Mast cells (MCs) abound in all barrier tissues and play an important role in pathogenesis of many chronic inflammatory diseases including urticaria, asthma, polyposis and food allergy. We studied immune cells obtained from nasal polyps and found increased percentages of MC expressing IL-17RB (receptor for IL-25), identifying a potentially novel “pathogenic“ MC subpopulation. Our aim is to use an unbiased approach to confirm features of IL-17RB+ MCs and to comprehensively evaluate MC heterogeneity at the single cell transcriptomic and epigenetic levels. Identified MC subpopulations will be further researched using already obtained transcriptional data from chronic urticaria patients, healthy controls and other available datasets of patients with chronic inflammatory diseases. To characterise functions of IL-17RB+ MCs a model of human MCs overexpressing IL-17RB has been established and will be used to study the role of IL-25/IL17RB in MC activation. Although there are many therapies targeting immune cells, there are no current treatments that specifically target MCs or MC specific pathways.
Year 1: the student will use experimental techniques for analysis of human MC lines and primary MCs isolated from polyp tissues, including tissue digestion, cell culture and flow cytometry. Subpopulations will be quantified, phenotyped and cell sorted for single cell analysis (Chromium Single Cell Multiome ATAC + Gene Expression).
Year 2: the student will evaluate airway MC heterogeneity at the single cell transcriptomic and chromatin levels in nasal polyps (single cell data analysis, confirmation of findings in primary MC cells using flow cytometry, cell signalling, cell culture).
Year 3: identified MC subpopulations will be further studied at functional level using recombinant models and human primary cells in context of chronic inflammatory diseases using a range of molecular immunology methods (RT-PCR, gene cloning, lentiviral gene delivery, calcium signalling, western blot).