Wellcome Trust Clinical PhD Programme at the University of Dundee: 'Dissecting folate metabolism in Trypanosoma brucei.' (360G-Wellcome-104242_Z_14_Z)

The aim of the project is to further dissect folate metabolism in T.brucei, the causative agent of human African trypanosomiasis (HAT). An understanding of folate transport, the distribution of intra-cellular folates and the essentiality of enzymes in folate metabolism will not only provide a better understanding of folate metabolism but also inform our understanding of anti- folate drug mode of action and mechanisms of resistance. Specifically, we aim to: • Assess the role of thymidine kinase (TK) in modulating sensitivity to anti-folates through gene deletion • Determine how folate and its derivatives are taken up and further metabolised, and identify the genes involved using an RNAi-knockdown approach • Determine the effect of anti-folates on intra- cellular metabolite distribution • Identify mechanisms of resistance to anti- folates using a genome-scale tetracycline inducible RNAi library screen and compare these to experimentally derived resistant lines Throughout the project we will target key enzymes in the folate metabolic pathway (e.g. methionine synthase, folylpolyglutamyl synthase) to determine their essentiality and their potential as therapeutic drug targets, by creating conditional knockouts and over-expressers.