Functions of the polyamine synthesis pathway in T cell biology (360G-Wellcome-201330_Z_16_Z)

Metabolism is essential for T cell fate and function. This proposal aims to investigate the role of an overlooked aspect of T cell metabolism, the polyamine synthesis pathway. The importance of polyamine metabolism in T cell proliferation, differentiation and function will be characterised using a conditional T cell murine knockout of the master regulator of polyamine synthesis (ODCflox/flox CD4-Cre). Where the polyamine pathway sits in the T cell metabolic network will be investigated in wildtype and ODC-/- T cells using various metabolomic techniques such as Seahorse, proteomics, and analysis of metabolite uptake/usage. How T cells utilise polyamines to drive cell division and differentiation will centre on the ability of polyamines to directly mediate epigenetic modifications and indirectly influence translation through the polyamine-dependent translation elongation factor eIF5a. Analysis of global acetylation/methylation patterns and gene expression in wildtype and ODC-/- T cells will provide insight as to how polyamines influence gene expression relevant to proliferation and differentiation. Similarly, how these molecules influence the translation of proteins also important for cell division and fate will be investigated through proteomic approaches and analysis of translation in ODC-/- and DOHH-/- (an enzyme required for eIF5a activation) T cells.