Index markers of chronic fatigue syndrome with dysfunction of TCA and urea cycles

Emi Yamano, Masahiro Sugimoto, Akiyoshi Hirayama, Satoshi Kume, Masanori Yamato, Guanghua Jin, Seiki Tajima, Nobuhito Goda, Kazuhiro Iwai, Sanae Fukuda, Kouzi Yamaguti, Hirohiko Kuratsune, Tomoyoshi Soga, Yasuyoshi Watanabe, Yosky Kataoka, Emi Yamano, Masahiro Sugimoto, Akiyoshi Hirayama, Satoshi Kume, Masanori Yamato, Guanghua Jin, Seiki Tajima, Nobuhito Goda, Kazuhiro Iwai, Sanae Fukuda, Kouzi Yamaguti, Hirohiko Kuratsune, Tomoyoshi Soga, Yasuyoshi Watanabe, Yosky Kataoka

Abstract

Chronic fatigue syndrome (CFS) is a persistent and unexplained pathological state characterized by exertional and severely debilitating fatigue, with/without infectious or neuropsychiatric symptoms, lasting at least 6 consecutive months. Its pathogenesis remains incompletely understood. Here, we performed comprehensive metabolomic analyses of 133 plasma samples obtained from CFS patients and healthy controls to establish an objective diagnosis of CFS. CFS patients exhibited significant differences in intermediate metabolite concentrations in the tricarboxylic acid (TCA) and urea cycles. The combination of ornithine/citrulline and pyruvate/isocitrate ratios discriminated CFS patients from healthy controls, yielding area under the receiver operating characteristic curve values of 0.801 (95% confidential interval [CI]: 0.711-0.890, P < 0.0001) and 0.750 (95% CI: 0.584-0.916, P = 0.0069) for training (n = 93) and validation (n = 40) datasets, respectively. These findings provide compelling evidence that a clinical diagnostic tool could be developed for CFS based on the ratios of metabolites in plasma.

Figures

Figure 1. Metabolic pathway map of quantified…
Figure 1. Metabolic pathway map of quantified metabolite concentrations, including for glycolysis, the tricarboxylic acid cycle, the urea cycle and glutamine metabolism, in chronic fatigue syndrome (CFS) patients and healthy controls (HCs).
Box-and-whisker plots of the concentrations of metabolites involved in energy metabolism in the plasma of HCs and CFS patients. The coloured plots denote HCs (green) and CFS patients (red). The horizontal lines indicate the minimum, maximum, median, and first and third quartile. #P < 0.10; *P < 0.05; **P < 0.01 (Mann–Whitney U-test).
Figure 2. Box-and-whisker plots of ratios of…
Figure 2. Box-and-whisker plots of ratios of pyruvate/isocitrate and ornithine/citrulline in training and validation datasets.
The horizontal lines indicate the minimum, maximum, median, and first and third quartile. *P < 0.05; ***P < 0.001; ****P < 0.0001 (Mann–Whitney U-test).
Figure 3. Overall flow of the development…
Figure 3. Overall flow of the development and validation of the multiple logistic regression model and results predicted by this model.
(a) The training data were used for development and internal validation of the model, starting from support vector machine-feature selection, feature selection based on metabolic pathways, model development and internal validation. The model predicted independent validation datasets that were not used for model training. (b) Box-and-whisker plots of the probability of chronic fatigue syndrome yielded by the developed model. Horizontal lines indicate the minimum, maximum, median and first and third quartile. (c) Receiver operating characteristic curves for training and validation datasets.

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