgical procedure to insert gradient index (GRIN) lens into substantia nigra and two-photon microscopy. This study will allow candidate to study the dopamine neuronal circuit dysfunction affected by stereotaxically injected with distinct α-synuclein strains in striatum region for evaluation the transmission. (vi) Lymphocyte-activation gene 3 (LAG3) has been identified as α-synuclein preformed fibrils (PFF) receptor, so it is worth to explore whether LAG3 can mediate the transmission of distinct α-synuclein strains. Above all, this project proposed is to develop an independent research laboratory equipped to understand the distinct strains of amyloid proteins on misfolded structures, the transmission and the toxicity in neurodegenerative disorders, and to develop a programmatic line of research by successfully competing for funding.