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

描述(申请人提供)：在开发疾病修正疗法来对抗或治愈帕金森氏病(PD)方面，也许最大的障碍是缺乏可预测的动物模型。虽然家族性帕金森病指导了当代的小鼠模型，但到目前为止，它们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的自发变性，以及非突触核蛋白基础的小鼠体内α-突触核蛋白的积累。鉴于在翻译领域对脊椎动物模型用于目标识别和验证的不可否认的需求，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，合理地设计了一种新的这种疾病的小鼠模型。线粒体复合体-1缺陷在特发性帕金森病中经常被报道。此外，复合体-1的化学抑制剂在啮齿动物和非人类灵长类动物中复制了典型的黑质选择性损伤、α-突触核蛋白沉积以及随后的帕金森氏症。甚至还有一小部分人摄入了Complex-1毒素，随后在临床上出现了L-多巴反应性帕金森综合症。大量的遗传学研究同样表明家族性帕金森病基因与线粒体生物学有关，这进一步增强了我们对线粒体功能障碍在帕金森病发病中的作用的兴趣。我们建议通过基因靶向Complex-1功能来描述一种新的帕金森病模型。复合体-1是一个由超过45个亚基组成的大型整体膜复合体，其组装由几个专有的伴侣辅助。一个关键的问题是，这50个基因中的哪一个是最适合帕金森病模型的靶点。我们专注于一种复杂-1基因，当人类缺乏该基因时，会导致黑质严重退化，这是所有帕金森病患者中受影响的同一大脑区域。这些数据强烈表明，该基因是建立特发性帕金森病动物模型的理想靶点。这项探索性研究计划的目标是在体外检测Compex-1功能障碍的下游分子后果，并在一种新的帕金森病遗传小鼠模型中表征Complex-1缺陷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于进一步了解和治疗帕金森病过程的基础和翻译工作。 公共卫生相关性：治疗或预防帕金森氏病的新疗法的产生的一个主要障碍是缺乏合适的动物模型，即伴随着特发性帕金森氏病的其他典型特征的自发性、早产性黑质退化。在这里，我们将检验一种新的帕金森氏病遗传模型在神经细胞培养和小鼠身上的适用性，以努力满足该领域可能最大的未得到满足的需求。

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.