BBSRC EASTBIO PhD Programme: How is the activity of IL-33 controlled by the speed of its release, and how could this affect obesity?

This 4 year PhD project is part of a competition funded by EASTBIO BBSRC Doctoral Training Partnership http://www.eastscotbiodtp.ac.uk/how-apply-0. This opportunity is open to UK and EU nationals.

Applicants should apply by completing the EASTBIO application form (downloadable from the EASTBIO website) and e-mail to [Email Address Removed]. Candidates should also include their academic transcripts and ensure that they ask their referees to send completed references to [Email Address Removed]. Applicants may wish to explain their motivation for joining the EASTBIO training programme.

Supervisors: Dr Henry McSorley, Prof Simon Arthur (UoD) and Cecile Benezech (Centre for Cardiovascular Science, Edinburgh)

Obesity is associated with an inflammatory immune environment and contrasting suppression of type 2 immune responses. This project will investigate the role of the type 2-promoting cytokine IL-33 in obesity, using a range of parasite-derived protein modulators of the IL-33 pathway. Interleukin-33 is an alarmin cytokine –it is stored pre-formed, bound to genomic DNA in the nucleus of epithelial and stromal cells and released upon cell damage. It has an N-terminal DNA-binding domain, which prevents its release from the nucleus until this domain has been proteolytically cleaved. This binding to DNA appears to slow the release of IL-33 from the nucleus of necrotic epithelial cells, thus sustaining IL-33 signalling over longer enhancing its effects. This project will aim to study further how the speed of release of IL-33 controls the intensity of IL-33 responses. IL-33 is implicated in obesity, with IL-33-deficient mice becoming significantly more obese than wild-type mice. IL-33 release results in the activation of immune cells such as eosinophils and type 2 innate lymphoid cells which are known to be important in regulation of metabolism. We have previously shown the importance of expression of IL-33 by adipose tissue stromal cellsin control of local immune responses1. The effects of IL-33 can be modulated using HpARI, a parasite-derived protein that we identified, and which binds to IL-33 and DNA, blocking the effects of the cytokine2. We have developed a mutant version of HpARI, which has the surprising property of potently enhancing the effects of IL-33. It appears to act by affecting the speed and effective half-life of the cytokine’s release. We have also characterised elements of the signalling pathway which are required for productive IL-33 signalling, and which can be inhibited or exacerbated using small molecules3.

In this project, we will use our unique set of IL-33 modulators to determine how the speed of IL-33 release affects IL-33 responsiveness, and how this affects metabolic changes and obesity. We will carry out in vitro experiments in which we will measure the speed of release of IL-33 after treatment with HpARI mutants, using super-resolution live microscopy. We will assess how speed of release in culture affects downstream IL-33 signalling in IL-33-responsive cells such as ILC2s, mast cells, macrophagesand Th2 cells. Activation of signalling elements in responding cells will be assessed by flow cytometric staining, western blot, and proteomic analysis of global protein expression and modification.We will then apply these results to in vivo experiments administering mice with a high-fat diet to induce increases in white adipose fat accumulation, blocking the IL-33 pathway by using HpARI (or IL-33 receptor deficient mice), and amplifingIL-33 responses using HpARI mutants. We will assess weight gain, adipose fat accumulation, immune responsiveness, glucose resistance and insulin sensitivity. This project will therefore link a basic cellular process (release of nuclear proteins during cellular necrosis) and control of metabolism and obesity.