Therapeutic Development Branch, National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, 20850, USA.
Cancer Data Science laboratory, National Cancer Institute, National Institutes of Health, Bethesda, MD, 20892, USA.
J Transl Med. 2024 Sep 2;22(1):816. doi: 10.1186/s12967-024-05551-7.
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.
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丧失和含错误折叠α-突触核蛋白（α-syn）的路易体蛋白聚集体的积累，这些聚集体以磷酸化形式存在。缺乏有效的药物筛选模型阻碍了 PD 的药物开发研究。然而，最近体外类脑器官的发展为评估减缓这种慢性疾病进展的治疗剂提供了新的机会。
在这项研究中，我们使用 3D 类脑器官模型来研究重新利用抗病毒药物 Tilorone 来阻止α-突触核蛋白病传播的潜力。我们使用共聚焦显微镜评估 Tilorone 对荧光标记的α-syn 预形成纤维（sPFF）摄取和 sPFF 诱导的细胞凋亡的影响。我们还通过免疫印迹分析 midbrain-like 类器官提取物，研究了 Tilorone 对致病性 sPFF 诱导的内源性α-syn 磷酸化的影响。此外，我们还进行了定量 RT-PCR 和 sPFF 处理类器官的蛋白质组学分析，以评估 Tilorone 治疗对 3D 类器官模型中组织平衡的全局影响。
Tilorone 抑制 sPFF 在小鼠原代神经元和人 midbrain-like 类器官中的摄取。Tilorone 还降低了致病性α-syn 纤维诱导的内源性α-syn 磷酸化，并减轻了 midbrain-like 类器官中α-syn 纤维诱导的细胞凋亡。纤维处理类器官的蛋白质组学分析显示，α-syn 纤维会导致脂质代谢失衡，而 Tilorone 治疗可以逆转这种失衡。鉴于其在临床中的安全性，Tilorone 可能进一步开发为治疗 PD 患者突触核蛋白病传播的治疗干预措施。
