Metabonomics of human fecal extracts characterize ulcerative colitis, Crohn's disease and healthy individuals

Jacob Tveiten Bjerrum, Yulan Wang, Fuhua Hao, Mehmet Coskun, Christian Ludwig, Ulrich Günther, Ole Haagen Nielsen, Jacob Tveiten Bjerrum, Yulan Wang, Fuhua Hao, Mehmet Coskun, Christian Ludwig, Ulrich Günther, Ole Haagen Nielsen

Abstract

This study employs spectroscopy-based metabolic profiling of fecal extracts from healthy subjects and patients with active or inactive ulcerative colitis (UC) and Crohn's disease (CD) to substantiate the potential use of spectroscopy as a non-invasive diagnostic tool and to characterize the fecal metabolome in inflammatory bowel disease (IBD). Stool samples from 113 individuals (UC 48, CD 44, controls 21) were analyzed by 1H nuclear magnetic resonance (NMR) spectroscopy (Bruker 600 MHz, Bruker BioSpin, Rheinstetten, Germany). Data were analyzed with principal component analysis and orthogonal-projection to latent structure-discriminant analysis using SIMCA-P + 12 and MATLAB. Significant differences were found in the metabolic profiles making it possible to differentiate between active IBD and controls and between UC and CD. The metabolites holding differential power primarily belonged to a range of amino acids, microbiota-related short chain fatty acids, and lactate suggestive of an inflammation-driven malabsorption and dysbiosis of the normal bacterial ecology. However, removal of patients with intestinal surgery and anti-TNF-α antibody treatment eliminated the discriminative power regarding UC versus CD. This study consequently demonstrates that 1H NMR spectroscopy of fecal extracts is a potential non-invasive diagnostic tool and able to characterize the inflammation-driven changes in the metabolic profiles related to malabsorption and dysbiosis. Intestinal surgery and medication are to be accounted for in future studies, as it seems to be factors of importance in the discriminative process.

Keywords: Diagnostic tool; Dysbiosis; Inflammatory bowel disease; Metabolomics; NMR spectroscopy.

Figures

Fig. 1
Fig. 1
OPLS-DA score plots. The score plots (a, b, c, and d) are based on the four valid models containing all patients and display the 1st PLS component and one orthogonal component for each model. A two-way separation of the fecal samples is demonstrated in all 4 plots. Blue diamonds control; empty triangles active CD; red dots inactive CD; empty circles active UC; purple stars: inactive UC. The corresponding back-scaled loading plots reflect the class differences in the NMR spectra. Upright peaks indicate a relatively increased intensity of metabolites, and downright peaks a decreased intensity of metabolites. The colors shown on the plot are associated with the significance of metabolites in separating the samples as shown on the right hand side of the plot, where the color-scaling map is given together with the respective correlation coefficients. In accordance with the sample number in each group and a significance level of p < 0.05, the metabolites are significant at correlation coefficient values above a 0.55, b 0.43, c 0.44, and d 0.38, respectively. CD Crohn’s disease; UC ulcerative colitis; ala alanine; asp aspartate; buty butyrate; glu glutamate; gly glycine; ileu isoleucine; lac lactate; leu leucine; lys lysine; phe phenylalanine; prop proprionate; tyr tyrosine; val valine (Color figure online)
Fig. 2
Fig. 2
OPLS-DA score plots of patients without intestinal surgery. The score plots (a and b) are based on the two valid models containing only patients without intestinal surgery and display the 1st PLS component and one orthogonal component for each model. A two-way separation of the fecal samples is demonstrated in both plots. Blue squares inactive UC; purple stars active UC; green triangles controls. The corresponding back-scaled loading plots reflect the class differences in the NMR spectra. Upright peaks indicate a relatively increased intensity of metabolites, and downright peaks a decreased intensity of metabolites. The colors shown on the plot are associated with the significance of metabolites in separating the samples as shown on the right hand side of the plot, where the color-scaling map is given together with the respective correlation coefficients in accordance with the sample number in each group and a significance level of p < 0.05, the metabolites are significant at correlation coefficient values above a 0.44 and b 0.44, respectively. UC ulcerative colitis; ala alanine; buty butyrate; ileu isoleucine; lac lactate; leu leucine; lys lysine; prop proprionate; tau taurine; val valine (Color figure online)

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