Metabolomics and lipidomics reveal perturbation of sphingolipid metabolism by a novel anti-trypanosomal 3-(oxazolo[4,5-b]pyridine-2-yl)anilide

Daniel Stoessel, Cameron J. Nowell, Amy J. Jones, Lori Ferrins, Katherine M. Ellis, Jennifer Riley, Raphael Rahmani, Kevin D. Read, Malcolm J. McConville, Vicky M. Avery, Jonathan B. Baell (Lead / Corresponding author), Darren J. Creek (Lead / Corresponding author)

Research output: Contribution to journalArticle

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Abstract

Introduction: Trypanosoma brucei is the causative agent of human African trypanosomiasis, which is responsible for thousands of deaths every year. Current therapies are limited and there is an urgent need to develop new drugs. The anti-trypanosomal compound, 3-(oxazolo[4,5-b]pyridine-2-yl)anilide (OXPA), was initially identified in a phenotypic screen and subsequently optimized by structure–activity directed medicinal chemistry. It has been shown to be non-toxic and to be active against a number of trypanosomatid parasites. However, nothing is known about its mechanism of action. 

Objective: Here, we have utilized an untargeted metabolomics approach to investigate the biochemical effects and potential mode of action of this compound in T. brucei

Methods: Total metabolite extracts were analysed by HILIC-chromatography coupled to high resolution mass spectrometry. 

Results: Significant accumulation of ceramides was observed in OXPA-treated T. brucei. To further understand drug-induced changes in lipid metabolism, a lipidomics method was developed which enables the measurement of hundreds of lipids with high throughput and precision. The application of this LC–MS based approach to cultured bloodstream-form T. brucei putatively identified over 500 lipids in the parasite including glycerophospholipids, sphingolipids and fatty acyls, and confirmed the OXPA-induced accumulation of ceramides. Labelling with BODIPY-ceramide further confirmed the ceramide accumulation following drug treatment. 

Conclusion: These findings clearly demonstrate perturbation of ceramide metabolism by OXPA and indicate that the sphingolipid pathway is a promising drug target in T. brucei.

Original languageEnglish
Article number126
Number of pages14
JournalMetabolomics
Volume12
Issue number7
Early online date9 Jul 2016
DOIs
Publication statusPublished - Jul 2016

Fingerprint

Anilides
Trypanosoma brucei brucei
Sphingolipids
Metabolomics
Ceramides
Metabolism
Pharmaceutical Preparations
Parasites
Glycerophospholipids
African Trypanosomiasis
Lipids
Drug therapy
Pharmaceutical Chemistry
Metabolites
Chromatography
Lipid Metabolism
Labeling
Action Potentials
Mass spectrometry
Mass Spectrometry

Keywords

  • Human African trypanosomiasis
  • Lipidomics
  • Metabolomics
  • Sphingolipid metabolism
  • Trypanosoma brucei

Cite this

Stoessel, Daniel ; Nowell, Cameron J. ; Jones, Amy J. ; Ferrins, Lori ; Ellis, Katherine M. ; Riley, Jennifer ; Rahmani, Raphael ; Read, Kevin D. ; McConville, Malcolm J. ; Avery, Vicky M. ; Baell, Jonathan B. ; Creek, Darren J. / Metabolomics and lipidomics reveal perturbation of sphingolipid metabolism by a novel anti-trypanosomal 3-(oxazolo[4,5-b]pyridine-2-yl)anilide. In: Metabolomics. 2016 ; Vol. 12, No. 7.
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abstract = "Introduction: Trypanosoma brucei is the causative agent of human African trypanosomiasis, which is responsible for thousands of deaths every year. Current therapies are limited and there is an urgent need to develop new drugs. The anti-trypanosomal compound, 3-(oxazolo[4,5-b]pyridine-2-yl)anilide (OXPA), was initially identified in a phenotypic screen and subsequently optimized by structure–activity directed medicinal chemistry. It has been shown to be non-toxic and to be active against a number of trypanosomatid parasites. However, nothing is known about its mechanism of action. Objective: Here, we have utilized an untargeted metabolomics approach to investigate the biochemical effects and potential mode of action of this compound in T. brucei. Methods: Total metabolite extracts were analysed by HILIC-chromatography coupled to high resolution mass spectrometry. Results: Significant accumulation of ceramides was observed in OXPA-treated T. brucei. To further understand drug-induced changes in lipid metabolism, a lipidomics method was developed which enables the measurement of hundreds of lipids with high throughput and precision. The application of this LC–MS based approach to cultured bloodstream-form T. brucei putatively identified over 500 lipids in the parasite including glycerophospholipids, sphingolipids and fatty acyls, and confirmed the OXPA-induced accumulation of ceramides. Labelling with BODIPY-ceramide further confirmed the ceramide accumulation following drug treatment. Conclusion: These findings clearly demonstrate perturbation of ceramide metabolism by OXPA and indicate that the sphingolipid pathway is a promising drug target in T. brucei.",
keywords = "Human African trypanosomiasis, Lipidomics, Metabolomics, Sphingolipid metabolism, Trypanosoma brucei",
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Stoessel, D, Nowell, CJ, Jones, AJ, Ferrins, L, Ellis, KM, Riley, J, Rahmani, R, Read, KD, McConville, MJ, Avery, VM, Baell, JB & Creek, DJ 2016, 'Metabolomics and lipidomics reveal perturbation of sphingolipid metabolism by a novel anti-trypanosomal 3-(oxazolo[4,5-b]pyridine-2-yl)anilide', Metabolomics, vol. 12, no. 7, 126. https://doi.org/10.1007/s11306-016-1062-1

