Kim Beom Jin, Noh Hye Rin, Jeon Hyongjun, Park Sang Myun
Department of Pharmacology, Ajou University School of Medicine, Suwon 16499, Korea.
Center for Convergence Research of Neurological Disorders, Ajou University School of Medicine, Suwon 16499, Korea.
Exp Neurobiol. 2023 Jun 30;32(3):147-156. doi: 10.5607/en23007.
Parkinson's disease (PD) is characterized by the presence of α-synuclein (α-syn) inclusions in the brain and the degeneration of dopamine-producing neurons. There is evidence to suggest that the progression of PD may be due to the prion-like spread of α-syn aggregates, so understanding and limiting α-syn propagation is a key area of research for developing PD treatments. Several cellular and animal model systems have been established to monitor α-syn aggregation and propagation. In this study, we developed an model using A53T α-syn-EGFP overexpressing SH-SY5Y cells and validated its usefulness for high-throughput screening of potential therapeutic targets. Treatment with preformed recombinant α-syn fibrils induced the formation of aggregation puncta of A53T α-syn-EGFP in these cells, which were analyzed using four indices: number of dots per cell, size of dots, intensity of dots, and percentage of cells containing aggregation puncta. Four indices are reliable indicators of the effectiveness of interventions against α-syn propagation in a one-day treatment model to minimize the screening time. This simple and efficient model system can be used for high-throughput screening to discover new targets for inhibiting α-syn propagation.
帕金森病（PD）的特征是大脑中存在α-突触核蛋白（α-syn）包涵体以及产生多巴胺的神经元退化。有证据表明，PD的进展可能是由于α-syn聚集体的朊病毒样传播，因此理解和限制α-syn的传播是开发PD治疗方法的关键研究领域。已经建立了几种细胞和动物模型系统来监测α-syn的聚集和传播。在本研究中，我们使用过表达A53T α-syn-EGFP的SH-SY5Y细胞开发了一种模型，并验证了其在高通量筛选潜在治疗靶点方面的实用性。用预先形成的重组α-syn纤维进行处理可诱导这些细胞中A53T α-syn-EGFP聚集点的形成，使用四个指标对其进行分析：每个细胞的点数量、点的大小、点的强度以及含有聚集点的细胞百分比。在一天的治疗模型中，这四个指标是针对α-syn传播的干预措施有效性的可靠指标，可最大限度地缩短筛选时间。这种简单有效的模型系统可用于高通量筛选，以发现抑制α-syn传播的新靶点。
