title: Experimental resistance to drug combinations in Leishmania donovani: metabolic and phenotypic adaptations.
abstract: Together with vector control, chemotherapy is an essential tool for the control of visceral leishmaniasis (VL), but its efficacy is jeopardized by growing resistance and treatment failure against first-line drugs. To delay the emergence of resistance, the use of drug combinations of existing antileishmanial agents has been tested systematically in clinical trials for the treatment of visceral leishmaniasis (VL). In vitro, Leishmania donovani promastigotes are able to develop experimental resistance to several combinations of different antileishmanial drugs after 10 weeks of drug pressure. Using an untargeted liquid chromatography-mass spectrometry (LC-MS) metabolomics approach, we identified metabolic changes in lines that were experimentally resistant to drug combinations and their respective single-resistant lines. This highlighted both collective metabolic changes (found in all combination therapy-resistant [CTR] lines) and specific ones (found in certain CTR lines). We demonstrated that single-resistant and CTR parasite cell lines show distinct metabolic adaptations, which all converge on the same defensive mechanisms that were experimentally validated: protection against drug-induced and external oxidative stress and changes in membrane fluidity. The membrane fluidity changes were accompanied by changes in drug uptake only in the lines that were resistant against drug combinations with antimonials, and surprisingly, drug accumulation was higher in these lines. Together, these results highlight the importance and the central role of protection against oxidative stress in the different resistant lines. Ultimately, these phenotypic changes might interfere with the mode of action of all drugs that are currently used for the treatment of VL and should be taken into account in drug development.
authors: Unit of Molecular Parasitology, Department of Biomedical Sciences, Institute of Tropical Medicine, Antwerp, Belgium.
authors: Maya
authors: M
authors: Berg
authors: Instituto de Parasitología y Biomedicina López-Neyra, CSIC, Granada, Spain.
authors: Raquel
authors: R
authors: García-Hernández
authors: Unit of Molecular Parasitology, Department of Biomedical Sciences, Institute of Tropical Medicine, Antwerp, Belgium Biomedical Informatics Research Center Antwerp (Biomina), University of Antwerp/Antwerp University Hospital, Antwerp, Belgium Department of Mathematics and Computer Science, University of Antwerp, Antwerp, Belgium Department of Biomedical Sciences, University of Antwerp, Antwerp, Belgium.
authors: Bart
authors: B
authors: Cuypers
authors: Unit of Molecular Parasitology, Department of Biomedical Sciences, Institute of Tropical Medicine, Antwerp, Belgium.
authors: Manu
authors: M
authors: Vanaerschot
authors: Instituto de Parasitología y Biomedicina López-Neyra, CSIC, Granada, Spain.
authors: José I
authors: JI
authors: Manzano
authors: IBMC-Universidad Miguel Hernández, Elche (Alicante), Spain.
authors: José A
authors: JA
authors: Poveda
authors: IBMC-Universidad Miguel Hernández, Elche (Alicante), Spain.
authors: José A
authors: JA
authors: Ferragut
authors: Instituto de Parasitología y Biomedicina López-Neyra, CSIC, Granada, Spain.
authors: Santiago
authors: S
authors: Castanys
authors: Unit of Molecular Parasitology, Department of Biomedical Sciences, Institute of Tropical Medicine, Antwerp, Belgium Department of Biomedical Sciences, University of Antwerp, Antwerp, Belgium gamarro@ipb.csic.es jcdujardin@itg.be.
authors: Jean-Claude
authors: JC
authors: Dujardin
authors: Instituto de Parasitología y Biomedicina López-Neyra, CSIC, Granada, Spain gamarro@ipb.csic.es jcdujardin@itg.be.
authors: Francisco
authors: F
authors: Gamarro
mesh_terms: D000222:Adaptation, Physiological; D000818:Animals; D000977:Antiparasitic Agents / Q000494:pharmacology*; D002462:Cell Membrane / Q000187:drug effects; D002851:Chromatography, High Pressure Liquid; D016054:DNA, Protozoan / Q000235:genetics; D004338:Drug Combinations; D004351:Drug Resistance / Q000187:drug effects* / Q000235:genetics; D007893:Leishmania donovani / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D013058:Mass Spectrometry; D008560:Membrane Fluidity / Q000187:drug effects; D055432:Metabolomics; D018384:Oxidative Stress / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D000977:Antiparasitic Agents; D016054:DNA, Protozoan; D004338:Drug Combinations; D017382:Reactive Oxygen Species
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