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Serum amino acid metabolic profiles of ankylosing spondylitis by targeted metabolomics analysis

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Abstract

Background

Ankylosing spondylitis (AS) is a common chronic inflammatory arthritis, causing lasting back pain with progressive loss of spinal mobility. However, the exact pathogenesis of AS remains unclear. We aim to use the metabolomics analysis to characterize the metabolic profile of AS, in order to better understand the pathogenesis of AS and monitor disease activity and progression.

Methods

The ultra-high performance liquid chromatography-triple quadrupole mass spectrometry (UPLC-TQ-MS) was used for investigating the serum amino acid metabolomic profiling of 30 AS patients, in comparison with 32 rheumatoid arthritis (RA) patients and 30 healthy controls, combined with multivariate statistical analysis. Metabolite association analysis with disease activity was performed using generalized linear regression. The metabolic pathway analysis for the important metabolites was performed using MetPA and the metabolic network was constructed.

Results

A total of 29 amino acids and biogenic amines were detected in all participants by UPLC-TQ-MS. It showed significant amino acid differences between the AS/RA patients and control subjects. Additionally, 4-hydroxy-L-proline, alanine, γ-aminobutyric acid, glutamine, and taurine were identified as candidate markers shared by AS/RA groups. Specifically, lysine, proline, serine, and alanine were found correlated with disease activity of AS. Furthermore, the most significant metabolic pathway identified were alanine, aspartate, and glutamate metabolism, arginine and proline metabolism, aminoacyl tRNA biosynthesis and glycine, serine, and threonine metabolism.

Conclusions

These preliminary results demonstrate that UPLC-TQ-MS analysis method is a powerful tool to identify metabolite profiles of AS. Research in identified disease activity–associated metabolites and biological pathways may provide assistance for clinical diagnosis and pathological mechanism of AS.

Key Points

• There are perturbations of serum amino acid metabolism in AS, compared with RA and healthy controls, determined by UPLC-TQ-MS.

• Metabolomics pathway is used to analysis for the differential metabolites of AS.

• The altered serum amino acid could monitor disease activity of AS.

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Abbreviations

AS:

Ankylosing spondylitis

RA:

Rheumatoid arthritis

UPLC-TQ-MS:

Ultra-high performance liquid chromatography-triple quadrupole mass spectrometry

ESI:

Electrospray ionization

PCA:

Principal component analysis

PLS-DA:

Partial least squares discriminant analysis

VIP:

Variable importance for project

KEGG:

The Kyoto Encyclopedia of Genes and Genomes

CRP:

C reaction protein

BCAAs:

Branched-chain amino acids

AAs:

Amino acids

HC:

healthy control

QC:

control sample

VIF:

Variance inflation factor

BMI:

Body mass index

ESR:

Erythrocyte sedimentation rate

BASDAI:

Bath Ankylosing Spondylitis Disease Activity Index

BASFI:

Bath Ankylosing Spondylitis Functional Index

ASDAS-CRP:

Ankylosing Spondylitis Disease Activity Score

mSASSS:

modified Stoke Ankylosing Spondylitis Spinal Score

ACPA:

anti-citrullinated protein/peptide antibodies

DAS28:

Disease activity score 28

RF:

Rheumatoid factor

ANA:

Antinuclear antibodies

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Funding

This work was supported by the Harbin medical university scientific research innovation fund (No. 2017LCZX98) and the Foundation of Heilongjiang Provincial Health Bureau (No. 2017-146), and Heilongjiang province postdoctoral fund (LBH-Z17142).

Author information

Author notes

  1. Yong Zhou and Xi Zhang contributed equally to this work.

Authors and Affiliations

  1. Department of Orthopaedic Surgery, The Fourth Affiliated Hospital of Harbin Medical University, Harbin, People’s Republic of China

    Yong Zhou, Xi Zhang, Rui Chen, Yishu Liu, Xuefeng Liu, Ming Gao, Chundong Yang & Daifeng Lu

  2. Department of Parasitology, Harbin Medical University, Harbin, People’s Republic of China

    Su Han, Beibei Sun & Hao Chen

Authors
  1. Yong Zhou
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  2. Xi Zhang
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Contributions

R.C., X.Z., and Y. Z. conceived and designed the experiments; R.C., Y.Z., X.Z., and S.H. performed the experiments; R.C. and S.H. analyzed the data; B.S., H.C., Y.L., X.L., M.G., C.Y., and D.L. contributed reagents/materials/analysis tools; R.C. and S.H. wrote the paper.

Corresponding author

Correspondence to Rui Chen.

Ethics declarations

The study was approved by the Ethics Committee of Harbin Medical University and conducted in accordance with the tenets of the Declaration of Helsinki. Written informed consent was approved by the Ethics Committee of Harbin Medical University and collected from each participant included in the study.

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Supplementary date (Fig. S1-S2 and Table S1-S16) associated with this article can be found in the online version.

ESM 1

(DOC 411 kb)

Fig. S1

PCA scores plot showing clustering of quality control (QC) samples. Red, QC; Blue, Healthy control. (PNG 157 kb)

High resolution image (TIF 228 kb)

Fig. S2

Mapping the rheumatoid arthritis-related metabolites in metabolic pathways. The metabolites marked in red indicate significantly increased in rheumatoid arthritis, and green indicates decreased, blue indicates unchanged and gray indicates un-investigated. (PNG 102 kb)

High resolution image (TIF 390 kb)

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Zhou, Y., Zhang, X., Chen, R. et al. Serum amino acid metabolic profiles of ankylosing spondylitis by targeted metabolomics analysis. Clin Rheumatol 39, 2325–2336 (2020). https://doi.org/10.1007/s10067-020-04974-z

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