qHTS to Identify Inhibitors of Zika Virus

尽管寨卡病毒(ZIKV)暴发造成的与严重的新生儿神经疾病相关的全球卫生紧急情况已经消退，但ZIKV在流行地区的持续传播仍然存在。因此，阐明和开发针对ZIKV的干预措施一直是人们的兴趣，ZIKV是一种节肢动物传播的黄病毒。为了鉴定抗寨卡病毒的小分子化合物，我们筛选了6000种化合物，其中大部分来自天然产品，以确定它们是否具有阻止野生型寨卡病毒感染的能力。我们使用了在Vero细胞中进行的病毒细胞病变(CPE)抑制试验，该试验经过优化并微型化至1536孔格式。从初步试验中鉴定出的合适的活性化合物接受了一组使用重组寨卡病毒的正交试验，其中一个使用了重组Renilla荧光素酶报告试验，另一个使用了新的mCherry报告系统。进一步评价了6个活性分子对其他黄病毒的抑制作用。最后，我们在3D生物打印的视网膜组织中开发了一种新的病毒致病模型，以更好地复制人类靶向寨卡病毒组织的体内感染。

Although the global health emergency posed by the Zika virus (ZIKV) outbreak associated with severe neonatal neurological conditions has subsided, the continued transmission of ZIKV in endemic regions remains. As such, there is maintained interest in elucidating and developing interventions against ZIKV, a arthropod-borne flavivirus. To identify small-molecule anti-Zika compounds, we screened a collection of 6,000 compounds, most derived from natural products, for their ability to block wild-type ZIKV infection. We used a viral cytopathic effect (CPE) inhibition assay conducted in Vero cells that was optimized and miniaturized to 1536-well format. Suitably active compounds identified from the primary assay were subjected to a panel of orthogonal assays using recombinant Zika viruses, one using a recombinant Renilla luciferase reporter assay, the other a novel mCherry reporter system. Six active molecules were further evaluated for their inhibitory effects against other flaviviruses. Lastly, we developed a novel viral pathogenicity model in a 3D-bioprinted retinal tissue to better replicate in vivo infections of targeted Zika tissues in humans.