Metabolomics and lipidomics reveal perturbation of sphingolipid metabolism by a novel anti-trypanosomal 3-(oxazolo[4,5-b]pyridine-2-yl)anilide. / Stoessel, Daniel; Nowell, Cameron J.; Jones, Amy J.; Ferrins, Lori; Ellis, Katherine M.; Riley, Jennifer; Rahmani, Raphael; Read, Kevin D.; McConville, Malcolm J.; Avery, Vicky M.; Baell, Jonathan B. (Lead / Corresponding author); Creek, Darren J. (Lead / Corresponding author).

In: Metabolomics, Vol. 12, No. 7, 126, 07.2016.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Metabolomics and lipidomics reveal perturbation of sphingolipid metabolism by a novel anti-trypanosomal 3-(oxazolo[4,5-b]pyridine-2-yl)anilide

AU - Stoessel, Daniel

AU - Nowell, Cameron J.

AU - Jones, Amy J.

AU - Ferrins, Lori

AU - Ellis, Katherine M.

AU - Riley, Jennifer

AU - Rahmani, Raphael

AU - Read, Kevin D.

AU - McConville, Malcolm J.

AU - Avery, Vicky M.

AU - Baell, Jonathan B.

AU - Creek, Darren J.

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N2 - Introduction: Trypanosoma brucei is the causative agent of human African trypanosomiasis, which is responsible for thousands of deaths every year. Current therapies are limited and there is an urgent need to develop new drugs. The anti-trypanosomal compound, 3-(oxazolo[4,5-b]pyridine-2-yl)anilide (OXPA), was initially identified in a phenotypic screen and subsequently optimized by structure–activity directed medicinal chemistry. It has been shown to be non-toxic and to be active against a number of trypanosomatid parasites. However, nothing is known about its mechanism of action. Objective: Here, we have utilized an untargeted metabolomics approach to investigate the biochemical effects and potential mode of action of this compound in T. brucei. Methods: Total metabolite extracts were analysed by HILIC-chromatography coupled to high resolution mass spectrometry. Results: Significant accumulation of ceramides was observed in OXPA-treated T. brucei. To further understand drug-induced changes in lipid metabolism, a lipidomics method was developed which enables the measurement of hundreds of lipids with high throughput and precision. The application of this LC–MS based approach to cultured bloodstream-form T. brucei putatively identified over 500 lipids in the parasite including glycerophospholipids, sphingolipids and fatty acyls, and confirmed the OXPA-induced accumulation of ceramides. Labelling with BODIPY-ceramide further confirmed the ceramide accumulation following drug treatment. Conclusion: These findings clearly demonstrate perturbation of ceramide metabolism by OXPA and indicate that the sphingolipid pathway is a promising drug target in T. brucei.

AB - Introduction: Trypanosoma brucei is the causative agent of human African trypanosomiasis, which is responsible for thousands of deaths every year. Current therapies are limited and there is an urgent need to develop new drugs. The anti-trypanosomal compound, 3-(oxazolo[4,5-b]pyridine-2-yl)anilide (OXPA), was initially identified in a phenotypic screen and subsequently optimized by structure–activity directed medicinal chemistry. It has been shown to be non-toxic and to be active against a number of trypanosomatid parasites. However, nothing is known about its mechanism of action. Objective: Here, we have utilized an untargeted metabolomics approach to investigate the biochemical effects and potential mode of action of this compound in T. brucei. Methods: Total metabolite extracts were analysed by HILIC-chromatography coupled to high resolution mass spectrometry. Results: Significant accumulation of ceramides was observed in OXPA-treated T. brucei. To further understand drug-induced changes in lipid metabolism, a lipidomics method was developed which enables the measurement of hundreds of lipids with high throughput and precision. The application of this LC–MS based approach to cultured bloodstream-form T. brucei putatively identified over 500 lipids in the parasite including glycerophospholipids, sphingolipids and fatty acyls, and confirmed the OXPA-induced accumulation of ceramides. Labelling with BODIPY-ceramide further confirmed the ceramide accumulation following drug treatment. Conclusion: These findings clearly demonstrate perturbation of ceramide metabolism by OXPA and indicate that the sphingolipid pathway is a promising drug target in T. brucei.

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