Abstract

Inflammation is an essential host response to infection and injury, but unregulated inflammation is highly damaging to the host and must be limited by negative feedback signalling. Inflammasomes are intracellular protein complexes that control the production of the pro-inflammatory cytokines IL-1beta and IL-18 and a lytic cell death programme known as pyroptosis. Inflammasome signalling is thus an extremely inflammatory process and inflammasome-dependent inflammation is associated with the pathogenesis of many common diseases including arthritis, Alzheimer’s Disease and NASH. While clinical trials for NLRP3 inflammasome inhibitors were initiated in 2019, there remains a deficit in our knowledge of inflammasome regulation, that urgently needs to be addressed. The endogenous mechanisms that limit inflammasome activity are not understood and this research proposal will address this gap in our knowledge. Our novel preliminary data demonstrate that fever range temperatures can specifically limit inflammasome activity in mouse and human macrophages. Furthermore, we have identified that NLRP3 itself is highly sensitive to temperature and thus may function as a thermosensor. We will investigate the temperature-dependent mechanisms of inflammasome regulation using a range of pharmacological and biochemical approaches. We will employ cellular thermal shift assays and cutting-edge advanced quantitative mass spectrometry techniques to study NLRP3 protein-protein interactions as a function of temperature. Mutations in NLRP3 cause familial cold autoinflammatory syndrome (FCAS) but how cold temperature triggers NLRP3 activation in FCAS is not understood. We will develop FCAS cell models including patient-derived iPSCs that will allow us to characterise FACS NLRP3 using a range of methods. This innovative proposal addresses fundamental questions in the biology of inflammation and our insights will help to advance inflammasome targeted therapies for human health.