Hyperoxia impairs intraflagellar transport and causes dysregulated metabolism with resultant decreased cilia length

Supplemental oxygen is a lifesaving measure in infants born premature to facilitate oxygenation. Unfortunately, it may lead to al-veolar simplification and loss of proximal airway epithelial cilia. Little is known about the mechanism by which hyperoxia causes ciliary dysfunction in the proximal respiratory tract. We hypothesized that hyperoxia causes intraflagellar transport (IFT) dysfunc-tion with resultant decreased cilia length. Differentiated basal human airway epithelial cells (HAEC) were exposed to hyperoxia or air for up to 48 h. Neonatal mice (<12 h old) were exposed to hyperoxia for 72 h and recovered in room air until postnatal day (PND) 60. Cilia length was measured from scanning electron microscopy images using a MATLAB-derived program. Proteomics and metabolomics were carried out in cells after hyperoxia. After hyperoxia, there was a significant time-dependent reduction in cilia length after hyperoxia in HAEC. Proteomic analysis showed decreased abundance of multiple proteins related to IFT including dynein motor proteins. In neonatal mice exposed to hyperoxia, there was a significant decrease in acetylated a tubulin at PND10 followed by recovery to normal levels at PND60. In HAEC, hyperoxia decreased the abundance of multiple proteins associated with complex I of the electron transport chain. In HAEC, hyperoxia increased levels of malate, fumarate, and citrate, and reduced the ATP/ADP ratio at 24 h with a subsequent increase at 36 h. Exposure to hyperoxia reduced cilia length, and this was associated with aberrant IFT protein expression and dysregulated metabolism. This suggests that hyperoxic expo-sure leads to aberrant IFT protein expression in the respiratory epithelium resulting in shortened cilia.

Automated summary

Supplemental oxygen is important for premature infants, but it may cause alveolar simplification and loss of airway epithelial cilia. The mechanism by which hyperoxia leads to ciliary dysfunction is not well understood. This study found that hyperoxia causes intraflagellar transport dysfunction, resulting in decreased cilia length. Proteomic analysis revealed aberrant expression of multiple proteins related to intraflagellar transport, and metabolomic analysis showed dysregulated metabolism in response to hyperoxia.