Table_1_PARK2 Mutation Causes Metabolic Disturbances and Impaired Survival of Human iPSC-Derived Neurons.XLSX

The protein parkin, encoded by the PARK2 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 PARK2 knockout (KO). The proteomic analysis quantified nearly 60% of all mitochondrial proteins, 119 of which were dysregulated in neurons with PARK2 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PARK2基因编码的帕金蛋白（parkin）对线粒体稳态至关重要，尽管该蛋白与帕金森病（PD）存在关联，但该病的具体发病机制仍未明确。本研究采用基于质谱的蛋白质组学技术，探究了帕金蛋白功能异常对人源同基因诱导多能干细胞（induced pluripotent stem cell, iPSC）分化所得神经元的线粒体蛋白质组的影响，实验设置了PARK2基因敲除（knockout, KO）组与野生型对照组。本次蛋白质组分析共定量了近60%的线粒体总蛋白，其中119种蛋白在PARK2基因敲除神经元中出现表达失调。上述蛋白表达变化提示，氧化应激防御、线粒体呼吸与形态调控、细胞周期调控以及细胞存活能力等多个通路均发生紊乱。结构与功能分析进一步显示，PARK2基因敲除神经元的线粒体面积增大、呈现细长形态，同时糖酵解及乳酸供能呼吸功能受损，最终导致细胞存活能力下降。本研究为阐明帕金蛋白功能异常对人神经元的影响提供了宝贵见解，并明确了可作为治疗干预靶点的疾病相关通路。