High-throughput drug screening using the Ebola virus transcription- and replication-competent virus-like particle system
| Autor: | Hichul Kim, Ji-Young Min, Honggun Lee, Thomas Hoenen, Jihye Lee, Nakyung Lee, Constantin Radu, Inhee Choi, Saehong Min, Jinyeong Heo, Yoonae Ko, Alexander König, Marc P. Windisch, David Shum |
|---|---|
| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Drug Cell Survival media_common.quotation_subject Drug Evaluation Preclinical Biology medicine.disease_cause Antiviral Agents 03 medical and health sciences Plasmid Therapeutic index Viral life cycle Transcription (biology) Virology medicine Humans Hit selection Viability assay Luciferases media_common Pharmacology Life Cycle Stages Neurotransmitter Agents Ebola virus Dose-Response Relationship Drug United States Food and Drug Administration Drug Repositioning Hemorrhagic Fever Ebola Ebolavirus United States High-Throughput Screening Assays HEK293 Cells 030104 developmental biology Linear Models Regression Analysis |
| Zdroj: | Antiviral Research. 158:226-237 |
| ISSN: | 0166-3542 |
| DOI: | 10.1016/j.antiviral.2018.08.013 |
| Popis: | The massive epidemic of Ebola virus disease (EVD) in West Africa, followed in recent months by two outbreaks in the Democratic Republic of the Congo, underline the importance of this severe disease. Because Ebola virus (EBOV) must be manipulated under biosafety level 4 (BSL4) containment, the discovery and development of virus-specific therapies have been hampered. Recently, a transient transfection-based transcription- and replication competent virus-like particle (trVLP) system was described, enabling modeling of the entire EBOV life cycle under BSL2 conditions. Using this system, we optimized the condition for bulk co-transfection of multiple plasmids, developed a luciferase reporter-based assay in 384-well microtiter plates, and performed a high-throughput screening (HTS) campaign of an 8,354-compound collection consisting of U.S. Food & Drug Administration (FDA) -approved drugs, bioactives, kinase inhibitors, and natural products in duplicates. The HTS achieved a good signal-to-background ratio with a low percent coefficient of variation resulting in Z’ = 0.7, and data points were reproducible with R2 = 0.89, indicative of a robust assay. After applying stringent hit selection criteria of ≥70% EBOV trVLP inhibition and ≥70% cell viability, 381 hits were selected targeting early, entry, and replication steps and 49 hits targeting late, maturation, and secretion steps in the viral life cycle. Of the total 430 hits, 220 were confirmed by dose-response analysis in the primary HTS assay. They were subsequently triaged by time-of-addition assays, then clustered and ranked according to their chemical structures, biological functions, therapeutic index, and maximum inhibition. Several novel drugs have been identified to very efficiently inhibit EBOV. Interestingly, most showed pharmacological activity in treatments for central nervous system-related diseases. We developed and screened an HTS assay using the novel EBOV trVLP system. Newly identified inhibitors are useful tools to study the poorly understood EBOV life cycle. In addition, they also provide opportunities to either repurpose FDA-approved drugs or develop novel viral interventions to combat EVD. |
| Databáze: | OpenAIRE |
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