Genetic Models to Probe the Role of Complex-1 Dysfunction in Neurologic Disease

描述（由申请人提供）：也许在对抗或治愈帕金森病（PD）的疾病修饰疗法的发展中最大的障碍是缺乏预测性动物模型。虽然家族性帕金森病已经指导了当前一代的小鼠模型，但迄今为止，它们还未能概括出人类疾病的基本特征。这些模型中最明显的缺失是黑质自发退化，以及a-突触核蛋白在非突触核蛋白基础小鼠中的积累。鉴于在翻译领域对目标识别和验证的脊椎动物模型的不可否认的需求，必须考虑新的模型。我们将注意力集中在特发性帕金森病的各个方面，并合理设计了一种新的这种疾病的小鼠模型。线粒体复合物-1缺陷是特发性帕金森病的常规报道。此外，Complex-1的化学抑制剂在啮齿类动物和非人灵长类动物中重现了黑质的特征性选择性损伤、a-突触核蛋白的沉积以及随后的帕金森病。甚至有一小部分人摄入了复合物-1毒素，随后在临床上表现为左旋多巴反应性帕金森病。大量的遗传学研究也同样暗示了家族性帕金森病基因与线粒体生物学的关系，这进一步提高了我们对线粒体功能障碍在帕金森病中起主要作用的兴趣。我们建议通过基因靶向复合物-1功能来表征一种新的PD模型。复合物-1是一个由超过45个亚基组成的大型完整膜复合物，其组装由几个专有的伴侣辅助。一个关键的问题是，在这50个基因中，哪一个是PD模型最合适的靶点。我们关注的是一种复合物-1基因，当人类缺乏这种基因时，会导致黑质严重退化，而所有帕金森病患者都是同一大脑区域受到影响。这些数据强烈表明，该基因是产生特发性帕金森病动物模型的理想靶点。本探索性研究计划的目的是在体外研究复合物-1功能障碍的下游分子后果，并在一种新的PD遗传小鼠模型中表征复合物-1缺乏的基本神经病理学特性。这项工作将决定我们的模型是否适合进一步的基础和转化工作，以了解和治疗帕金森病的过程。

DESCRIPTION (provided by applicant): Perhaps the greatest obstacle in the development of disease-modifying therapeutics to combat or cure Parkinson's disease (PD) is the lack of predictive animal models. While familial forms of PD have guided the current generation of mouse models, they have thus far failed to recapitulate essential features of the human condition. Most notably lacking from these models is the spontaneous degeneration of the substantia nigra, and the accumulation of a-synuclein in non-synuclein based mice. Given the undeniable need for vertebrate models for target identification and validation in the translational arena, new models must be considered. We have focused our attention toward aspects of idiopathic PD in our rational design of a novel mouse model of this disease. Deficiencies in mitochondrial Complex-1 are routinely reported in idiopathic PD. In addition, chemical inhibitors of Complex-1 reproduce the characteristic selective lesion of the substantia nigra, deposition of the a-synuclein protein, and subsequent parkinsonism in rodents and non-human primates. There is even a small population of humans who ingested a Complex-1 toxin and subsequently presented clinically with an L-DOPA responsive parkinsonism. Numerous genetic studies have likewise implicated familial PD genes in mitochondrial biology, further heightening our interest in the role that mitochondrial dysfunction plays as a primary event in PD. We propose to characterize a novel model of PD by genetically targeting Complex-1 function. Complex-1 is a large integral membrane complex comprised of over 45 subunits whose assembly is aided by several proprietary chaperones. A critical question is which of these ~50 genes would be the most suitable target for a model of PD. We have focused on a Complex-1 gene that, when deficient in humans, results in severe degeneration of the substantia nigra, the same brain region affected in all PD cases. These data strongly suggest this gene to be an ideal target for the generation of an idiopathic PD animal model. The goal of this exploratory research proposal is to examine the downstream molecular consequences of Compex-1 dysfunction in vitro and characterize the basic neuropathological properties of Complex-1 deficiency in a novel genetic mouse model of PD. This work will determine the suitability of our model for further basic and translational efforts to understand and treat the PD disease process. PUBLIC HEALTH RELEVANCE: A major obstacle in the generation of new therapeutics for the treatment or prevention of Parkinson's disease is the lack of suitable animal models of spontaneous, premature nigral degeneration accompanied by other classic features of idiopathic Parkinson's disease. Here we will examine the suitability of a novel genetic model of Parkinson's disease in neuronal cell culture and in mice in an effort to satisfy perhaps the greatest unmet need in the field.