Dual Targeting of Monomeric Tau and α-Synuclein Aggregation: A New Multitarget Therapeutic Strategy for Neurodegeneration
Dual Targeting of Monomeric Tau and α-Synuclein Aggregation: A New Multitarget Therapeutic Strategy for Neurodegeneration
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DOI：
10.1021/acschemneuro.0c00281
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发表时间：
2020-07-15
影响因子：
5
通讯作者：
Peccati, Francesca
中科院分区：
文献类型：
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作者：
Gabr, Moustafa T.;Peccati, Francesca
开发有效的多靶点治疗策略对于面对神经退行性疾病的多方面性质至关重要。帕金森病 (PD) 和阿尔茨海默病 (AD) 是两种最常见的神经退行性疾病，它们具有错误折叠蛋白质聚集体积累的共同特征，分别是路易体 (LB) 和神经原纤维缠结 (NFT)。 Tau 蛋白和 α-突触核蛋白 (α-syn) 是 LB 和 NFT 的前体，已在体外和体内模型中证明了协同聚集和神经毒性。在此，我们首次验证了单体 tau 和 α-syn 聚集的双重靶向作为开发神经系统疾病多靶点疗法的有效平台。基于细胞荧光共振能量转移 (FRET) 的高通量筛选 tau 结合 asa 化合物，然后对命中化合物进行额外筛选，以确定其阻止 α-syn 聚集的能力，确定了 MG-2119 潜在先导化合物。使用细胞 FRET 测定、等温滴定量热法 (ITC)、表面等离子共振 (SPR) 和微尺度热泳 (MSH) 验证了 MG-2119 与单体 tau 的高结合亲和力。此外，硫黄素 T (ThT) 测定和动态光散射 (DLS) 测量表明，MG-2119 抑制 a-syn 聚集。有趣的是，MG-2119 能够以剂量依赖性方式挽救 SH-SY5Y 神经母细胞瘤细胞中 tau 和 α-syn 联合诱导的细胞毒性。单靶点 tau 和 α-syn 聚集抑制剂观察到不太明显的细胞拯救效果，显示了本研究中描述的多靶点方法的优越性。 MG-2119 令人满意的药代动力学特征和低毒性有望在未来优化开发神经系统疾病的潜在疗法。
Development of efficient multitargeted therapeutic strategies is crucial in facing the multifaceted nature of neurodegenerative diseases. Parkinson's disease (PD) and Alzheimer's disease (AD), the two most common neurodegenerative disorders, share a common hallmark of accumulation of misfolded protein aggregates which are Lewy bodies (LBs) and neurofibrillary tangles (NFTs), respectively. Tau protein and alpha-synuclein (alpha-syn), the precursors of LBs and NFTs, have demonstrated synergistic aggregation and neurotoxicity in both in vitro and in vivo models. Herein, we validate for the first time dual targeting of monomeric tau and alpha-syn aggregation as an efficient platform for development of multitarget therapeutics for neurological disorders. Cellular fluorescence resonance energy transfer (FRET)-based high-throughput screening for tau-binding asa compounds, followed by additional screening of the hits for their ability to impede alpha-syn aggregation identified MG-2119 potential lead. The high binding affinity of MG-2119 to monomeric tau was verified using cellular FRET assay, isothermal titration calorimetry (ITC), surface plasmon resonance (SPR), and microscale thermophoresis (MSH). Moreover, MG-2119 inhibited a-syn aggregation as revealed by thioflavin T (ThT) assay and dynamic light scattering (DLS) measurements. Interestingly, MG-2119 was capable of rescuing combined tau and alpha-syn-induced cytotoxicity in SH-SY5Y neuroblastoma cells in a dose-dependent manner. Less pronounced cell-rescuing effects were observed for single-targeted tau and alpha-syn aggregation inhibitors showcasing the superiority of the multitargeted approach described in this study. The satisfactory pharmacokinetic profile and low toxicity of MG-2119 hold promise for future optimization to develop potential therapeutics for neurological disorders.