Human airway organoids as a versatile model to study BSL-4 virus replication and pathogenesis

Research with BSL-4 viruses such as Ebola, Marburg, and Nipah presents significant challenges due to their high virulence and the stringent containment measures required. A major limitation in studying viral pathogenesis and developing therapeutic strategies is the absence of suitable animal models that accurately replicate human disease. In this context, 3D cell culture systems offer significant advantages over traditional 2D monolayer cultures, mimicking native physiological conditions including cell polarization and composition. Human airway organoids, derived from pluripotent or adult stem cells, closely replicate the structure and function of the human respiratory system, providing a relevant and accessible environment for studying viral replication and pathogenesis. In contrast to conventional cell lines, airway organoids enable investigation of virus-host interactions within a human tissue context, providing insights that are more directly translatable to human disease. In our study, we generated airway organoids from both clinical donor tissues and commercially available nasal epithelial cells and showed in comparative analyses with whole lung tissue that these organoids are comparable in terms of cell composition. Despite donor-specific variations due to genetic factors, airway organoids derived from different sources and donors exhibit a remarkably similar cellular make-up. We further demonstrated that organoids derived from nasal swabs can effectively replicate BSL-4 viruses, establishing them as a standardized 3D model for broader research applications and advancing our understanding of these pathogens, especially in the absence of reliable animal models. Author Summary This study establishes human airway organoids as a robust model for investigating BSL-4 pathogens, such as Ebola, Marburg, and Nipah virus. Airway organoids represent reliable systems due to their ability to replicate the complexity of human respiratory epithelia and support viral infection. These organoids exhibit high susceptibility to these viruses, allowing for subsequent analysis of infection kinetics, immune evasion, and tissue-specific tropism within a controlled environment. This platform provides a powerful tool for antiviral testing and studying virus-host interactions, thus helping bridge critical gaps in high-containment virus research.