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Phenotyping human blood plasma by 1H-NMR: a robust protocol based on metabolite spiking and its evaluation in breast cancer

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Abstract

This study reports an accurate assignment of the resonance signals present in 1H-NMR spectra of human blood plasma. Hereto, blood plasma was spiked with 32 different metabolites in relevant concentrations since reported chemical shift values show quite some variability depending on the biofluid under study and the applied experimental measuring conditions. The resulting information was used to rationally divide the 1H-NMR spectrum in 110 well-defined integration regions for application in metabolomics. A case–control dataset of 53 breast cancer patients and 52 controls was investigated in order to demonstrate the proof of principle. After removal of noisy variables, i.e. variables exceeding a premised threshold for the coefficient of variation, the groups could be discriminated by OPLS-DA multivariate statistics with a sensitivity and specificity of 83 and 94 %, respectively. In addition, the classification was validated in a small but independent cohort. The proposed methodology might pave the way towards a better understanding of disturbances in disease-related biochemical pathways and so, to the clinical relevance of study findings.

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Abbreviations

1H:

Proton

BMI:

Body mass index

CPMG:

Carr-Purcell-Meiboom-Gill

ChEBI:

Chemical Entities of Biological Interest

D2O:

Deuterium oxide

DCIS:

Ductal carcinoma in situ

DModX:

Distance to model

ER:

Estrogen receptor

GC:

Gas chromatography

IDA:

Invasive ductal adenocarcinoma

ILA:

Invasive lobular adenocarcinoma

IUPAC-IUB:

Nomenclature committee of the international union of biochemistry

LC:

Liquid chromatography

MHz:

Megahertz

MS:

Mass spectrometry

NMR:

Nuclear magnetic resonance

OPLS-DA:

Orthogonal partial least squares-discriminant analysis

PCA:

Principal component analysis

ppm:

Parts per million

PR:

Progesterone receptor

RF:

Radio frequency

S/N:

Signal-to-noise ratio

TSP:

Trimethylsilyl-2,2,3,3-tetradeuteropropionic acid

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Acknowledgments

This study is part of the Limburg Clinical Research Program UHasselt-ZOL-Jessa, supported by the foundation Limburg Sterk Merk, Hasselt University, Ziekenhuis Oost-Limburg and Jessa Hospital. We thank the Research Foundation Flanders (FWO-Vlaanderen) for their support via the MULTIMAR scientific research community project. We thank P. Degryse for his valuable contribution to the statistical discussions.

Ethical standards

All procedures followed were in accordance with the ethical standards of the responsible committee on human experimentation (institutional and national) and with the Helsinki Declaration of 1975, as revised in 2000 (5). Informed consent was obtained from all patients for being included in the study.

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Authors and Affiliations

  1. Faculty of Medicine and Life Sciences, Hasselt University, Martelarenlaan 42, 3500, Hasselt, Belgium

    Evelyne Louis, Liene Bervoets, Eric De Jonge, Liesbet Mesotten & Michiel Thomeer

  2. Institute for Materials Research, Hasselt University, Martelarenlaan 42, 3500, Hasselt, Belgium

    Gunter Reekmans & Peter Adriaensens

  3. Department of Gynaecology, Ziekenhuis Oost-Limburg, Schiepse Bos 6, 3600, Genk, Belgium

    Eric De Jonge

  4. Department of Nuclear Medicine, Ziekenhuis Oost-Limburg, Schiepse Bos 6, 3600, Genk, Belgium

    Liesbet Mesotten

  5. Department of Respiratory Medicine, Ziekenhuis Oost-Limburg, Schiepse Bos 6, 3600, Genk, Belgium

    Michiel Thomeer

Authors
  1. Evelyne Louis
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  2. Liene Bervoets
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  3. Gunter Reekmans
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  4. Eric De Jonge
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  5. Liesbet Mesotten
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  6. Michiel Thomeer
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  7. Peter Adriaensens
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Corresponding author

Correspondence to Peter Adriaensens.

Additional information

The authors Evelyne Louis and Liene Bervoets contributed equally to this work.

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Louis, E., Bervoets, L., Reekmans, G. et al. Phenotyping human blood plasma by 1H-NMR: a robust protocol based on metabolite spiking and its evaluation in breast cancer. Metabolomics 11, 225–236 (2015). https://doi.org/10.1007/s11306-014-0690-6

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  • DOI: https://doi.org/10.1007/s11306-014-0690-6

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