Primary Supervisor: Prof. L. O’Neill, Trinity College Dublin, Ireland
Title: Unravelling the effect of Pneumovirus and Influenza virus components on innate immune cell metabolism.
Collaborators: Prof. R. Fouchier, ErasmusMC, Rotterdam, the Netherlands, Dr. K. Pardali ,AstraZeneca, Gothenburg, Sweden
Early Stage Researcher: TBA (add once Early Stage Researcher list final)
Innate immune cell activation causes a metabolic reprograming leading to accumulation of intracellular metabolites and modulation of inflammation, but the impact of viral infections on such process is poorly understood. The aim of this project is to analyse whether interaction with Pneumovirus and Influenza virus (IAV) proteins and RNA sensing through PRRs can alter the metabolic profile of innate immune cells, and to elucidate the downstream effector mechanisms being triggered. Ligands for multiple PRRs mimicking +RNA viruses, such as poly I:C and dsRNA, will be tested in this context. The outcome will be compared to the one of lipopolysaccharide (LPS)-Toll-like receptor (TLR)-4 binding, a well-known inducer of aerobic glycolysis in macrophages. Expression of metabolic enzymes and nutrient carriers will be analysed by qPCR and western blotting, with a particular focus on pyruvate kinase M2 isoform (PKM2) and stabilisation of hypoxia-induced factor-1 (HIF-1), key drivers of TLR4-induced metabolic reprogramming in macrophages. Levels of intracellular metabolites such as succinate and itaconate will be measured by mass spectrometry, while qPCR and ELISAs will be used to quantify the production of inflammatory cytokines. To determine metabolic reprogramming, ESR6 will use the Seahorse technology. At ErasmusMC and in collaboration with ESR5, ESR6 will further investigate the specific impact of IAV and Pneumovirus infections on the phenotype and the metabolic profile of innate immune cells using the combined expertise of ErasmusMC(virology) and Trinity College Dublin (immunometabolism), while at AstraZeneca ESR6 will use the expertise of AstraZeneca to determine metabolic reorganisation in response to these virus infections in primary epithelial cells and the role of kinases such as MEK/ERK in this context. Overall, this project will highlight the possible modulation of immune and epithelial cell metabolism by viruses as part of viral mechanism of action. The project may identify novel metabolic-related pathways to be targeted in viral infections.