The role of endogenous PINK1 and Parkin mutations in human dopaminergic neurons

内源性 PINK1 和 Parkin 突变在人类多巴胺能神经元中的作用

• 批准号: 219522511
• 负责人: Professorin Dr. Christine Klein
• 金额: --
• 依托单位: Institut für Neurogenetik
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2012
• 资助国家: 德国
• 起止时间: 2011-12-31 至 2014-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/219522511?language=en
• 关键词: role; endogenous; PINK1; Parkin; mutations

Parkinson disease (PD) is the second most common neurodegenerative disorder, characterized by the loss of dopaminergic neurons in the substantia nigra. A clinical syndrome resembling ‘idiopathic’ PD has been reported for recessively inherited Parkin- and PINK1-linked PD. Our overall hypothesis in this application is that impairment of mitochondrial function in vulnerable neurons plays a key role in the pathogenesis of PD and that restoration of mitochondrial function will result in neuroprotection. As the function of the encoded proteins has been mainly investigated in non-neuronal systems or through knockdown approaches, there is an imperative to examine the role of endogenous mutations in appropriate human-derived and biologically relevant cell models. The objective of this study is to generate and characterize pure populations of iPS cell-derived dopaminergic neurons by using a novel differentiation strategy. Parkin and PINK1 mutant neurons will be examined for toxicity and mitochondrial function and compared to controls. In order to more directly test whether the observed phenotypes are related to pathogenic mutations, we intend to examine rescue strategies. We will express PGC-1α, an important regulator of mitochondrial biogenesis, and employ genetic rescue using zinc finger technology that allows for selective repair of endogenous mutations.

帕金森病(PD)是第二常见的神经退行性疾病，其特征是黑质多巴胺能神经元的丢失。据报道，隐性遗传性帕金森病和PINK1连锁帕金森病患者的临床症状类似于特发性帕金森病。我们在这一应用中的总体假设是，脆弱神经元中线粒体功能的损害在帕金森病的发病机制中起着关键作用，线粒体功能的恢复将导致神经保护。由于编码蛋白的功能主要是在非神经系统中或通过基因敲除方法进行研究，因此有必要研究内源性突变在适当的人类来源和生物相关细胞模型中的作用。本研究的目的是通过一种新的分化策略来产生和鉴定iPS细胞来源的多巴胺能神经元的纯群体。Parkin和PINK1突变神经元将被检测毒性和线粒体功能，并与对照组进行比较。为了更直接地测试观察到的表型是否与致病突变有关，我们打算检查救援策略。我们将表达线粒体生物发生的重要调节因子pGC-1α，并使用锌指技术进行遗传拯救，该技术允许选择性修复内源突变。