Endothelial Inflammosome
Specific aims and working programme
The aim of this follow-up RTG project is to investigate the effect of metabolic stressors on the activation of the inflammasome in endothelial cells and to clarify the role of autophagy as an antiinflammatory mechanism contributing to resistance and persistence. Our hypothesis is that metabolic stressors stimulate an inflammatory response, which can be counteracted by simultaneously induced autophagy. We propose that balancing the inflammatory response by autophagy depends on the extent of metabolic stress and that either increasing stress or inhibiting autophagy may lead to cellular damage and cell death.
To address these questions we will investigate the dose- and time-dependent effects of metabolic stressors such as high glucose or palmitate on parameters of the NLRP3 inflammasome activation (expression of NLRPS, ASC and procaspase-1; caspase 1 activation, IL1b cleavage, ROS) and on autophagy induction (expression of autophagy proteins, LC3B conjugation, mitophagy markers). In addition, we will characterise how inhibition of autophagy (downregulation of autophagy proteins, PI3K inhibitors) affects the inflammatory response and explore the underlying mechanisms. Finally, we will apply activators of AMPK, which have been shown to stimulate autophagy in our previous studies, in vitro and in vivo and test their effects on inflammasome activation.