Mitochrondrial Etiologies of Parkinsons Disease

帕金森病的线粒体病因

• 批准号: 6905373
• 负责人: GEORGE F WOOTEN
• 金额: $ 126.74万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-09-30 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6905373
• 关键词: Parkinson' s disease; clinical research; disease /disorder model; hybrid cells; mitochondrial disease /disorder; molecular pathology

DESCRIPTION (provided by applicant): The major hypothesis examined by the University of Virginia Udall Parkinson's Center is that mitochondrial genome mutations contribute substantially to the pathogenesis of sporadically occurring Parkinson's disease (PD), which is the form afflicting >95% of PD patients. During the first funding period, substantial progress was made in demonstrating with platelet mitochondrial DNA (mtDNA)-derived cybrids that expression of PD mitochondrial genomes reproduces multiple essential pathobiological features of PD. Most dramatically, PD cybrids spontaneously generate Lewy inclusion bodies that are indistinguishable biochemically and morphologically from those present in PD brain. Difficulties in acceptance of the mitochondrial genomic contribution to PD derive in part from the lack of specific mtDNA mutations yet defined in PD brain, concerns about the relevance of PD platelet mtDNA to events in brain neurons, inability to provide direct proof of causality of mtDNA mutations and lack of understanding of how bioenergetic deficits and proteasomal dysfunction can arise in PD from mtDNA mutations. All of these concerns are systematically and comprehensively addressed in this renewal application. Dr. Bennett's project utilizes novel technologies created at UVA that allow the insertion into mitochondria and expression in cells of human mitochondrial genomes from any source including postmortem brain. Cell lines expressing mtDNA from well characterized PD brains will be created and analyzed phenotypically, allowing a direct test of causality. Mutations can also be created by mutagenesis and expressed in cells, directly demonstrating pathogenecity of mtDNA. Dr. Capaldi's project involves a collaboration with the University of Oregon to provide a detailed proteomic analysis of complex I macromolecular assembly and oxidative damage in PD brain mitochondria compared to mitochondria from cells that express mtDNA from the same PD brains. Project 3 will uncover mechanisms of bioenergetic complex I impairment arising from PD mtDNA mutations. Dr. Trimmer's project examines the appearance of proteasomal impairment in individual cells of PD cybrids and cell lines created from PD brain, and defines how proteasomal impairment relates to development of pathogenic protein aggregates. This project specifically links PD mtDNA expression to neuronal PD pathobiology. All projects are supported by two Cores. The Cell Culture Core provides centralized culturing, characterization and archiving of all cell lines. The Administrative Core provides centralized fiscal oversight and supports regular weekly Center meeting and the annual visits of the External Advisory Committee.

描述（由申请人提供）： 弗吉尼亚大学尤德尔帕金森中心检验的主要假设是，线粒体基因组突变对散发性帕金森病 (PD) 的发病机制有重大贡献，这种疾病困扰着 >95% 的帕金森病患者。在第一个资助期间，在证明血小板线粒体 DNA (mtDNA) 衍生的细胞杂种方面取得了实质性进展，即 PD 线粒体基因组的表达再现了 PD 的多种重要病理生物学特征。最引人注目的是，PD 杂种自发产生路易包涵体，其在生化和形态学上与 PD 脑中存在的包涵体无法区分。难以接受线粒体基因组对帕金森病的贡献，部分原因在于缺乏帕金森病大脑中尚未定义的特定线粒体DNA突变，担心帕金森病血小板线粒体DNA与脑神经元事件的相关性，无法提供线粒体DNA突变因果关系的直接证据缺乏对 mtDNA 突变导致 PD 中生物能缺陷和蛋白酶体功能障碍的了解。所有这些问题都在本续签申请中得到系统、全面的解决。 Bennett 博士的项目利用 UVA 创建的新技术，允许将来自任何来源（包括死后大脑）的人类线粒体基因组插入线粒体并在细胞中表达。将创建表达来自已明确表征的 PD 大脑的 mtDNA 的细胞系，并进行表型分析，从而可以直接测试因果关系。突变也可以通过诱变产生并在细胞中表达，直接证明线粒体DNA的致病性。 Capaldi 博士的项目涉及与俄勒冈大学的合作，对 PD 大脑线粒体中复合物 I 大分子组装和氧化损伤（与来自同一 PD 大脑表达 mtDNA 的细胞的线粒体进行比较）进行详细的蛋白质组学分析。项目 3 将揭示 PD mtDNA 突变引起的生物能复合物 I 损伤的机制。 Trimmer 博士的项目检查了 PD cybrids 单个细胞和 PD 大脑产生的细胞系中蛋白酶体损伤的出现，并定义了蛋白酶体损伤如何与致病蛋白聚集体的发育相关。该项目特别将 PD mtDNA 表达与神经元 PD 病理学联系起来。所有项目均由两个核心支持。细胞培养核心提供所有细胞系的集中培养、表征和存档。行政核心提供集中的财政监督，并支持每周定期的中心会议和外部咨询委员会的年度访问。