帕金森病 (PD) 是一种神经退行性疾病，其特征是多巴胺能神经元的丧失和含有磷酸化形式的错误折叠 α-突触核蛋白 (α-syn) 的路易体蛋白聚集体的积累。缺乏有效的药物筛选模型阻碍了帕金森病的药物开发研究。然而，最近体外类脑类器官的发展为评估治疗药物以减缓这种慢性疾病的进展提供了新的机会。在这项研究中，我们使用 3D 类脑类器官模型来研究重新利用替洛隆的潜力，一种抗病毒药物，用于阻止 α-突触核蛋白病的传播。我们使用共聚焦显微镜评估了替洛隆对荧光标记的 α-syn 预制原纤维 (sPFF) 的摄取和 sPFF 诱导的细胞凋亡的影响。我们还通过免疫印迹中脑样类器官提取物检查了替洛隆对致病性 sPFF 诱导的内源 α-syn 磷酸化的影响。此外，对 sPFF 处理的类器官进行定量 RT-PCR 和蛋白质组学分析，以评估 Tilorone 治疗对 3D 类器官模型中组织稳态的整体影响。Tilorone 抑制小鼠原代神经元和人类中脑样类器官对 sPFF 的摄取。 Tilorone 还可减少致病性 α-syn 原纤维诱导的内源 α-syn 磷酸化，并减轻 α-syn 原纤维诱导的中脑样类器官细胞凋亡。原纤维处理的类器官的蛋白质组学分析揭示了 α-syn 原纤维对脂质稳态的显着改变，而 Tilorone 处理可逆转这种改变。鉴于其在临床上的安全性，Tilorone 可能会被进一步开发为一种治疗干预措施，以减轻 PD 患者中突触核蛋白病的传播。© 2024。这是美国政府的作品，在美国不受版权保护；外国版权保护可能适用。

Parkinson's disease (PD) is a neurodegenerative condition characterized by the loss of dopaminergic neurons and the accumulation of Lewy-body protein aggregates containing misfolded α-synuclein (α-syn) in a phosphorylated form. The lack of effective models for drug screens has hindered drug development studies for PD. However, the recent development of in vitro brain-like organoids provides a new opportunity for evaluating therapeutic agents to slow the progression of this chronic disease.In this study, we used a 3D brain-like organoid model to investigate the potential of repurposing Tilorone, an anti-viral drug, for impeding the propagation of α-synucleinopathy. We assessed the effect of Tilorone on the uptake of fluorescently labeled α-syn preformed fibrils (sPFF) and sPFF-induced apoptosis using confocal microscopy. We also examined Tilorone's impact on the phosphorylation of endogenous α-syn induced by pathogenic sPFF by immunoblotting midbrain-like organoid extracts. Additionally, quantitative RT-PCR and proteomic profiling of sPFF-treated organoids were conducted to evaluate the global impact of Tilorone treatment on tissue homeostasis in the 3D organoid model.Tilorone inhibits the uptake of sPFF in both mouse primary neurons and human midbrain-like organoids. Tilorone also reduces the phosphorylation of endogenous α-syn induced by pathogenic α-syn fibrils and mitigates α-syn fibril-induced apoptosis in midbrain-like organoids. Proteomic profiling of fibril-treated organoids reveals substantial alterations in lipid homeostasis by α-syn fibrils, which are reversed by Tilorone treatment. Given its safety profile in clinics, Tilorone may be further developed as a therapeutic intervention to alleviate the propagation of synucleinopathy in PD patients.© 2024. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.