Visualising the effects of pollution nanoparticles on respiratory epithelial cells at air-liquid interface

Pollution nanoparticles, termed particulate matter (PM), carry enormous population health burden, through direct and indirect effects that are thought to involve oxidation and inflammation. However, currently there is no single imaging or biochemical technique available to unequivocally assign the exact timing and the (bio)-chemical effects of PM components, thus preventing the implementation of solid strategies for the mitigation of their deleterious effects. This proposal will establish the exact site, sequence and timings of PM interaction with human airway epithelial cell and organelles. By establishing the relationship between these events this work will pinpoint the crucial subcellular processes that lead to oxidative stress and inflammation both at a single cell level and in whole cell populations. We will develop protocols to assess localisation via analytical cryo-electron microscope (cryo-EM) and direct oxidation pathways via fluorescence lifetime imaging microscopy (FLIM) in primary human bronchial epithelial cells (HBECs) grown in submerged culture and at air-liquid interface (ALI), which is the only model that accurately reflects airway pathophysiology, for the first time.