Investigating the downstream effectors of the cAMP signalling pathway in Trypanosoma brucei as potential drug targets (360G-Wellcome-096984_Z_11_Z)

Current drug treatment options in trypanosomiasis are old, toxic and highly ineffective. Signalling transduction pathways have been shown to have important cellular functions and are therefore good pharmacological targets. The metabolism of cyclic adenosine monophosphate (cAMP) has recently been validated as a drug target in T. brucei. The phosphodiesterase inhibitor CpdA,used in the validation, was fatal to bloodstream forms. However, the downstream effectors of cAMP activity in trypanosomes are unknown. We intend to use reverse genetics, genomics, proteomics and metabolomics to elucidate potential pathways targeted by changes in cellular cAMP levels. Several genes have already been validated as reducing CpdA-sensitivity when knocked down through RNAi and we will study the expression, phosphorylation and localisation of the encoded proteins to understand their role in cAMP- regulated processes. We will try to identify further cellular components of the cAMP pathways by co-immunoprecipitation with these proteins, by comparative proteomics at various cAMP concentrations and by genomic analysis