Bogetofte Helle, Jensen Pia, Ryding Matias, Schmidt Sissel I, Okarmus Justyna, Ritter Louise, Worm Christina S, Hohnholt Michaela C, Azevedo Carla, Roybon Laurent, Bak Lasse K, Waagepetersen Helle, Ryan Brent J, Wade-Martins Richard, Larsen Martin R, Meyer Morten
Department of Neurobiology Research, Institute of Molecular Medicine, University of Southern Denmark, Odense, Denmark.
Oxford Parkinson's Disease Centre, Medical Sciences Division, Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, United Kingdom.
Front Cell Neurosci. 2019 Jul 5;13:297. doi: 10.3389/fncel.2019.00297. eCollection 2019.
The protein parkin, encoded by the gene, is vital for mitochondrial homeostasis, and although it has been implicated in Parkinson's disease (PD), the disease mechanisms remain unclear. We have applied mass spectrometry-based proteomics to investigate the effects of parkin dysfunction on the mitochondrial proteome in human isogenic induced pluripotent stem cell-derived neurons with and without knockout (KO). The proteomic analysis quantified nearly 60% of all mitochondrial proteins, 119 of which were dysregulated in neurons with KO. The protein changes indicated disturbances in oxidative stress defense, mitochondrial respiration and morphology, cell cycle control, and cell viability. Structural and functional analyses revealed an increase in mitochondrial area and the presence of elongated mitochondria as well as impaired glycolysis and lactate-supported respiration, leading to an impaired cell survival in PARK2 KO neurons. This adds valuable insight into the effect of parkin dysfunction in human neurons and provides knowledge of disease-related pathways that can potentially be targeted for therapeutic intervention.
由该基因编码的蛋白质帕金对线粒体稳态至关重要，尽管它与帕金森病（PD）有关，但其发病机制仍不清楚。我们应用基于质谱的蛋白质组学技术，研究了帕金功能障碍对有或无基因敲除（KO）的人同基因诱导多能干细胞衍生神经元中线粒体蛋白质组的影响。蛋白质组学分析定量了几乎所有线粒体蛋白质的60%，其中119种在基因敲除的神经元中表达失调。蛋白质变化表明氧化应激防御、线粒体呼吸和形态、细胞周期控制及细胞活力受到干扰。结构和功能分析显示线粒体面积增加且存在拉长的线粒体，同时糖酵解和乳酸支持的呼吸受损，导致PARK2基因敲除神经元的细胞存活能力受损。这为帕金功能障碍对人类神经元的影响提供了有价值的见解，并提供了可能成为治疗干预靶点的疾病相关途径的知识。
