دورية أكاديمية
A high-throughput screening system for SARS-CoV-2 entry inhibition, syncytia formation and cell toxicity
| العنوان: | A high-throughput screening system for SARS-CoV-2 entry inhibition, syncytia formation and cell toxicity |
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| المؤلفون: | Shine Varghese Jancy, Santhik Subhasingh Lupitha, Aneesh Chandrasekharan, Shankara Narayanan Varadarajan, Shijulal Nelson-Sathi, Roshny Prasad, Sara Jones, Sreekumar Easwaran, Pramod Darvin, Aswathy Sivasailam, Thankayyan Retnabai Santhoshkumar |
| المصدر: | Biological Procedures Online, Vol 25, Iss 1, Pp 1-15 (2023) |
| بيانات النشر: | BMC, 2023. |
| سنة النشر: | 2023 |
| المجموعة: | LCC:Medicine (General) LCC:Biology (General) |
| مصطلحات موضوعية: | Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), VSV-eGFP-SARS-CoV-2, Human angiotensin-converting enzyme 2, Biosafety level 2 (BSL-2), High-throughput drug screening, Reporter assay system, Syncytia, Molecular simulation, Medicine (General), R5-920, Biology (General), QH301-705.5 |
| الوصف: | Abstract Background The entry of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) into the host cell is mediated through the binding of the SARS-CoV-2 Spike protein via the receptor binding domain (RBD) to human angiotensin-converting enzyme 2 (hACE2). Identifying compounds that inhibit Spike-ACE2 binding would be a promising and safe antiviral approach against COVID-19. Methods In this study, we used a BSL-2 compatible replication-competent vesicular stomatitis virus (VSV) expressing Spike protein of SARS-CoV-2 with eGFP reporter system (VSV-eGFP-SARS-CoV-2) in a recombinant permissive cell system for high-throughput screening of viral entry blockers. The SARS-CoV-2 permissive reporter system encompasses cells that stably express hACE2-tagged cerulean and H2B tagged with mCherry, as a marker of nuclear condensation, which also enables imaging of fused cells among infected EGFP positive cells and could provide real-time information on syncytia formation. Results A limited high-throughput screening identified six natural products that markedly inhibited VSV-eGFP-SARS-CoV-2 with minimum toxicity. Further studies of Spike-S1 binding using the permissive cells showed Scillaren A and 17-Aminodemethoxygeldanamycin could inhibit S1 binding to ACE2 among the six leads. A real-time imaging revealed delayed inhibition of syncytia by Scillaren A, Proscillaridin, Acetoxycycloheximide and complete inhibition by Didemnin B indicating that the assay is a reliable platform for any image-based drug screening. Conclusion A BSL-2 compatible assay system that is equivalent to the infectious SARS-CoV-2 is a promising tool for high-throughput screening of large compound libraries for viral entry inhibitors against SARS-CoV-2 along with toxicity and effects on syncytia. Studies using clinical isolates of SARS-CoV-2 are warranted to confirm the antiviral potency of the leads and the utility of the screening system. |
| نوع الوثيقة: | article |
| وصف الملف: | electronic resource |
| اللغة: | English |
| تدمد: | 1480-9222 |
| العلاقة: | https://doaj.org/toc/1480-9222Test |
| DOI: | 10.1186/s12575-023-00214-1 |
| الوصول الحر: | https://doaj.org/article/1349fc4b8aee440e8289ad842f26392fTest |
| رقم الانضمام: | edsdoj.1349fc4b8aee440e8289ad842f26392f |
| قاعدة البيانات: | Directory of Open Access Journals |
Full Text Finder | تدمد: | 14809222 |
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| DOI: | 10.1186/s12575-023-00214-1 |