Plasma metabolomics in human pulmonary tuberculosis disease: a pilot study - PubMed (original) (raw)

. 2014 Oct 15;9(10):e108854.

doi: 10.1371/journal.pone.0108854. eCollection 2014.

Dean P Jones 1, Nestan Tukvadze 2, Karan Uppal 3, Eka Sanikidze 2, Maia Kipiani 2, ViLinh T Tran 3, Gautam Hebbar 3, Douglas I Walker 4, Russell R Kempker 3, Shaheen S Kurani 3, Romain A Colas 5, Jesmond Dalli 5, Vin Tangpricha 6, Charles N Serhan 5, Henry M Blumberg 7, Thomas R Ziegler 1

Affiliations

Plasma metabolomics in human pulmonary tuberculosis disease: a pilot study

Jennifer K Frediani et al. PLoS One. 2014.

Abstract

We aimed to characterize metabolites during tuberculosis (TB) disease and identify new pathophysiologic pathways involved in infection as well as biomarkers of TB onset, progression and resolution. Such data may inform development of new anti-tuberculosis drugs. Plasma samples from adults with newly diagnosed pulmonary TB disease and their matched, asymptomatic, sputum culture-negative household contacts were analyzed using liquid chromatography high-resolution mass spectrometry (LC-MS) to identify metabolites. Statistical and bioinformatics methods were used to select accurate mass/charge (m/z) ions that were significantly different between the two groups at a false discovery rate (FDR) of q<0.05. Two-way hierarchical cluster analysis (HCA) was used to identify clusters of ions contributing to separation of cases and controls, and metabolomics databases were used to match these ions to known metabolites. Identity of specific D-series resolvins, glutamate and Mycobacterium tuberculosis (Mtb)-derived trehalose-6-mycolate was confirmed using LC-MS/MS analysis. Over 23,000 metabolites were detected in untargeted metabolomic analysis and 61 metabolites were significantly different between the two groups. HCA revealed 8 metabolite clusters containing metabolites largely upregulated in patients with TB disease, including anti-TB drugs, glutamate, choline derivatives, Mycobacterium tuberculosis-derived cell wall glycolipids (trehalose-6-mycolate and phosphatidylinositol) and pro-resolving lipid mediators of inflammation, known to stimulate resolution, efferocytosis and microbial killing. The resolvins were confirmed to be RvD1, aspirin-triggered RvD1, and RvD2. This study shows that high-resolution metabolomic analysis can differentiate patients with active TB disease from their asymptomatic household contacts. Specific metabolites upregulated in the plasma of patients with active TB disease, including Mtb-derived glycolipids and resolvins, have potential as biomarkers and may reveal pathways involved in TB disease pathogenesis and resolution.

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Conflict of interest statement

Competing Interests: The authors have declared that no competing interests exist.

Figures

Figure 1

Figure 1. Plasma metabolome-wide association study (MWAS) of pulmonary tuberculosis (TB) disease in adults.

(A) The Manhattan plot depicts the -log P analysis of 23,241 metabolites comparing 17 adults with newly diagnosed pulmonary TB disease and 17 asymptomatic adult household contacts who were sputum smear and culture negative for Mtb. The x-axis represents the m/z of the metabolites, ordered in increasing value from left (85) to right (2000). A total of 61 metabolites significantly differed [false discovery rate (FDR) q = 0.05] between the two groups (metabolites depicted above the horizontal dashed blue line). Metabolites above the horizontal dashed red line (n = 122) distinguished the TB disease and household contact cohorts at FDR q = 0.10, while metabolites above the dashed green line (n = 711) distinguished the two groups at FDR q = 0.20. (B) Box-and-whisker plots of log2 intensities comparing individuals with TB disease and household contacts for selected metabolites, with m/z and putative metabolite identification from Metlin and KEGG (left to right upper panel: glutamate, D-series resolvin, and trehalose-6-mycolate, respectively; left to right lower panel: phosphatidylinositol, and two unidentified metabolites that did not match to known metabolites in the databases, respectively).

Figure 2

Figure 2. Significant metabolites that distinguish TB patients from household contacts.

(A) Two-way hierarchical cluster analysis (HCA) using C18 chromatography shows 8 clusters of metabolites from human plasma and illustrates the patterns distinguishing those with active TB from household contacts without evidence of TB disease. The 17 subjects with TB disease (TB; shown in green) and the 17 household contacts (HC; shown in red) are shown along the x-axis. (B) Pie chart depicts chemical classes of the 61 significant metabolites from panel 2A according to high-resolution matches to metabolite databases .

Figure 3

Figure 3. MS/MS fragmentation spectra show positive identification of resolvin D1 (RvD1), resolvin D2 (RvD2), and aspirin-triggered resolvin D1 (AT-RvD1) in plasma from subjects with TB disease.

Metabololipidomics analytical methods that incorporated high-resolution liquid chromatography coupled with tandem mass spectroscopy (LC-MS/MS, ABI 5500, see methods) were used to verify these DHA-derived specialized pro-resolving lipid mediators , .

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