Donor-derived airway air-liquid interface model for high-throughput screening of antiviral combinations with concurrent analysis of antiviral efficacy and epithelial toxicity using ciliR.

BACKGROUND

Respiratory RNA viral infections significantly impact quality of life and productivity, in terms of both pandemics (e.g. severe acute respiratory syndrome coronavirus 2, influenza A viruses) and seasonal infections, such as respiratory syncytial virus (RSV). Globally, there are an estimated 33.8 million cases of RSV annually in children aged <5 years, leading to 2.8-4.3 million hospital admissions and up to 199 000 deaths. Human-relevant drug screening models are key to addressing the urgent need for effective antiviral treatments for RSV, particularly in vulnerable patient groups.

METHODS

Here, we present a donor-derived differentiated primary human nasal epithelial cell, high-throughput screening (HTS) Transwell-96 air-liquid interface (ALI) model designed to investigate the effects of combination antiviral therapies on RSV infection in primary human ciliated airway epithelium. In addition, we describe novel analytical tools using R (ciliR) to screen drug combinations by concurrently measuring efficacy and ciliary beat frequency, which we used as a sensitive marker of cell toxicity.

RESULTS

Our results demonstrate that the higher-throughput HTS Transwell-96 ALI cultures retain epithelial composition and ciliary function consistent within donor replicates. These cultures are permissible to infection with an RSV-green fluorescent protein (GFP) reporter virus, enabling quantitative comparison and combination treatment across multiple epithelial cultures from the same donor.

CONCLUSION

We anticipate that our disease-relevant system will serve as a foundation for larger-scale experiments aimed at optimising combination therapy for RSV and other respiratory viruses.