Division of Pharmacology and Pharmacotherapy, Faculty of Pharmacy, University of Helsinki, Finland.
Division of Pharmacology and Pharmacotherapy, Faculty of Pharmacy, University of Helsinki, Finland; School of Pharmacy, Faculty of Health Sciences, University of Eastern Finland, Finland.
Neuropharmacology. 2022 Nov 1;218:109213. doi: 10.1016/j.neuropharm.2022.109213. Epub 2022 Aug 12.
Parkinson's disease (PD) is characterized by degeneration of nigrostriatal dopaminergic neurons and accumulation of α-synuclein (αSyn) as Lewy bodies. Currently, there is no disease-modifying therapy available for PD. We have shown that a small molecular inhibitor for prolyl oligopeptidase (PREP), KYP-2047, relieves αSyn-induced toxicity in various PD models by inducing autophagy and preventing αSyn aggregation. In this study, we wanted to study the effects of PREP inhibition on different αSyn species by using cell culture and in vivo models. We used Neuro2A cells with transient αSyn overexpression and oxidative stress or proteasomal inhibition-induced αSyn aggregation to assess the effect of KYP-2047 on soluble αSyn oligomers and on cell viability. Here, the levels of soluble αSyn were measured by using ELISA, and the impact of KYP-2047 was compared to anle138b, nilotinib and deferiprone. To evaluate the effect of KYP-2047 on αSyn fibrillization in vivo, we used unilateral nigral AAV1/2-A53T-αSyn mouse model, where the KYP-2047 treatment was initiated two- or four-weeks post injection. KYP-2047 and anle138b protected cells from αSyn toxicity but interestingly, KYP-2047 did not reduce soluble αSyn oligomers. In AAV-A53T-αSyn mouse model, KYP-2047 reduced significantly proteinase K-resistant αSyn oligomers and oxidative damage related to αSyn aggregation. However, the KYP-2047 treatment that was initiated at the time of symptom onset, failed to protect the nigrostriatal dopaminergic neurons. Our results emphasize the importance of whole αSyn aggregation process in the pathology of PD and raise an important question about the forms of αSyn that are reasonable targets for PD drug therapy.
帕金森病（PD）的特征是黑质纹状体多巴胺能神经元退化和α-突触核蛋白（αSyn）聚集成路易体。目前，尚无针对 PD 的疾病修饰疗法。我们已经表明，脯氨酰寡肽酶（PREP）的小分子抑制剂 KYP-2047 通过诱导自噬和防止αSyn 聚集来缓解各种 PD 模型中的αSyn 诱导的毒性。在这项研究中，我们希望通过细胞培养和体内模型研究 PREP 抑制对不同αSyn 物种的影响。我们使用瞬时过表达αSyn 和氧化应激或蛋白酶体抑制诱导αSyn 聚集的 Neuro2A 细胞来评估 KYP-2047 对可溶性αSyn 寡聚物和细胞活力的影响。在这里，通过 ELISA 测量可溶性αSyn 的水平，并将 KYP-2047 的作用与 anle138b、nilotinib 和 deferiprone 进行比较。为了评估 KYP-2047 对体内αSyn 纤维化的影响，我们使用了单侧黑质 AAV1/2-A53T-αSyn 小鼠模型，其中在注射后两到四周开始 KYP-2047 治疗。KYP-2047 和 anle138b 可保护细胞免受αSyn 毒性，但有趣的是，KYP-2047 并未减少可溶性αSyn 寡聚物。在 AAV-A53T-αSyn 小鼠模型中，KYP-2047 显著减少了蛋白酶 K 抗性的αSyn 寡聚物和与αSyn 聚集相关的氧化损伤。然而，在症状出现时开始的 KYP-2047 治疗未能保护黑质纹状体多巴胺能神经元。我们的结果强调了整个αSyn 聚集过程在 PD 病理学中的重要性，并提出了一个关于 PD 药物治疗合理靶标形式的重要问题。
