PATHOGENIC MECHANISMS OF NEURONAL CELL DEATH IN PD

PD 神经细胞死亡的致病机制

• 批准号: 6618255
• 负责人: JEREMY B TUTTLE
• 金额: $ 23.19万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-01 至 2003-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6618255
• 关键词: Parkinson's disease; biological signal transduction; biotechnology; calcium flux; calcium ion; cell death; cell line; clinical research; disease /disorder etiology; disease /disorder model; electron transport; free radical oxygen; human subject; hybrid cells; laboratory rat; mitochondria; mitochondrial DNA; molecular pathology; neurons; neuropathology; oxidative stress; pathologic process; tissue /cell culture

This is the second Project in a four Project group which is examining multiple aspects of mitochondrial functioning and mitochondrial defects associated with Parkinson's Disease (PD), a major neurodegenerative disease afflicting at least 1 million Americans. Project 2 is focused upon the pathogenetic mechanisms leading to the progressive neuronal cell death of the neurons affected in PD. The study will be conducted using cybrid model neurons and primary neurons in culture. Cybrid model neurons are constructed from a clonal human neural cell line lacking mitochondrial DNA (mtDNA) and patient-derived mitochondria. The project will examine a single central hypotheses: Mitochondrial involvement in oxidative stress, Ca2+ homeostasis and regulatory signaling is linked to increased vulnerability in Parkinson's disease- derived cybrids (PD cybrids). Mitochondria play an important role in the maintenance play an important role in the maintenance and efficacy of the calcium signaling network, especially in neurons. This is reflected in specific morphological and functional relationships between mitochondria and other cellular elements in the network, including the endoplasmic reticulum. The calcium signaling network plays a critical role coordinating exocytotic secretion, cellular metabolism, gene expression and inter-organelle communication. Mitochondria also function as a defense against cytotoxic of excitotoxic events and are implicated as a critical source of apoptotic signals including release of cytochrome c. Experiments will investigate the relationship between altered vulnerability to cell death in the cybrids derived from patients with Parkinson's disease and altered mitochondrial functioning in the calcium signaling network by addressing several aims: 1) What quantitative relationship exists between defects in ETC complex I activity, Ca2+ homeostasis and vulnerability to cell death in cybrids? Does neuroprotection involve restoration in normal Ca2+ homeostasis? 2) Does the impact of long-term inhibition of ETC complex I activity differ from that of acute inhibition? With respect to Ca2+ homeostasis? Cell death? Generation of oxyradicals? Mitochondrial morphology and inter- organelle interactions? 3) How are morphological and functional relationships of mitochondrial of mitochondria to other organelles affected in the PD cybrids? 4) How is mitochondrial movement altered in PD cybrids and in primary neurons after chronic rotenone treatment? 5) How do the mitochondrial defects in PD cybrids affect lysosomal activity, protein ubiquitination, degradation and the formation of inclusions that resemble Lewy bodies? The results gained will reveal novel aspects of the probable pathogenesis of this important disease in a carefully controlled model systems and may reveal therapeutic targets.

这是四个项目组中的第二个项目，该项目组正在研究与帕金森病（PD）相关的线粒体功能和线粒体缺陷的多个方面，帕金森病是一种主要的神经退行性疾病，至少有100万美国人受到影响。项目2的重点是导致PD患者神经元进行性神经元细胞死亡的发病机制。该研究将使用胞质杂种模型神经元和培养的原代神经元进行。Cybrid模型神经元由缺乏线粒体DNA（mtDNA）和患者来源的线粒体的克隆人神经细胞系构建。该项目将研究一个单一的中心假设：线粒体参与氧化应激，Ca2+稳态和调节信号与帕金森病衍生的胞质杂交体（PD胞质杂交体）的脆弱性增加有关。线粒体在钙信号网络的维持和功效中起着重要的作用，尤其是在神经元中。这反映在线粒体和网络中其他细胞元件（包括内质网）之间的特定形态和功能关系中。钙信号网络在细胞外分泌、细胞代谢、基因表达和细胞器间通讯中起着重要的协调作用。线粒体也作为防御细胞毒性的兴奋性毒性事件，并牵连作为一个关键来源的细胞凋亡信号，包括释放细胞色素c。实验将通过解决以下几个目标来研究来源于帕金森病患者的胞质杂交体中细胞死亡的脆弱性改变与钙信号网络中线粒体功能改变之间的关系：1）ETC复合物I活性、Ca2+稳态和胞质杂交体中细胞死亡的脆弱性之间存在什么定量关系？神经保护是否包括恢复正常的Ca2+稳态？2)ETC复合物I活性的长期抑制的影响与急性抑制的影响是否不同？关于Ca2+稳态？细胞死亡？产生氧自由基？线粒体形态和细胞器间的相互作用？3)在PD胞质杂种中，线粒体与其他细胞器的形态和功能关系是如何受到影响的？4)慢性鱼藤酮治疗后，PD胞质杂种和原代神经元的线粒体运动是如何改变的？5)PD胞质杂交体中的线粒体缺陷如何影响溶酶体活性、蛋白质泛素化、降解和类似路易体的内含物的形成？所获得的结果将揭示这种重要疾病在仔细控制的模型系统中的可能发病机制的新方面，并可能揭示治疗靶点。