qHTS to Identify Inhibitors of Zika Virus

qHTS 识别寨卡病毒抑制剂

• 批准号: 10269711
• 负责人: Anton Simeonov
• 金额: $ 11.24万
• 依托单位: NATIONAL CENTER FOR ADVANCING TRANSLATIONAL SCIENCES
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10269711
• 关键词: 3-Dimensional; Biological Assay; Collection; Congenital Abnormality; Disease Outbreaks; Flavivirus; Guillain-Barré Syndrome; Human; Infection; Intervention; Link; Microcephaly; Modeling; Natural Products; Neonatal; Neurologic; Pathogenicity; Recombinants; Renilla Luciferases; Reporter; Retina; System; Tissues; Toxic effect; Vaccines; Vero Cells; Viral; Viral Cytopathogenic Effect; Virus; ZIKA; ZIKV infection; Zika Virus; arthropod-borne; bioprinting; global health emergency; in vivo; inhibitor/antagonist; interest; miniaturize; nervous system disorder; novel; novel therapeutics; prevent; small molecule; small molecule inhibitor; transmission process

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.

虽然寨卡病毒（ZIKV）爆发引发的全球卫生紧急情况与严重的新生儿神经系统疾病已经消退，但ZIKV在流行地区的持续传播仍然存在。因此，对阐明和开发针对ZIKV（一种节肢动物传播的黄病毒）的干预措施保持了兴趣。为了鉴定小分子抗寨卡病毒化合物，我们筛选了6,000种化合物，其中大部分来自天然产物，以确定它们阻断野生型ZIKV感染的能力。我们使用了在Vero细胞中进行的病毒致细胞病变效应（CPE）抑制试验，该试验经过优化并小型化至1536孔格式。使用重组寨卡病毒对从初步测定中鉴定的适当活性化合物进行一组正交测定，一种使用重组海肾荧光素酶报告基因测定，另一种使用新型mCherry报告基因系统。进一步评价了六种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性模型，以更好地复制人类靶向寨卡组织的体内感染。