title: In vitro selection of miltefosine resistance in promastigotes of Leishmania donovani from Nepal: genomic and metabolomic characterization.
abstract: In this study, we followed the genomic, lipidomic and metabolomic changes associated with the selection of miltefosine (MIL) resistance in two clinically derived Leishmania donovani strains with different inherent resistance to antimonial drugs (antimony sensitive strain Sb-S; and antimony resistant Sb-R). MIL-R was easily induced in both strains using the promastigote-stage, but a significant increase in MIL-R in the intracellular amastigote compared to the corresponding wild-type did not occur until promastigotes had adapted to 12.2 μM MIL. A variety of common and strain-specific genetic changes were discovered in MIL-adapted parasites, including deletions at the LdMT transporter gene, single-base mutations and changes in somy. The most obvious lipid changes in MIL-R promastigotes occurred to phosphatidylcholines and lysophosphatidylcholines and results indicate that the Kennedy pathway is involved in MIL resistance. The inherent Sb resistance of the parasite had an impact on the changes that occurred in MIL-R parasites, with more genetic changes occurring in Sb-R compared with Sb-S parasites. Initial interpretation of the changes identified in this study does not support synergies with Sb-R in the mechanisms of MIL resistance, though this requires an enhanced understanding of the parasite's biochemical pathways and how they are genetically regulated to be verified fully.
authors.affiliation: Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral Street, Glasgow, G4 0RE, UK.
authors.forename: C D
authors.initials: CD
authors.lastname: Shaw
authors.affiliation: Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral Street, Glasgow, G4 0RE, UK.
authors.forename: J
authors.initials: J
authors.lastname: Lonchamp
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Cambridge, UK.|College of Science, NUI Galway, Galway, Ireland.
authors.forename: T
authors.initials: T
authors.lastname: Downing
authors.affiliation: Department of Biomedical Sciences, Instituut voor Tropische Geneeskunde Nationalestraat, Antwerpen, Belgium.
authors.forename: H
authors.initials: H
authors.lastname: Imamura
authors.affiliation: Department of Genetics, University of Cambridge, Cambridge, UK.
authors.forename: T M
authors.initials: TM
authors.lastname: Freeman
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Cambridge, UK.
authors.forename: J A
authors.initials: JA
authors.lastname: Cotton
authors.affiliation: Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Cambridge, UK.
authors.forename: M
authors.initials: M
authors.lastname: Sanders
authors.affiliation: Department of Biomedical Sciences, University of Antwerp, Universiteitsplein 1, Antwerpen, Belgium.|Glasgow Polyomics, University of Glasgow, Glasgow.
authors.forename: G
authors.initials: G
authors.lastname: Blackburn
authors.affiliation: Department of Biomedical Sciences, Instituut voor Tropische Geneeskunde Nationalestraat, Antwerpen, Belgium.|Department of Biomedical Sciences, University of Antwerp, Universiteitsplein 1, Antwerpen, Belgium.
authors.forename: J C
authors.initials: JC
authors.lastname: Dujardin
authors.affiliation: BP Koirala Institute of Health Sciences, Dharan, Nepal.
authors.forename: S
authors.initials: S
authors.lastname: Rijal
authors.affiliation: BP Koirala Institute of Health Sciences, Dharan, Nepal.
authors.forename: B
authors.initials: B
authors.lastname: Khanal
authors.affiliation: Department of Genetics, University of Cambridge, Cambridge, UK.
authors.forename: C J R
authors.initials: CJ
authors.lastname: Illingworth
authors.affiliation: Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral Street, Glasgow, G4 0RE, UK.
authors.forename: G H
authors.initials: GH
authors.lastname: Coombs
authors.affiliation: Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral Street, Glasgow, G4 0RE, UK.
authors.forename: K C
authors.initials: KC
authors.lastname: Carter
mesh_terms: D000818:Animals; D000965:Antimony / Q000494:pharmacology; D000981:Antiprotozoal Agents / Q000494:pharmacology*; D004351:Drug Resistance; D005260:Female; D007893:Leishmania donovani / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism*; D007898:Leishmaniasis, Visceral / Q000469:parasitology; D050356:Lipid Metabolism; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D009154:Mutation; D009390:Nepal; D021261:Parasitic Sensitivity Tests; D010767:Phosphorylcholine / Q000031:analogs & derivatives* / Q000494:pharmacology
chemical_list: D000981:Antiprotozoal Agents; D026901:Membrane Transport Proteins; D010767:Phosphorylcholine; C039128:miltefosine; D000965:Antimony