Establish new model systems to understand human neutrophil biology

Neutrophils are essential for infection defense but also drive severe tissue pathology, including COPD or ARDS.

To understand how neutrophils interact with tissues to mediate pathology, it is essential to understand how neutrophil activation (sterile or pathogen-induced) under physiological conditions shapes their proteomes and defines what molecules they secrete.

Aim 1 will use high resolution mass spectrometry to comprehensively quantify how human primary neutrophils in physiological medium modify signalling pathways and secretomes in response to pathogens (E.Coli and Candida albicans) or inflammatory stimuli (fMLP).

These data will provide a quantitative understanding of the full protein repertoire of neutrophils as they respond to pathogens.

The data will give new insights about how neutrophils mediate inflammatory responses and will identify targets for intervention to prevent neutrophil mediated immune pathology.

Aim 2 is to develop an experimental platform to genetically modify human neutrophils, since a model allowing genetic dissection of human neutrophil function would be invaluable.

Neutrophils can be generated in vitro from human induced pluripotent stem cells (iPSCs) but robustness and authenticity of this model is not well established.

I will use mass spectrometry to compare proteomes of iPSC-derived neutrophils to assess how comparable they are to primary human neutrophils.

Human iPSC can be genetically modified and I will assess the impact of genetic disruption of myeloperoxidase and NADPH oxidase components in iPSC on neutrophil differentiation and function to validate the system.

These studies will establish the feasibility of using iPSC-derived neutrophils as a platform for genetic manipulation of human neutrophil function.