Abstract
Influenza and other respiratory viruses present a significant threat to public health, national security, and the world economy, and can lead to the emergence of global pandemics such as from COVID-19. A barrier to the development of effective therapeutics is the absence of a robust and predictive preclinical model, with most studies relying on a combination of in vitro screening with immortalized cell lines and low-throughput animal models. Here, we integrate human primary airway epithelial cells into a custom-engineered 96-device platform (PREDICT96-ALI) in which tissues are cultured in an array of microchannel-based culture chambers at an air–liquid interface, in a configuration compatible with high resolution in-situ imaging and real-time sensing. We apply this platform to influenza A virus and coronavirus infections, evaluating viral infection kinetics and antiviral agent dosing across multiple strains and donor populations of human primary cells. Human coronaviruses HCoV-NL63 and SARS-CoV-2 enter host cells via ACE2 and utilize the protease TMPRSS2 for spike protein priming, and we confirm their expression, demonstrate infection across a range of multiplicities of infection, and evaluate the efficacy of camostat mesylate, a known inhibitor of HCoV-NL63 infection. This new capability can be used to address a major gap in the rapid assessment of therapeutic efficacy of small molecules and antiviral agents against influenza and other respiratory viruses including coronaviruses.
Highlights
Influenza and other respiratory viruses present a significant threat to public health, national security, and the world economy, and can lead to the emergence of global pandemics such as from COVID19
We explored the formation of a pseudostratified epithelium at an ALI using freshly harvested human bronchial epithelial cells from living research bronchoscopies, labeled DH01, observing robust establishment of airway tissues over periods of 5 weeks or longer in culture in the PREDICT96-ALI plates
To determine if the PREDICT96-ALI airway model can be used to evaluate the efficacy of potential antiviral therapeutics, we investigated the effect of the antiviral agent oseltamivir—the most commonly used clinical anti-influenza therapy— for its ability to reduce viral load in influenza A viruses (IAV)-inoculated PREDICT96-ALI airway tissue
Summary
Influenza and other respiratory viruses present a significant threat to public health, national security, and the world economy, and can lead to the emergence of global pandemics such as from COVID19. While the integration of human primary cells or stem cell-derived populations in a physiologically-relevant microenvironment represents a tremendous advance over the use of immortalized cell lines in principle, in practice the application of these systems in pharmacological research and development has been gated by several factors These limitations include low-throughput (ranging from single microchamber devices to 6 – 24-device configurations) and complex operation, a lack of relevant metrics for system assessment, the use of research-grade materials and components for platform construction, the lack of critical aspects of tissue or organ structure required to replicate infection, and most critically, an absence of confidence in the in vitro-in vivo correlation (IVIVC) for these technologies
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