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A Human In Vitro Model to Study Adenoviral Receptors and Virus Cell Interactions.
Raphael L Tsoukas1,2, Wolfram Volkwein3, Jian Gao1
1Virology and Microbiology, Center for Biomedical Education and Research (ZBAF), Witten/Herdecke University, 58453 Witten, Germany.
Cells
|March 10, 2022
Summary
Understanding adenovirus infection mechanisms is key for gene therapy. This study used a human knockout model to identify specific adenovirus receptors, revealing CD46 and CAR dependencies for different adenovirus types.
Area of Science:
- Virology
- Gene Therapy
- Molecular Biology
Background:
- Adenoviral gene delivery requires understanding virus-host interactions.
- Key adenoviral attachment proteins include CD46, CAR, and sialic acid.
- Previous receptor studies often used non-human cells, limiting direct human application.
Purpose of the Study:
- To systematically analyze adenovirus receptor usage in human cells.
- To compare the side-by-side receptor dependency of various adenoviruses.
- To identify suitable adenovirus serotypes for gene therapy applications.
Main Methods:
- Utilized a reporter gene-tagged adenovirus library.
- Employed CRISPR/Cas9 to create CD46 and CAR knockout A549 cells (human lung epithelial carcinoma line).
- Infected knockout cells with 22 luciferase-expressing adenoviruses from species B, C, D, and E.
Main Results:
- Identified CD46-dependent adenoviruses (e.g., HAdV-B16, -B21, -B50, -B34, -B35).
- Identified CAR-dependent adenoviruses (e.g., HAdV-C5, -E4).
- Demonstrated dual CAR and CD46 roles for HAdV-B3, -B14, and species D viruses, suggesting additional receptors.
Conclusions:
- The human knockout cell model is effective for receptor usage analysis.
- This research aids in selecting appropriate adenoviruses for gene therapy.
- Provides insights into adenovirus infection biology and host cell interactions.
