E Jones1; L FitzPatrick1; M Challinor2; E Corrie2; R Kelly1; D Simpson3; L Frost1; E Offer1;
1 Medicines Discovery Catapult, UK; 2 Medicine Discovery Catapult, UK; 3 James & Lillian Martin Stem Cell Facility, Sir William Dunn School of Pathology, University of Oxford, UK
AbstractThe translation of therapies targeting CNS disorders from preclinical drug discovery to the clinic is complex, with a high attrition rate often due to a lack of efficacy. Identification of efficacious therapeutics will require the evaluation of potential new disease pathways and different mechanisms of action to uncover novel disease targets. Neuroinflammation is proposed to play a major role in across the spectrum of neurodegenerative diseases, and neuroinflammatory mediators are receiving increasing interest as potential drug targets. Microglia, the resident immune cells of the CNS, are key mediators of neuroinflammation in the CNS. In recent years, there has been a significant effort directed towards developing human in vitro CNS cell models, with the aim to improve the understanding of disease mechanisms and to increase clinical translation, with more relevant models. Here, we demonstrate human iPSC-derived microglia are functional and respond to inflammatory stimuli. To examine the activation of neuroinflammation signaling pathways in these cells, lentiviral-based reporters were used to evaluate the two phases of NLRP3 inflammasome activation; the priming (signal 1) and activation (signal 2) phases. First, we show NFĸB reporter activation in real-time in response to inflammation and the priming (signal 1) of inflammasome in iPSC-microglia. A second reporter assay examining the formation of ASC-specks was employed to analyse activation of NLRP3 inflammasome. Together, these innovative tools provide an opportunity for novel drug discovery and to further understanding of microglia in neuroinflammation and CNS disease.