Discovery of Potential Serum Protein Biomarkers in Ankylosing Spondylitis Using TMT-Based Quantitative Proteomics
23 Pages Posted: 27 Jul 2019More...
Background: Ankylosing Spondylitis (AS) is a systemic, chronic and inflammatory rheumatic disease, which affected 0.2% of the population. Current diagnostic criteria relys on a composite of clinical and rediological changes, with an average of 14 years from AS symptoms onset to diagnosis. Here, we aimed to discover a panel of potential biomarkers for AS diagnosis.
Methods: A cohort of 403 patients including 153 AS and 250 healthy subjects from 2 independent centers was studied. In the discovery set 1, TMT-based quantitative proteomics method was applied to identify serum proteins. In the discovery set 2, ELISA method was further performed to quantify and validate the levels of differential proteins from proteomics. An array of potential biomarkers were screened by the area under the receiver operating characteristic (ROC) curve. Finally, an accurate and reliable biomarkers panel was evaluated by the prediction accuracies in internal and external validation sets.
Findings: A total of 762 proteins were identified from 15 pooled AS and 60 pooled healthy serum samples. Among them, 46 up-regulated and 59 down-regulated proteins were identified in AS compared to healthy subjects. The AS responsive proteins including CRP, SAA1, ORM2, FG-γ and THBS1 were validated successfully by ELISA using sera of 36 AS and 36 healthy subjects. The area under the ROC curve of the CRP and SAA1 combination was high, near by 0.9. The sensitivity was 0.970, specificity was 0.805 at 95% confidence interval from 0.811 to 0.977. The predictive values reached 92.00% in the internal validation set (62 AS vs 114 healthy subjects) and 97.50% in the external validation phase (40 AS vs 40 healthy subjects).
Interpretation: A panel of two acute phase proteins (CRP and SAA1) as potential biomarkers could be useful for complementary diagnosis of AS.
Funding: This work is financially supported by the National Natural Science Foundation of China,
the Natural Science Foundation of Jiangsu province, Jiangsu Provincial Medical Youth Talent (No.
QNRC2016642), the Young Elite Scientists Sponsorship Program by CAST (No. QNRC2-B04), Science and technology projects of Jiangsu Province Hospital of TCM (No. Y18010) and the top
six talent project of Jiangsu province 2016 (WSN-051).
Declaration of Interest: There is no any conflict of interest with regard to our work.
Ethical Approval: All procedures were approved by the medical ethics committee of the affiliated Hospital of Nanjing University of Chinese Medicine and followed the tenets of the Declaration of Helsinki. The experimental protocol was reviewed and approved by the Institutional Review Board of the affiliated Hospital of Nanjing University of Chinese Medicine (2018NL-106-02).
Keywords: Biomarker, Ankylosing Spondylitis, CRP, SAA1, Serum proteomics
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