Metabolomics analysis identifies sex-associated metabotypes of oxidative stress and the autotaxin-lysoPA axis in COPD

Shama Naz, Johan Kolmert, Mingxing Yang, Stacey N Reinke, Muhammad Anas Kamleh, Stuart Snowden, Tina Heyder, Bettina Levänen, David J Erle, C Magnus Sköld, Åsa M Wheelock, Craig E Wheelock, Shama Naz, Johan Kolmert, Mingxing Yang, Stacey N Reinke, Muhammad Anas Kamleh, Stuart Snowden, Tina Heyder, Bettina Levänen, David J Erle, C Magnus Sköld, Åsa M Wheelock, Craig E Wheelock

Abstract

Chronic obstructive pulmonary disease (COPD) is a heterogeneous disease and a leading cause of mortality and morbidity worldwide. The aim of this study was to investigate the sex dependency of circulating metabolic profiles in COPD.Serum from healthy never-smokers (healthy), smokers with normal lung function (smokers), and smokers with COPD (COPD; Global Initiative for Chronic Obstructive Lung Disease stages I-II/A-B) from the Karolinska COSMIC cohort (n=116) was analysed using our nontargeted liquid chromatography-high resolution mass spectrometry metabolomics platform.Pathway analyses revealed that several altered metabolites are involved in oxidative stress. Supervised multivariate modelling showed significant classification of smokers from COPD (p=2.8×10-7). Sex stratification indicated that the separation was driven by females (p=2.4×10-7) relative to males (p=4.0×10-4). Significantly altered metabolites were confirmed quantitatively using targeted metabolomics. Multivariate modelling of targeted metabolomics data confirmed enhanced metabolic dysregulation in females with COPD (p=3.0×10-3) relative to males (p=0.10). The autotaxin products lysoPA (16:0) and lysoPA (18:2) correlated with lung function (forced expiratory volume in 1 s) in males with COPD (r=0.86; p<0.0001), but not females (r=0.44; p=0.15), potentially related to observed dysregulation of the miR-29 family in the lung.These findings highlight the role of oxidative stress in COPD, and suggest that sex-enhanced dysregulation in oxidative stress, and potentially the autotaxin-lysoPA axis, are associated with disease mechanisms and/or prevalence.

Conflict of interest statement

Conflict of interest: Disclosures can be found alongside this article at erj.ersjournals.com

Copyright ©ERS 2017.

Figures

FIGURE 1
FIGURE 1
Optimised orthogonal projections to latent structures discriminant analysis multivariate models using nontargeted metabolomics data. a) i) Scores plot of male smokers versus males with chronic obstructive pulmonary disease (COPD) (R2Y=0.49, Q2=0.38, p=4.0×10−4); ii) loadings of confirmed metabolites that were the most prominent for driving the separation between male smokers versus males with COPD. b) i) Scores plot of female smokers versus females with COPD (R2Y=0.73, Q2=0.65, p=2.4×10−7); ii) loadings of confirmed metabolites that were the most prominent for driving the separation of female smokers versus females with COPD. For ease of display, parts a ii) and b ii) exclude metabolites whose standard error crossed the x-axis. The complete list of loadings is shown in online supplementary figure E8. LysoPA: lyso-phosphatidic acid; HETE: hydroxyeicosatetraenoic acid; HDoHE: hydroxydocosahexaenoic acid; EpETrE: epoxyeicosatrienoic acid; HEDE: hydroxyeicosadienoic acid; AEA: N-arachidonoylethanolamine; OEA: N-oleoylethanolamine; EpODE: epoxyoctadecadienoic acid; PEA: N-palmitoylethanolamide; Asp-Leu: aspartic acid-leucine.
FIGURE 2
FIGURE 2
The lyso-phosphatidic acid (lysoPA)–autotaxin axis was attenuated in males with chronic obstructive pulmonary disease (COPD). a) Serum lysoPA (16:0) levels in smokers versus COPD; (b) serum lysoPA (18:2) levels in smokers versus COPD; (c) lysoPA (16:0) and lysoPA (18:2) metabolites correlated with lung function (forced expiratory volume in 1 s (FEV1)) in male COPD patients (r=0.84, p<0.0001). No correlation was observed in the corresponding female COPD population (r=0.44, p=0.15); (d) levels of miR-29b in bronchoalveolar lavage (BAL) cells from male and female smokers and COPD patients. Values for the other members of the miR-29 family are shown in online supplementary figure E6. LysoPA data are from the nontargeted metabolomics platform and are presented as log2 of arbitrary units (AU). RFU: relative fluorescence units; LLOQ: lower limit of quantification.
FIGURE 3
FIGURE 3
β-oxidation-related metabolite ratio of carnitine with acylcarnitines in relation to sex and disease status for smoking subjects. a) Ratio of carnitine with sum of the medium-chain carnitines; (b) ratio of carnitine with sum of the long-chain carnitines. Subjects are divided into smokers with normal lung function and smokers with chronic obstructive pulmonary disease (COPD). Significance was calculated using the nonparametric Mann–Whitney test. Data are from the targeted metabolomics method (Biocrates).
FIGURE 4
FIGURE 4
Serum levels of analytes involved in the arginine/nitric oxide pathway. a) Ratio of acetyl-ornithine to ornithine; (b) ratio of total arginine to the inferred activity of the nitric oxide synthase (NOS) enzyme expressed as arginine/(ornithine+citrulline); (c) ratio of endogenous NOS inhibitors (sum of asymmetric and symmetric dimethylarginine (ADMA + SDMA)) with arginine; and (d) concentration of the endogenous NOS inhibitor ADMA. Significance was calculated using the nonparametric Mann–Whitney test. Subjects are divided into smokers with normal lung function and smokers with chronic obstructive pulmonary disease (COPD). Data are from the targeted metabolomics method (Biocrates).
FIGURE 5
FIGURE 5
Representative pathway outline for the altered metabolites involved in oxidative stress metabolism in chronic obstructive pulmonary disease (COPD). (a) Fatty acid β-oxidation pathway; (b) purine degradation pathway; (c) Land's cycle/phospholipid metabolism. TCA: tricarboxylic acid; NADH: nicotinamide adenine dinucleotide; FADH2: flavin adenine dinucleotide; IMP: inosine monophosphate; NAPE-PLD: N-acyl phosphatidylethanolamine phospholipase D; lysoPA: lysophosphatidic acid.

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