In vitro selection of miltefosine resistance in promastigotes of Leishmania donovani from Nepal: genomic and metabolomic characterization

C. D. Shaw, J. Lonchamp, T. Downing, H. Imamura, T. M. Freeman, J. A. Cotton, M. Sanders, G. Blackburn, J. C. Dujardin, S. Rijal, B. Khanal, C. J. R. Illingworth, G. H. Coombs, K. C. Carter

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

n 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.
LanguageEnglish
Pages1134-1148
Number of pages15
JournalMolecular Microbiology
Volume99
Issue number6
Early online date9 Feb 2016
DOIs
Publication statusPublished - 1 Mar 2016

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miltefosine
Leishmania donovani
Nepal
Metabolomics
Parasites
Antimony
In Vitro Techniques
Lysophosphatidylcholines

Keywords

  • Leishmania donovani
  • miltefosine resistance
  • parasites

Cite this

Shaw, C. D. ; Lonchamp, J. ; Downing, T. ; Imamura, H. ; Freeman, T. M. ; Cotton, J. A. ; Sanders, M. ; Blackburn, G. ; Dujardin, J. C. ; Rijal, S. ; Khanal, B. ; Illingworth, C. J. R. ; Coombs, G. H. ; Carter, K. C. / In vitro selection of miltefosine resistance in promastigotes of Leishmania donovani from Nepal : genomic and metabolomic characterization. In: Molecular Microbiology. 2016 ; Vol. 99, No. 6. pp. 1134-1148.
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abstract = "n 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.",
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Shaw, CD, Lonchamp, J, Downing, T, Imamura, H, Freeman, TM, Cotton, JA, Sanders, M, Blackburn, G, Dujardin, JC, Rijal, S, Khanal, B, Illingworth, CJR, Coombs, GH & Carter, KC 2016, 'In vitro selection of miltefosine resistance in promastigotes of Leishmania donovani from Nepal: genomic and metabolomic characterization' Molecular Microbiology, vol. 99, no. 6, pp. 1134-1148. https://doi.org/10.1111/mmi.13291

In vitro selection of miltefosine resistance in promastigotes of Leishmania donovani from Nepal : genomic and metabolomic characterization. / Shaw, C. D.; Lonchamp, J.; Downing, T.; Imamura, H.; Freeman, T. M.; Cotton, J. A.; Sanders, M.; Blackburn, G.; Dujardin, J. C.; Rijal, S.; Khanal, B.; Illingworth, C. J. R.; Coombs, G. H.; Carter, K. C.

In: Molecular Microbiology, Vol. 99, No. 6, 01.03.2016, p. 1134-1148.

Research output: Contribution to journalArticle

TY - JOUR

T1 - In vitro selection of miltefosine resistance in promastigotes of Leishmania donovani from Nepal

T2 - Molecular Microbiology

AU - Shaw, C. D.

AU - Lonchamp, J.

AU - Downing, T.

AU - Imamura, H.

AU - Freeman, T. M.

AU - Cotton, J. A.

AU - Sanders, M.

AU - Blackburn, G.

AU - Dujardin, J. C.

AU - Rijal, S.

AU - Khanal, B.

AU - Illingworth, C. J. R.

AU - Coombs, G. H.

AU - Carter, K. C.

PY - 2016/3/1

Y1 - 2016/3/1

N2 - n 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.

AB - n 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.

KW - Leishmania donovani

KW - miltefosine resistance

KW - parasites

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