Regulatory B cell metabolism and ageing: Integrating experiment and data analysis

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Regulatory B-cells (Bregs) are an important component of immune-tolerance, they arenumerically and phenotypically altered with increasing age and in cancer1-3. Bregdifferentiation and function, unlike other B cell populations, requires oxidativephosphorylation and high mitochrondria capacity and we have shown that interferencewith these metabolic pathways abrogates Bregs function4. Declining mitochondrialquality and metabolic activity are associated with normal aging, correlating with thedeclining of the immune function5. Here we will investigate how age-relatedmitochondrial/metabolic changes in Bregs affect their differentiation into plasma cellsand their suppressive function. Aim-1. Define age-related metabolic changes in B-cellsand Bregs (Mauri-Bradford UCL). PBMC/splenocytes from 20 to 70+ year-old donors willbe stained with antibodies defining B-cell subsets and metabolic pathways will beanalysed by multiparameter flow cytometry. B-cell glycolysis and respiration rates willalso be assessed by seahorse. Aim-2. Define age-related changes in the Bregmetabolic transcriptome. (Fraternali-Ng King's). We will sort Bregs from thespleens/blood of young (20-25y/o) and aged (70+) (n=3) adults and perform singlecell-RNA-sequencing. We propose a multi-disciplinary approach to interrogate agerelatedchanges of Breg metabolism through analysing the generated scRNA-seqdatasets using state-of-the-art computational algorithms followed by experimentalvalidation. Descriptions of key Breg cellular/metabolomic states identified bycomputational methods will be substantiated, integrating different biologicalannotations (protein-protein interactions, pathways, metabolomics) (Fraternali-Ng's).We will also perturb gene/metabolite drivers with genetic/pharmacological approachesto recapitulate or rescue age-associated Breg deficits (Mauri-Bradford's). We anticipateimproved understanding of B cells, Breg metabolism and its deregulation in ageing.Significance: Bregs are an essential component of the immune system andunderstanding how changes in their metabolic requirements in ageing affects theirregulatory function is crucial to a full understanding of immune homeostasis in oldage.