Dysregulation of the Tryptophan Pathway Evidences Gender Differences in COPD

Shama Naz, Maria Bhat, Sara Ståhl, Helena Forsslund, C Magnus Sköld, Åsa M Wheelock, Craig E Wheelock, Shama Naz, Maria Bhat, Sara Ståhl, Helena Forsslund, C Magnus Sköld, Åsa M Wheelock, Craig E Wheelock

Abstract

Increased activity of indoleamine 2,3-dioxygenase (IDO) and tryptophan hydroxylase (TPH) have been reported in individuals with chronic obstructive pulmonary disease (COPD). We therefore investigated the effect of gender stratification upon the observed levels of tryptophan metabolites in COPD. Tryptophan, serotonin, kynurenine, and kynurenic acid were quantified in serum of never-smokers (n = 39), smokers (n = 40), COPD smokers (n = 27), and COPD ex-smokers (n = 11) by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). The individual metabolite associations with lung function, blood, and bronchoalveolar lavage (BAL) immune-cell composition, as well as chemokine and cytokine levels, were investigated. Stratification by gender and smoking status revealed that the observed alterations in kynurenine and kynurenic acid, and to a lesser extent serotonin, were prominent in males, irrespective of COPD status (kynurenine p = 0.005, kynurenic acid p = 0.009, and serotonin p = 0.02). Inferred serum IDO activity and kynurenine levels decreased in smokers relative to never-smokers (p = 0.005 and p = 0.004, respectively). In contrast, inferred tryptophan hydroxylase (TPH) activity and serotonin levels showed an increase with smoking that reached significance with COPD (p = 0.01 and p = 0.01, respectively). Serum IDO activity correlated with blood CXC chemokine ligand 9 (CXCL9, p = 0.0009, r = 0.93) and chemokine (C-C motif) ligand 4 (CCL4.(p = 0.04, r = 0.73) in female COPD smokers. Conversely, serum serotonin levels correlated with BAL CD4+ T-cells (%) (p = 0.001, r = 0.92) and CD8+ T-cells (%) (p = 0.002, r = -0.90) in female COPD smokers, but not in male COPD smokers (p = 0.1, r = 0.46 and p = 0.1, r = -0.50, respectively). IDO- and TPH-mediated tryptophan metabolites showed gender-based associations in COPD, which were primarily driven by smoking status.

Keywords: COPD; IDO; gender; kynurenine; serotonin; smoking; tryptophan.

Conflict of interest statement

The authors declare no conflict of interest. Maria Bhat and Sara Ståhl were employed by AstraZeneca and may hold company shares. The involvement of AstraZeneca in this study included development of analysis method and analysis of samples.

Figures

Figure 1
Figure 1
Tryptophan pathway. Metabolites marked in bold were quantified by LC-MS/MS in the current study. IDO = indoleamine 2,3-dioxygenase, MAO = monoamine oxidase, TDO = tryptophan 2,3-dioxygenase, KAT = kynurenine aminotransferase, TPH = tryptophan hydroxylase.
Figure 2
Figure 2
Serum concentration of tryptophan pathway metabolites in smokers vs. COPD smokers, stratified by gender. (A) tryptophan, (B) kynurenine, (C) kynurenic acid, and (D) serotonin. Groups: Male smokers (n = 20), Male COPD (n = 14), Female smokers (n = 20), Female COPD (n = 12). Smokers: closed circles, COPD smokers: open circles, Male: blue, Female: orange. Significance was tested by applying a non-parametric Mann–Whitney test.
Figure 3
Figure 3
Serum concentration of tryptophan pathway metabolites in combined non-smokers vs. smokers irrespective of COPD status, stratified by gender. (A) tryptophan, (B) kynurenine, (C) kynurenic acid, and (D) serotonin. Groups: Male non-smokers (n = 24), Male smokers (n = 34), Female non-smokers (n = 25), Female smokers (n = 32). Smokers: closed circles, COPD smokers: open circles, male: blue, female: orange. Significance was tested by applying a non-parametric Mann–Whitney test. The corresponding p-values for the non-smoker vs. smoker combined gender comparisons were p = 0.6, p = 0.004, p = 0.008, and p = 0.009, respectively.
Figure 4
Figure 4
The level of serotonin in the current smoker COPD group stratified by chronic bronchitis (CB), emphysema (E) and absence of CB and E. Male: blue, Female: orange. Significance was tested using a non-parametric Mann–Whitney test.
Figure 5
Figure 5
(A) IDO activity was estimated by the ratio of kynurenine/tryptophan concentrations and (B) TPH activity was estimated by the ratio of serotonin/tryptophan concentrations. Never-smokers: closed circles, smokers: open circles, COPD smokers: closed triangles, COPD ex-smokers: open triangles. Male: blue, Female: orange. Significance was tested by applying a non-parametric Kruskal–Wallis one-way ANOVA and a non-parametric Mann–Whitney test.
Figure 6
Figure 6
Correlation of serum serotonin levels with T-cell populations from BAL cells in COPD smokers. (A) CD4/CD8 %, (B) CD4 %, (C) CD8 % (D) CD4+ FOXP3+ %, (E) CD8+ CD103+ CD69− CD27+ % and (F) CD8+ CD103+ CD69− CD27− %. Male: blue triangles, Female: orange triangles.

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