Laboratory of Molecular and Chemical Biology of Neurodegeneration, Brain Mind Institute, Faculty of Life Sciences, École Polytechnique Fédérale de Lausanne (EPFL), Lausanne, CH, USA.
J Neurochem. 2021 May;157(4):891-898. doi: 10.1111/jnc.15275. Epub 2021 Mar 22.
Alpha-synuclein oligomerization is one of the early events on the pathway to Lewy body formation. Therefore, interfering with this process holds tremendous potential for developing therapies that block α-Syn pathology formation and toxicity. The development of robust and reliable cellular models of alpha-synuclein oligomerization is one important step toward achieving this goal. Unlike α-Syn fibrils, which can be detected and labeled using multiple tools and validated antibodies, α-Syn oligomers are very difficult to differentiate from soluble monomeric α-Syn in cells. This has led to increased reliance on fusing fluorescent proteins or fragments thereof to α-Syn to develop assays and cellular models to investigate α-Syn oligomerization. We recently presented results that highlight the limitation of one of these assays, the α-Syn Bimolecular Fluorescence (BIFC) assay (Frey et al. 2020b). Our findings underscored the critical importance of characterizing and validating cellular models before their use in mechanistic studies or drug discovery studies. In this commentary, I present my response to Dr Tiago Outeiro's recent commentary on this work, expand on our previous discussions on the BIFC assay, and propose an integrated approach for the development characterization, validation, and improvements of cellular models of α-Syn oligomerization and aggregation. Having access to multiple well-characterized and validated cellular models is essential not only for advancing our understanding of the biology of α-Syn and PD but also to identify novel therapeutic targets and drugs that could be successfully developed into treatments for PD and synucleinopathies. The more reliable the models, the faster we are likely to achieve these goals.
α-突触核蛋白寡聚化是形成路易体的途径中的早期事件之一。因此，干扰这一过程对于开发阻止α-Syn 病理学形成和毒性的疗法具有巨大的潜力。开发强大而可靠的α-突触核蛋白寡聚化细胞模型是实现这一目标的重要步骤之一。与可以使用多种工具和经过验证的抗体检测和标记的α-Syn 纤维不同，α-Syn 寡聚体在细胞中非常难以与可溶性单体α-Syn 区分开来。这导致人们越来越依赖于将荧光蛋白或其片段融合到α-Syn 上来开发测定和细胞模型以研究α-Syn 寡聚化。我们最近提出的结果强调了其中一种测定方法（α-突触核蛋白双分子荧光（BIFC）测定法）的局限性（Frey 等人，2020b）。我们的发现强调了在将细胞模型用于机制研究或药物发现研究之前对其进行表征和验证的重要性。在这篇评论中，我对 Tiago Outeiro 博士最近对这项工作的评论做出了回应，扩展了我们之前关于 BIFC 测定法的讨论，并提出了一种综合方法来开发、表征、验证和改进α-突触核蛋白寡聚化和聚集的细胞模型。获得多个经过良好表征和验证的细胞模型不仅对于推进我们对α-Syn 和 PD 生物学的理解至关重要，而且对于确定新的治疗靶点和药物也至关重要，这些药物有可能成功开发为 PD 和突触核蛋白病的治疗方法。模型越可靠，我们就越有可能更快地实现这些目标。
