title: Deep-sequencing revealing mutation dynamics in the miltefosine transporter gene in Leishmania infantum selected for miltefosine resistance.
abstract: Miltefosine is the first oral drug used in chemotherapy against leishmaniasis. In vitro studies found that resistance to miltefosine in Leishmania is often associated with the acquisition of point mutations in the miltefosine transporter, leading to a decrease in drug uptake. In this study, the dynamics of mutations upon miltefosine selection was studied by deep-sequencing of the miltefosine transporter gene. Deep-sequencing data revealed that no mutation was detected in the miltefosine transporter at sub-inhibitory concentrations of miltefosine. We show that the prevalence of mutated alleles was increasing when the drug pressure heightened, that more mutations were observed in highly resistant mutants, and that most mutations remained when parasites were cultured for a few passages in the absence of miltefosine.
authors.affiliation: Centre de Recherche en Infectiologie, CRCHU de Québec, 2705 Boul. Laurier, Québec, Qc, G1V 4G2, Canada.
authors.forename: Marie-Claude N
authors.initials: MC
authors.lastname: Laffitte
authors.affiliation: Centre de Recherche en Infectiologie, CRCHU de Québec, 2705 Boul. Laurier, Québec, Qc, G1V 4G2, Canada.
authors.forename: Philippe
authors.initials: P
authors.lastname: Leprohon
authors.affiliation: Centre de Recherche en Infectiologie, CRCHU de Québec, 2705 Boul. Laurier, Québec, Qc, G1V 4G2, Canada.
authors.forename: Danielle
authors.initials: D
authors.lastname: Légaré
authors.affiliation: Centre de Recherche en Infectiologie, CRCHU de Québec, 2705 Boul. Laurier, Québec, Qc, G1V 4G2, Canada. marc.ouellette@crchul.ulaval.ca.
authors.forename: Marc
authors.initials: M
authors.lastname: Ouellette
mesh_terms: D000483:Alleles; D000818:Animals; D000981:Antiprotozoal Agents / Q000494:pharmacology*; D059014:High-Throughput Nucleotide Sequencing; D006801:Humans; D018314:Leishmania infantum / Q000187:drug effects / Q000235:genetics*; D007896:Leishmaniasis / Q000188:drug therapy / Q000469:parasitology*; D026901:Membrane Transport Proteins / Q000235:genetics*; D010767:Phosphorylcholine / Q000031:analogs & derivatives* / Q000494:pharmacology; D017354:Point Mutation; D017422:Sequence Analysis, DNA
chemical_list: D000981:Antiprotozoal Agents; D026901:Membrane Transport Proteins; D010767:Phosphorylcholine; C039128:miltefosine
keywords: Deep-sequencing; Leishmania; Miltefosine transporter; Resistance