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Updated: May 28, 2025
Early Viral Entry Assays for the Identification and Evaluation of Antiviral Compounds
Published on: October 29, 2015
Establishment of a Panel of Human Cell Lines to Identify Cellular Receptors Used by Enteroviruses to Infect Cells
Anastasiia O Sosnovtseva1,2, Thi Hoa Le3, Dmitry S Karpov1,2
1Center for Precision Genome Editing and Genetic Technologies for Biomedicine, Engelhardt Institute of Molecular Biology, Russian Academy of Sciences, 119991 Moscow, Russia.
Abstract:
Non-pathogenic natural and recombinant strains of human Enteroviruses are the subject of ongoing study with some strains having been approved for use as anticancer agents. The efficacy of oncolytic virotherapy depends upon identifying the receptor utilized by a specific strain for cell entry, and the presence of this receptor on the surface of cancer cells. Accordingly, a rapid and straightforward approach to determining the enteroviral receptors is necessary for developing an effective patient-specific, virus-based cancer therapy. To this end, we created a panel of seven lines with double knockouts on the background of the HEK293T cell line, which lacks the IFNAR1 gene. In these lines, the main viral receptor genes, including PVR, CXADR, CD55, ITGA2, SCARB2, ICAM1, and FCGRT, were knocked out using the CRISPR/Cas9 system. The panel of lines was validated on twelve different Enteroviruses types, providing a basis for studying the molecular mechanisms of enterovirus entry into cells, and for developing new therapeutic strains.
Insights
Researchers developed a cell line panel to identify enterovirus receptors for oncolytic virotherapy. This tool aids in developing personalized cancer treatments using engineered enteroviruses.
Area of Science:
- Virology
- Molecular Biology
- Cancer Therapy
Background:
- Non-pathogenic enteroviruses are explored for anticancer applications (oncolytic virotherapy).
- Therapeutic efficacy relies on understanding viral entry receptors on cancer cells.
- A method to quickly identify enterovirus receptors is crucial for patient-specific therapies.
Purpose of the Study:
- To create a versatile cell line panel for identifying enterovirus receptors.
- To facilitate the development of novel oncolytic virus therapies.
Main Methods:
- Generated seven HEK293T cell lines with double knockouts of major enterovirus receptor genes (PVR, CXADR, CD55, ITGA2, SCARB2, ICAM1, FCGRT).
- Utilized CRISPR/Cas9 gene-editing technology for targeted gene knockouts.
- Validated the cell line panel using twelve different enterovirus types.
Main Results:
- Successfully created a panel of engineered cell lines lacking key viral entry receptors.
- Demonstrated the panel's utility in studying enterovirus-host interactions.
- Provided a foundation for investigating viral entry mechanisms.
Conclusions:
- The developed cell line panel is a valuable tool for enterovirus research.
- This platform supports the advancement of oncolytic virotherapy and personalized cancer treatment strategies.
