Primary Supervisor: Prof. J. Hiscott, Istituto Pasteur-Rome, Italy
Title: Effect of Influenza virus infection on Nrf2 activation of the oxidative stress response
Collaborators: Prof. A. Bowie, Trinity College Dublin, Ireland; Dr. N. Manel, STIMUNITY, Paris, France
Early Stage Researcher, TBA (add once Early Stage Researcher list final)
The ESR will investigate the influence of influenza virus infection and specific viral proteins on the role of oxidative stress responses in regulation of immune- and metabolic responses. These studies will focus on nuclear factor erythroid 2-related factor 2 (Nrf2), a global transcriptional regulator of the anti-oxidant gene network response, and the maintenance of redox homeostasis through the control of basal and induced expression of an array of antioxidant enzymes. It is our hypothesis that these redox regulatory checkpoints are activated by influenza infection to inhibit the innate antiviral response and to facilitate virus replication. Biochemical and molecular techniques, coupled with transcriptomic analysis and primary monocyte cell isolation will be used to measure ROS generation, mitochondrial depolarization and the generation of metabolites derived from the Krebs cycle, including succinate and itaconate, in terms of the reciprocal relationship between antioxidant Nrf2 and IFN regulatory networks. The effect of IAV infection, and individual viral proteins, on the overall metabolic profile of infected cells will be studied using Seahorse XF-24 technology. To investigate the mechanism of modulation in more detail, lung epithelial cells, macrophages and dendritic cells will be knocked-down for Nrf2 and other regulatory proteins, using CRISPr/Cas technology, and the consequence of ablation of Nrf2 on IAV replication kinetics, ROS production and metabolic profiles will be determined. These studies will provide: i) In depth understanding of the relationships between oxidative stress modulation, Nrf2-driven antioxidant responses and the regulation of the antiviral pathways directed against IAV infection; ii) Identification of metabolic products to modulate the Nrf2-IFN axis and impact on IAV pathogenesis. In collaboration with Dr. A. Bowie at TCD.