K Hennig1, J Abi-Ghanem1, A Bunescu1, X Meniche1, E Biliaut1, A D Ouattara1, M D Lewis2, J M Kelly2, S Braillard3, G Courtemanche1, E Chatelain3, F Béquet4. 1. BIOASTER, 40 avenue Tony Garnier, 69007, Lyon, France. 2. Department of Pathogen Molecular Biology, London School of Hygiene and Tropical Medicine, Keppel Street, London, WC1E 7HT, UK. 3. Drugs for Neglected Diseases Initiative (DNDi), 15 Chemin Louis-Dunant, 1202, Geneva, Switzerland. 4. BIOASTER, 40 avenue Tony Garnier, 69007, Lyon, France. frederic.bequet@bioaster.org.
Abstract
INTRODUCTION: Chagas disease, the most important parasitic infection in Latin America, is caused by the intracellular protozoan Trypanosoma cruzi. To treat this disease, only two nitroheterocyclic compounds with toxic side effects exist and frequent treatment failures are reported. Hence there is an urgent need to develop new drugs. Recently, metabolomics has become an efficient and cost-effective strategy for dissecting drug mode of action, which has been applied to bacteria as well as parasites, such as different Trypanosome species and forms. OBJECTIVES: We assessed if the metabolomics approach can be applied to study drug action of the intracellular amastigote form of T. cruzi in a parasite-host cell system. METHODS: We applied a metabolic fingerprinting approach (DI-MS and NMR) to evaluate metabolic changes induced by six different (candidate) drugs in a parasite-host cell system. In a second part of our study, we analyzed the impact of two drugs on polar metabolites, lipid and proteins to evaluate if affected pathways can be identified. RESULTS: Metabolic signatures, obtained by the fingerprinting approach, resulted in three different clusters. Two can be explained by already known of mode actions, whereas the three experimental drugs formed a separate cluster. Significant changes induced by drug action were observed in all the three metabolic fractions (polar metabolites, lipids and proteins). We identified a general impact on the TCA cycle, but no specific pathways could be attributed to drug action, which might be caused by a high percentage of common metabolome between a eukaryotic host cell and a eukaryotic parasite. Additionally, ion suppression effects due to differences in abundance between host cells and parasites may have occurred. CONCLUSION: We validated the metabolic fingerprinting approach to a complex host-cell parasite system. This technique can potentially be applied in the early stage of drug discovery and could help to prioritize early leads or reconfirmed hits for further development.
INTRODUCTION: Chagas disease, the most important parasitic infection in Latin America, is caused by the intracellular protozoan Trypanosoma cruzi. To treat this disease, only two nitroheterocyclic compounds with toxic side effects exist and frequent treatment failures are reported. Hence there is an urgent need to develop new drugs. Recently, metabolomics has become an efficient and cost-effective strategy for dissecting drug mode of action, which has been applied to bacteria as well as parasites, such as different Trypanosome species and forms. OBJECTIVES: We assessed if the metabolomics approach can be applied to study drug action of the intracellular amastigote form of T. cruzi in a parasite-host cell system. METHODS: We applied a metabolic fingerprinting approach (DI-MS and NMR) to evaluate metabolic changes induced by six different (candidate) drugs in a parasite-host cell system. In a second part of our study, we analyzed the impact of two drugs on polar metabolites, lipid and proteins to evaluate if affected pathways can be identified. RESULTS: Metabolic signatures, obtained by the fingerprinting approach, resulted in three different clusters. Two can be explained by already known of mode actions, whereas the three experimental drugs formed a separate cluster. Significant changes induced by drug action were observed in all the three metabolic fractions (polar metabolites, lipids and proteins). We identified a general impact on the TCA cycle, but no specific pathways could be attributed to drug action, which might be caused by a high percentage of common metabolome between a eukaryotic host cell and a eukaryotic parasite. Additionally, ion suppression effects due to differences in abundance between host cells and parasites may have occurred. CONCLUSION: We validated the metabolic fingerprinting approach to a complex host-cell parasite system. This technique can potentially be applied in the early stage of drug discovery and could help to prioritize early leads or reconfirmed hits for further development.
Authors: Darren J Creek; Hwa H Chua; Simon A Cobbold; Brunda Nijagal; James I MacRae; Benjamin K Dickerman; Paul R Gilson; Stuart A Ralph; Malcolm J McConville Journal: Antimicrob Agents Chemother Date: 2016-10-21 Impact factor: 5.191
Authors: Kei Katsuno; Jeremy N Burrows; Ken Duncan; Rob Hooft van Huijsduijnen; Takushi Kaneko; Kiyoshi Kita; Charles E Mowbray; Dennis Schmatz; Peter Warner; B T Slingsby Journal: Nat Rev Drug Discov Date: 2015-10-05 Impact factor: 84.694
Authors: Shane R Wilkinson; Martin C Taylor; David Horn; John M Kelly; Ian Cheeseman Journal: Proc Natl Acad Sci U S A Date: 2008-03-26 Impact factor: 11.205
Authors: Mônica C Campos; Jody Phelan; Amanda F Francisco; Martin C Taylor; Michael D Lewis; Arnab Pain; Taane G Clark; John M Kelly Journal: Sci Rep Date: 2017-10-31 Impact factor: 4.379