Metabolomics

, 12:126 | Cite as

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
  • Darren J. Creek
Original Article
Part of the following topical collections:
  1. Recent advances in Pharmacometabolomics: enabling tools for precision medicine

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 Trypanosoma brucei Metabolomics Lipidomics Sphingolipid metabolism 

Notes

Funding

DJC acknowledges support from a NHMRC training fellowship. LF acknowledges an Australian Postgraduate Award. MJM is an NHMRC Principal Research Fellow. Financial support was received from NHMRC project Grants APP1025581 and APP1067728.

Compliance with ethical standards

Conflict of interests

All authors declare that they have no conflict of interest.

Human and animal rights

No human participants or animals were involved in this study.

Supplementary material

11306_2016_1062_MOESM1_ESM.xlsb (30.8 mb)
S1IDEOM metabolite list and metadata from HILIC metabolomics study (XLSB 31541 kb)
11306_2016_1062_MOESM2_ESM.xlsb (22.9 mb)
S2IDEOM metabolite list and metadata from lipidomics study (XLSB 23466 kb)
11306_2016_1062_MOESM3_ESM.pdf (376 kb)
S3MSMS spectra of significant ceramides and acetylcarnitine (PDF 375 kb)
11306_2016_1062_MOESM4_ESM.pdf (39 kb)
S4Results from IC50 analysis in presence of Carnitine (PDF 38 kb)

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Daniel Stoessel
    • 1
    • 2
  • Cameron J. Nowell
    • 3
  • Amy J. Jones
    • 4
  • Lori Ferrins
    • 5
  • Katherine M. Ellis
    • 1
  • Jennifer Riley
    • 6
  • Raphael Rahmani
    • 5
  • Kevin D. Read
    • 6
  • Malcolm J. McConville
    • 7
  • Vicky M. Avery
    • 4
  • Jonathan B. Baell
    • 5
  • Darren J. Creek
    • 1
  1. 1.Drug Delivery, Disposition and Dynamics, Monash Institute of Pharmaceutical SciencesMonash UniversityParkvilleAustralia
  2. 2.Department of BiotechnologyBeuth University of Applied SciencesBerlinGermany
  3. 3.Drug Discovery Biology, Monash Institute of Pharmaceutical SciencesMonash UniversityParkvilleAustralia
  4. 4.Discovery Biology, Eskitis Institute for Drug DiscoveryGriffith UniversityNathanAustralia
  5. 5.Department of Medicinal Chemistry, Monash Institute of Pharmaceutical SciencesMonash UniversityParkvilleAustralia
  6. 6.Drug Discovery Unit, Division of Biological Chemistry and Drug Discovery, College of Life SciencesUniversity of DundeeDundeeUK
  7. 7.Department of Biochemistry and Molecular Biology, Bio21 Molecular Science and Biotechnology InstituteUniversity of MelbourneParkvilleAustralia

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