We propose to build upon our complementary expertise in cutting edge bio-imaging and phospho-proteomic analysis to develop and use innovative drug screening concepts that have not been applied previously on parasitic systems. The consortium is based on 6 interactive scientific work packages that together propose a dual strategy for anti-leishmanial drug development. First, we will utilize visual high-content screening to discover compounds capable to kill intracellular Leishmania amastigotes without deteriorating the host cell. This phenotype-based strategy relies on fluorescent parasites and macrophages as read-outs and will allow simultaneous assessment of anti-leishmanial activity and host cell toxicity under physiological conditions. Second, we will apply a target-based strategy utilizing recombinant Leishmania protein kinases for inhibitor identification and structure-guided drug design. The identification of appropriate target kinases, with only limited homology to their mammalian counterparts will rely on (i) in silico analysis by applying novel bioinformatic tools developed by consortium members, and (ii) in vitro assay based on their phospho-transferase activity towards recombinant Leishmania phospho-proteins.
The major objectives of this proposal are