Harding, A. T.; Crossen, A. J.; Reedy, J. L.; Basham, K. J.; Hepworth, O. W.; Zhang, Y.; Shah, V. S.; Brown Harding, H.; Surve, M. V.; Simaku, P.; Kwaku, G. N.; Nolling Jensen, K.; Otto, Y.; Ward, R. A.; Thompson, G. R.; Klein, B. S.; Rajagopal, J.; Sen, P.; Haber, A. L.; Vyas, J. M.

Respiratory fungal infections pose a significant threat to human health. Animal models do not fully recapitulate human disease, necessitating advanced models to study human-fungal pathogen interactions. In this study, we utilized primary human airway epithelial cells (hAECs) to recapitulate the lung environment in vitro and investigate cellular responses to two diverse, clinically significant fungal pathogens, Aspergillus fumigatus and Coccidioides posadasii. To understand the mechanisms of early pathogenesis for both fungi, we performed single-cell RNA sequencing of infected hAECs. Analysis revealed that both fungi induced cellular stress and cytokine production. However, the cell subtypes affected and specific pathways differed between fungi, with A. fumigatus and C. posadasii triggering protein-folding-related stress in ciliated cells and hypoxia responses in secretory cells, respectively. This study represents one of the first reports of single-cell transcriptional analysis of hAECs infected with either A. fumigatus or C. posadasii, providing a vital dataset to dissect the mechanism of disease and potentially identify targetable pathways.