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Amphotericin B resistance in Leishmania mexicana: Alterations to sterol metabolism and oxidative stress response

Alpizer-Sosa, E.A.; Ithnin, N.R.B.; Wei, W.; Pountain, A.W.; Weidt, A.K.; Donachie, A.M.; Ritchie, R.; Dickie, E.A; Burchmore, R.J.S.; Denny, P.W.; Barrett, M.P.

Amphotericin B resistance in Leishmania mexicana: Alterations to sterol metabolism and oxidative stress response Thumbnail


Authors

N.R.B. Ithnin

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Dr Wenbin Wei wenbin.wei2@durham.ac.uk
Chief Experimental Officer (Bioinformatics)

A.W. Pountain

A.K. Weidt

A.M. Donachie

R. Ritchie

E.A Dickie

R.J.S. Burchmore

M.P. Barrett



Abstract

Amphotericin B is increasingly used in treatment of leishmaniasis. Here, fourteen independent lines of Leishmania mexicana and one L. infantum line were selected for resistance to either amphotericin B or the related polyene antimicrobial, nystatin. Sterol profiling revealed that, in each resistant line, the predominant wild-type sterol, ergosta-5,7,24-trienol, was replaced by other sterol intermediates. Broadly, two different profiles emerged among the resistant lines. Whole genome sequencing then showed that these distinct profiles were due either to mutations in the sterol methyl transferase (C24SMT) gene locus or the sterol C5 desaturase (C5DS) gene. In three lines an additional deletion of the miltefosine transporter gene was found. Differences in sensitivity to amphotericin B were apparent, depending on whether cells were grown in HOMEM, supplemented with foetal bovine serum, or a serum free defined medium (DM). Metabolomic analysis after exposure to AmB showed that a large increase in glucose flux via the pentose phosphate pathway preceded cell death in cells sustained in HOMEM but not DM, indicating the oxidative stress was more significantly induced under HOMEM conditions. Several of the lines were tested for their ability to infect macrophages and replicate as amastigote forms, alongside their ability to establish infections in mice. While several AmB resistant lines showed reduced virulence, at least two lines displayed heightened virulence in mice whilst retaining their resistance phenotype, emphasising the risks of resistance emerging to this critical drug.

Citation

Alpizer-Sosa, E., Ithnin, N., Wei, W., Pountain, A., Weidt, A., Donachie, A., …Barrett, M. (2022). Amphotericin B resistance in Leishmania mexicana: Alterations to sterol metabolism and oxidative stress response. PLoS Neglected Tropical Diseases, 16(9), Article e0010779. https://doi.org/10.1371/journal.pntd.0010779

Journal Article Type Article
Acceptance Date Aug 31, 2022
Online Publication Date Sep 28, 2022
Publication Date 2022
Deposit Date Sep 7, 2022
Publicly Available Date Oct 21, 2022
Journal PLOS Neglected Tropical Diseases
Print ISSN 1935-2727
Electronic ISSN 1935-2735
Publisher Public Library of Science
Peer Reviewed Peer Reviewed
Volume 16
Issue 9
Article Number e0010779
DOI https://doi.org/10.1371/journal.pntd.0010779
Public URL https://durham-repository.worktribe.com/output/1194925

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Published Journal Article (2.7 Mb)
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Publisher Licence URL
http://creativecommons.org/licenses/by/4.0/

Copyright Statement
This is an open access article, free of all
copyright, and may be freely reproduced,
distributed, transmitted, modified, built upon, or
otherwise used by anyone for any lawful purpose.
The work is made available under the Creative
Commons CC0 public domain dedication.






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