Project I "Mechanisms of Pathological aSyn Transmission"

项目I：“病理性α--突触核蛋白的传播和毒株” 项目负责人：弗吉尼亚·M·Y·李 项目I摘要/摘要 项目I，前身为项目III，重新编号为项目I，以改善我们重新提交的项目的流程 名为“阿尔茨海默病和相关痴呆的α-突触核蛋白品系研究中心”的新U19拨款 宾夕法尼亚大学佩雷尔曼医学院(PSOM)测试传播和菌株 A-突触核病的假说。路易体痴呆(DLB)、帕金森氏病(PD) 痴呆症(PDD)，这里称为路易体(LBD)障碍，均为路易体(LN)和路易体(LN) 神经元内病理由聚集的α-突触核蛋白(ASyn)组成。LBD、PDD与阿尔茨海默病(AD) 伴(AD-aSyn)或不伴LBS(AD-aSyn)是最常见的衰老相关痴呆。此外，LBD， AD aSyn与多系统萎缩(MSA)，以胶质细胞胞浆内包涵体aSyn错误折叠为特征 (GCIS)是主要的神经退行性联核病。这种多样化的临床特征和aSyn 神经退行性疾病患者大脑中的神经病理为该菌株提供了间接支持 一种假说，病理性aSyn采用不同的构象或菌株，可解释临床和 这些疾病之间的病理异质性。在此基础上，我们进一步提出了传播假说。 ASyn菌株来解释LBD、AD aSyn和MSA进展的变异性。然而，这两个假设 需要严格的测试。最近，项目I和项目II的研究人员合作证明了纹状体内 将重组aSyn预制纤维(PFF)注射到野生型(WT)小鼠体内诱导了模板化的 内源性aSyn的错误折叠和聚集，逐渐通过纹状体连接体扩散到 黑质致密部形成PD样LBS和LNS后多巴胺(DA)能神经元丢失 (SNPC)和帕金森病特有的运动障碍。我们的主要观察结果得到了许多其他人的验证 实验室，并扩展到老鼠和非人灵长类动物(见项目中初步的非人灵长类数据 Ii)从而支持传播假说。这些验证性研究还表明，这些小说 模型概括了LBD的发病机制，并将促进LBD的研究。我们还发现了独特的aSyn 人工合成的具有不同构象和生物活性的菌株体外连续繁殖 从重组的aSyn中产生aSyn pff，我们已经分离出不同的人LBD，AD aSyn和MSA 来自LBD和AD的aSyn菌株，以及MSA大脑的遗传和神经病理特征 在项目III和项目IV中进行临床研究，从而合作 链接所有U19项目和核心。这些研究确定了AD、aSyn和LBD大脑中不同的aSyn菌株 指定为aSyn-Lb(LN)，并来自指定为aSyn-GCI的MSA Brain。这些和其他初步的 这些数据为研究新的aSyn毒株传播致病机制提供了框架 U19中心申请。为此，我们将使用生化、生物物理和细胞生物学方法来 确定aSyn-LB和aSyn-GCI菌株的分子特征，并鉴定细胞决定因素 每个菌株的类型特异性，并阐明这些菌株的模板化繁殖机制。

PROJECT I: “Pathological α-Synuclein Transmission and Strains” PROJECT LEADER: VIRGINIA M.-Y. LEE Project I Summary/Abstract Project I, formerly Project III, is renumbered as Project I to improve the flow of the Projects in our re-submitted new U19 grant entitled “Center On Alpha-synuclein Strains In Alzheimer Disease & Related Dementias” at the University of Pennsylvania (Penn) Perelman School of Medicine (PSOM) to test the transmission and strain hypotheses of a-synucleinopathies. Dementia with Lewy bodies (DLB), Parkinson's disease (PD) without and with dementia (PDD), referred to here as Lewy body (LB) disorders (LBD), all share LB and Lewy neurite (LN) intra-neuronal pathology comprised of aggregated α-synuclein (aSyn). LBD, PDD and Alzheimer's disease (AD) with (AD+aSyn) or without LBs (AD-aSyn) are the most common aging related dementias. Moreover, LBD, AD+aSyn and multiple system atrophy (MSA), characterized by misfolded aSyn in glial cytoplasmic inclusions (GCIs), are the major neurodegenerative synucleinopathies. This diversity of clinical features and aSyn neuropathology in brains of neurodegenerative disease patients provides indirect support for the strain hypothesis wherein pathological aSyn adopts different conformations or strains that account for clinical and pathological heterogeneity between these disorders. Further, we propose the transmission hypothesis of aSyn strains to explain the variability in progression of LBD, AD+aSyn and MSA. However, both hypotheses need rigorous testing. Recently Project I and II investigators collaborated to demonstrate that intrastriatal injections of recombinant aSyn preformed fibrils (PFFs) into wildtype (WT) mice induced the templated misfolding and aggregation of endogenous aSyn that spread progressively across the striatal connectome to form PD-like LBs and LNs followed by dopaminergic (DA) neuron loss in the substantia nigra pars compacta (SNpc) and motor impairments characteristic of PD. Our key observations have been validated by many other laboratories and extended to rats and non-human primates (see preliminary non-human primate data in Project II) thereby supporting the transmission hypothesis. These confirmatory studies also showed that these novel models recapitulate LBD pathogenesis and will facilitate LBD research. We also have identified distinct aSyn strains with different conformations and biological activities through the serial in vitro propagation of synthetic aSyn PFFs generated from recombinant aSyn and we have isolated distinct human LBD, AD+aSyn and MSA aSyn strains from LBD and AD+aSyn, as well as MSA brains characterized genetically and neuropathologically by Core C from patients followed by Core B and studied clinically in Projects III and IV thereby collaboratively linking all U19 Projects and Cores. These studies defined distinct aSyn strains from AD+aSyn and LBD brains designated as aSyn-LB (LB+LN) and from MSA brains designated as aSyn-GCI. These and other preliminary data provide the framework for investigating pathogenic mechanisms of aSyn strain transmission in this new U19 Center application. To do this, we will use biochemical, biophysical and cell biological approaches to determine the molecular signatures of aSyn-LB and aSyn-GCI strains as well as identify the determinants of cell type specificities for each strain and elucidate mechanisms for templated propagation of these strains.