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

基本信息

批准号：
10199914
负责人：
Xiaobo Mao
金额：
$ 13.01万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-15 至 2022-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10199914
关键词：
Acute Affect Affinity Age Aging Amyloid Proteins Atomic Force Microscopy Binding Binding Proteins Biochemical Biochemistry Biological Biophysics Brain Cerebrospinal Fluid Characteristics Chronic Classification Corpus striatum structure Cross-Sectional Studies Data Dementia with Lewy Bodies Diagnosis Digestion Disease Disease Progression Dopamine Electrophysiology (science)Endocytosis Endocytosis Pathway Endopeptidase K Evaluation Functional disorder Funding Gene Activation Growth Human In Vitro Injections Knock-out Knowledge Laboratory Research Lentivirus Lewy Bodies Lipid III Longitudinal Studies Lymphocyte Activation Mediating Methods Microscopy Molecular Structure Morphology Multiple System Atrophy Mus Nature Neurodegenerative Disorders Neurons Operative Surgical Procedures Parkinson Disease Parkinson&apos s Dementia Pathogenesis Pathogenicity Pathologic Pathology Pathway interactions Patients Physiology Property Research Role Sampling Scanning Tunneling Microscopy Site Slice Structure Substantia nigra structure Synapses Synaptic Transmission Tauopathies Techniques Time Toxic effect Training Western Blotting age effect age related age related neurodegeneration alpha synuclein base brain tissue dopaminergic neuron experimental study gain of function in vivo indexing insight lens loss of function mechanical properties molecular imaging nano network dysfunction neuron loss neuronal circuitry nigrostriatal pathway novel strategies overexpression protein misfolding cyclic amplification receptor synucleinopathy tau Proteins transmission process two photon microscopy

项目摘要

“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.

“衰老对致病性α-核蛋白菌株和传播机制的影响”项目摘要：除了α-突触核蛋白的传播机制外，不同α-突触核蛋白的作用令人信服了解衰老对突触核断病发病机理的影响，包括帕金森氏病（PD），痴呆症（PDD），Lewy身体（DLB）和多个系统的PD 萎缩（MSA）对于开发更有效的症状疗法至关重要。在这个K01提案中，三个提出了具体目的以理解衰老的影响：（1）生成和表征来自PD/PDD/DLB/MSA患者的脑组织/CSF的致病菌株特异性α-Syn到对照对照纵向和横截面老化的因素。（2）了解年龄对独特的影响来自PD/PDD/DLB/MSA体外和体内PD/PDD/DLB/MSA的致病性α-SYN菌株。（3）发现变速箱与年龄相关的不同致病性α-突触核蛋白的机制。为了实现3个特定目标，6种方法/步骤需要：（i）Juan Troncoso博士和Liana Rosenthal博士将为候选人提供培训人脑组织/CSF。博士。泰德（Ted）和瓦利纳·道森（Valina Dawson），马克·马特森（Mark Mattson）博士将使用脑组织/脑脊髓液（CSF）中的α-突触核蛋白作为模板，并与蛋白质折叠式环状折叠式折叠率杂在一起扩增（PMCA）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-核蛋白菌株的分子结构，并区分如果可能出现潜在的问题，则差异和替代培训将提供陈王博士。（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博士将提供有关研究纳米机械特性和动态的培训不同α-突触核蛋白菌株的生长特征。（iv）发射实验的电生理研究将是动手关于Antonello Bonci博士的培训。这项培训研究是为了了解内在和突触特性受不同α-核蛋白菌株的影响。（v）体内显微镜将是Da-ting Lin博士的动手训练。该培训包括执行所有必要的外科手术程序，以将梯度指数（Grin）镜头插入黑质和两光子显微镜。这项研究将允许候选人研究多巴胺神经元受到立体定位注射的电路功能障碍在纹状体区域内注射不同的α-突触核蛋白菌株的电路功能障碍评估传输。（vi）淋巴细胞激活基因3（lag3）已被鉴定为α-核蛋白预先形成的原纤维（PFF）受体，因此值得探索lag3是否可以介导不同的α-突触核蛋白菌株。最重要的是，该项目提出的是开发一个独立的研究实验室能够了解错误折叠结构，传输和神经退行性疾病的毒性，并通过成功地开发一项程序性研究线争夺资金。