A cell-based, high-throughput screen for small molecule regulators of hepatitis C virus replication
A cell-based, high-throughput screen for small molecule regulators of hepatitis C virus replication
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DOI：
10.1053/j.gastro.2006.10.032
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发表时间：
2007-01-01
期刊：
影响因子：
29.4
通讯作者：
Chung, Raymond T.
中科院分区：
文献类型：
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作者：
Kim, Sun Suk;Peng, Lee F.;Chung, Raymond T.
背景与目的：在慢性丙型肝炎病毒感染的患者中，只有一半的患者对聚乙二醇化干扰素和利巴韦林有持续的病毒学应答，这会导致许多副作用。因此，确定更有效和更好的耐受性药物是一个高度优先的问题。我们应用化学生物学来筛选调节丙型肝炎病毒的小分子。方法：我们首先为384孔微孔板格式优化了Huh7/RepFeo复制子细胞，并使用这条线利用自动化技术筛选了大量具有良好特征的已知生物活性化合物文库。在第一次筛选中确定了几个能够刺激或抑制丙型肝炎病毒复制的分子后，我们在第二次筛选中使用全长的丙型肝炎病毒复制子细胞系验证了我们的命中化合物。结果：我们鉴定并验证了一些抗病毒药物和前病毒药物，包括HMG-CoA还原酶抑制剂(抗病毒药物)和皮质类固醇药物(前病毒药物)。与皮质类固醇相关的复制增加的发现表明，这些药物直接促进病毒复制，而不是依赖于它们对免疫反应的抑制作用。与HMG-CoA还原酶抑制剂相关的抗病毒活性的发现暗示了脂代谢在病毒生命周期中的重要作用。结论：我们开发了一种简单、可重复性和可靠的基于细胞的高通量筛选检测系统，使用丙型肝炎病毒复制子模型来识别调节丙型肝炎病毒复制的小分子。这种方法不仅可以用来鉴定可能的抗病毒药物，还可以用来鉴定病毒复制的细胞调节因子。
Background & Aims: Only half of patients with chronic hepatitis C virus (HCV) infection experience sustained virologic response to pegylated-interferon and ribavirin, which cause numerous side effects. Thus, the identification of more effective and better tolerated agents is a high priority. We applied chemical biology to screen small molecules that regulate HCV. Methods: We first optimized the Huh7/RepFeo replicon cell fine for the 384-well microplate format and used this line to screen a large library of well-characterized, known biologically active compounds using automated technology. After identifying several molecules capable of either stimulating or inhibiting HCV replication in this primary screen, we then validated our hit compounds using a full-length HCV replicon cell line in secondary screens. Results: We identified and validated a number of antiviral and proviral agents, including HMG-CoA reductase inhibitors (antiviral) and corticosteroids (proviral). The finding of increased replication associated with corticosteroids suggests that these agents directly promote viral replication independent of their suppressive effects on the immune response. The finding of antiviral activity associated with the HMG-CoA reductase inhibitors implies an important role for lipid metabolism in the viral life cycle. Conclusions: We have developed a simple, reproducible, and reliable cell-based high-throughput screening assay system using an HCV replicon model to identify small molecules that regulate HCV replication. This method can be used to identify not only putative antiviral agents, but also cellular regulators of viral replication.