Amphotericin B resistance in Leishmania mexicana: Alterations to sterol metabolism and oxidative stress response.
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...
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Public Library of Science (PLoS)
2022-09-01
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| Series: | PLoS Neglected Tropical Diseases |
| Online Access: | https://journals.plos.org/plosntds/article/file?id=10.1371/journal.pntd.0010779&type=printable |
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| author | Edubiel A Alpizar-Sosa Nur Raihana Binti Ithnin Wenbin Wei Andrew W Pountain Stefan K Weidt Anne M Donachie Ryan Ritchie Emily A Dickie Richard J S Burchmore Paul W Denny Michael P Barrett |
| author_facet | Edubiel A Alpizar-Sosa Nur Raihana Binti Ithnin Wenbin Wei Andrew W Pountain Stefan K Weidt Anne M Donachie Ryan Ritchie Emily A Dickie Richard J S Burchmore Paul W Denny Michael P Barrett |
| author_sort | Edubiel A Alpizar-Sosa |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-d236c07b263b428594e99eab4248e1b2 |
| institution | Kabale University |
| issn | 1935-2727 1935-2735 |
| language | English |
| publishDate | 2022-09-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Neglected Tropical Diseases |
| spelling | doaj-art-d236c07b263b428594e99eab4248e1b22025-08-20T03:25:16ZengPublic Library of Science (PLoS)PLoS Neglected Tropical Diseases1935-27271935-27352022-09-01169e001077910.1371/journal.pntd.0010779Amphotericin B resistance in Leishmania mexicana: Alterations to sterol metabolism and oxidative stress response.Edubiel A Alpizar-SosaNur Raihana Binti IthninWenbin WeiAndrew W PountainStefan K WeidtAnne M DonachieRyan RitchieEmily A DickieRichard J S BurchmorePaul W DennyMichael P BarrettAmphotericin 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.https://journals.plos.org/plosntds/article/file?id=10.1371/journal.pntd.0010779&type=printable |
| spellingShingle | Edubiel A Alpizar-Sosa Nur Raihana Binti Ithnin Wenbin Wei Andrew W Pountain Stefan K Weidt Anne M Donachie Ryan Ritchie Emily A Dickie Richard J S Burchmore Paul W Denny Michael P Barrett Amphotericin B resistance in Leishmania mexicana: Alterations to sterol metabolism and oxidative stress response. PLoS Neglected Tropical Diseases |
| title | Amphotericin B resistance in Leishmania mexicana: Alterations to sterol metabolism and oxidative stress response. |
| title_full | Amphotericin B resistance in Leishmania mexicana: Alterations to sterol metabolism and oxidative stress response. |
| title_fullStr | Amphotericin B resistance in Leishmania mexicana: Alterations to sterol metabolism and oxidative stress response. |
| title_full_unstemmed | Amphotericin B resistance in Leishmania mexicana: Alterations to sterol metabolism and oxidative stress response. |
| title_short | Amphotericin B resistance in Leishmania mexicana: Alterations to sterol metabolism and oxidative stress response. |
| title_sort | amphotericin b resistance in leishmania mexicana alterations to sterol metabolism and oxidative stress response |
| url | https://journals.plos.org/plosntds/article/file?id=10.1371/journal.pntd.0010779&type=printable |
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