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Multiple Mutations in Heterogeneous Miltefosine-Resistant Leishmania major Population as Determined by Whole Genome Sequencing

BACKGROUND: Miltefosine (MF) is the first oral compound used in the chemotherapy against leishmaniasis. Since the mechanism of action of this drug and the targets of MF in Leishmania are unclear, we generated in a step-by-step manner Leishmania major promastigote mutants highly resistant to MF. Two...

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Autores principales: Coelho, Adriano C., Boisvert, Sébastien, Mukherjee, Angana, Leprohon, Philippe, Corbeil, Jacques, Ouellette, Marc
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2012
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3279362/
https://www.ncbi.nlm.nih.gov/pubmed/22348164
http://dx.doi.org/10.1371/journal.pntd.0001512
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author Coelho, Adriano C.
Boisvert, Sébastien
Mukherjee, Angana
Leprohon, Philippe
Corbeil, Jacques
Ouellette, Marc
author_facet Coelho, Adriano C.
Boisvert, Sébastien
Mukherjee, Angana
Leprohon, Philippe
Corbeil, Jacques
Ouellette, Marc
author_sort Coelho, Adriano C.
collection PubMed
description BACKGROUND: Miltefosine (MF) is the first oral compound used in the chemotherapy against leishmaniasis. Since the mechanism of action of this drug and the targets of MF in Leishmania are unclear, we generated in a step-by-step manner Leishmania major promastigote mutants highly resistant to MF. Two of the mutants were submitted to a short-read whole genome sequencing for identifying potential genes associated with MF resistance. METHODS/PRINCIPAL FINDINGS: Analysis of the genome assemblies revealed several independent point mutations in a P-type ATPase involved in phospholipid translocation. Mutations in two other proteins—pyridoxal kinase and α-adaptin like protein—were also observed in independent mutants. The role of these proteins in the MF resistance was evaluated by gene transfection and gene disruption and both the P-type ATPase and pyridoxal kinase were implicated in MF susceptibility. The study also highlighted that resistance can be highly heterogeneous at the population level with individual clones derived from this population differing both in terms of genotypes but also susceptibility phenotypes. CONCLUSIONS/SIGNIFICANCE: Whole genome sequencing was used to pinpoint known and new resistance markers associated with MF resistance in the protozoan parasite Leishmania. The study also demonstrated the polyclonal nature of a resistant population with individual cells with varying susceptibilities and genotypes.
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spelling pubmed-32793622012-02-17 Multiple Mutations in Heterogeneous Miltefosine-Resistant Leishmania major Population as Determined by Whole Genome Sequencing Coelho, Adriano C. Boisvert, Sébastien Mukherjee, Angana Leprohon, Philippe Corbeil, Jacques Ouellette, Marc PLoS Negl Trop Dis Research Article BACKGROUND: Miltefosine (MF) is the first oral compound used in the chemotherapy against leishmaniasis. Since the mechanism of action of this drug and the targets of MF in Leishmania are unclear, we generated in a step-by-step manner Leishmania major promastigote mutants highly resistant to MF. Two of the mutants were submitted to a short-read whole genome sequencing for identifying potential genes associated with MF resistance. METHODS/PRINCIPAL FINDINGS: Analysis of the genome assemblies revealed several independent point mutations in a P-type ATPase involved in phospholipid translocation. Mutations in two other proteins—pyridoxal kinase and α-adaptin like protein—were also observed in independent mutants. The role of these proteins in the MF resistance was evaluated by gene transfection and gene disruption and both the P-type ATPase and pyridoxal kinase were implicated in MF susceptibility. The study also highlighted that resistance can be highly heterogeneous at the population level with individual clones derived from this population differing both in terms of genotypes but also susceptibility phenotypes. CONCLUSIONS/SIGNIFICANCE: Whole genome sequencing was used to pinpoint known and new resistance markers associated with MF resistance in the protozoan parasite Leishmania. The study also demonstrated the polyclonal nature of a resistant population with individual cells with varying susceptibilities and genotypes. Public Library of Science 2012-02-14 /pmc/articles/PMC3279362/ /pubmed/22348164 http://dx.doi.org/10.1371/journal.pntd.0001512 Text en Coelho et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Coelho, Adriano C.
Boisvert, Sébastien
Mukherjee, Angana
Leprohon, Philippe
Corbeil, Jacques
Ouellette, Marc
Multiple Mutations in Heterogeneous Miltefosine-Resistant Leishmania major Population as Determined by Whole Genome Sequencing
title Multiple Mutations in Heterogeneous Miltefosine-Resistant Leishmania major Population as Determined by Whole Genome Sequencing
title_full Multiple Mutations in Heterogeneous Miltefosine-Resistant Leishmania major Population as Determined by Whole Genome Sequencing
title_fullStr Multiple Mutations in Heterogeneous Miltefosine-Resistant Leishmania major Population as Determined by Whole Genome Sequencing
title_full_unstemmed Multiple Mutations in Heterogeneous Miltefosine-Resistant Leishmania major Population as Determined by Whole Genome Sequencing
title_short Multiple Mutations in Heterogeneous Miltefosine-Resistant Leishmania major Population as Determined by Whole Genome Sequencing
title_sort multiple mutations in heterogeneous miltefosine-resistant leishmania major population as determined by whole genome sequencing
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3279362/
https://www.ncbi.nlm.nih.gov/pubmed/22348164
http://dx.doi.org/10.1371/journal.pntd.0001512
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