OP0304 METABOLOMICS PROFILING OF HUMAN SERUM FOR DISCOVERING BIOMARKERS TO DIAGNOSE PSORIATIC ARTHRITIS AND ANKYLOSING SPONDYLITIS WITH HIGH SPECIFICITY
Authors: W. Han, X. Wang, L. LI, S. Wichuk, E. Hutchings, R. Dadashova, J. Paschke, W. P. Maksymowych
Journal: Annals of the Rheumatic Diseases (2020)

Background: High-specificity biomarkers that can differentiate PsA and AS patients from healthy people, as well as RA patients, would be extremely helpful for early diagnosis and treatment of these two diseases. Based on the hypothesis that each disease state may cause specific changes to the metabolome, metabolomics is becoming a powerful tool for biomarker discovery. In this work, we applied a high-performance chemical isotope labeling (CIL) LC-MS platform to search for biomarker candidates of PsA and AS in human serum samples. Objectives: We aimed to identify metabolite biomarkers with high specificity for PsA and AS. Methods: Serum samples were collected from 331 subjects, including 100 healthy controls, 48 PsA patients, 52 AS patients and 131 RA patients. The average age of each group was: 52.6 (control), 50.7 (PsA), 51.8 (AS) and 53.1 (RA) years. After pre-treatment, each sample was incubated with 12 C-dansyl chloride, which can label the amine/phenol-containing metabolites. The reference sample for relative quantification was prepared by mixing all individual samples and then labeled by 13 C-dansyl chloride. With this normalization, the individual samples and the reference sample were mixed at an equal amount. Finally, we used an LC-QTOF-MS platform to analyze the mixtures and measure the 12 C/ 13 C peak pairs. Results: We detected 1,149 peak pairs commonly existing in the serum samples. Using our dansyl-library of 700 dansyl-labeled standards and a prediction library, which contains the predicted retention times and mass values of 3,431 dansylated human metabolites, we identified 134 and 141 peak pairs, respectively. The relative concentrations are calculated from the intensity ratios of 12 C/ 13 C peak pairs. We first visualized the entire amine/phenol-submetabolome for all phenotypes using the partial least squares discriminant analysis (PLS-DA). We found that the most significant between-group separation was between healthy controls and all the patients. No significant sex or age effect was observed. Furthermore, among the three diseases, PsA and AS samples were closely clustering, while the RA group was well separated from them. Therefore, we chose a two-step diagnosis approach that first differentiates PsA patients from controls/RA patients and then filters out the AS patients wrongly classified as PsA in the first step. The same strategy was conducted for AS. Stipulating a fold change larger than 1.5 with the false discovery rate lower than 5%, we found 74 metabolites having significantly higher or lower concentrations in the PsA group compared to the control or the RA group. We selected two of these significant metabolites to build a classification model based on the linear support vector machine (SVM) method, and the area-under-the-curve (AUC) value of the resulting receiver operating characteristic (ROC) curve was 0.929 (95% confidence interval: 0.899-0.956). Similarly, 37 metabolites could differentiate AS samples from RAs and controls. A proposed diagnostic panel containing four metabolites demonstrated an AUC value of 0.890 (0.843-0.934). For the last step, distinguishing between PsA and AS, there were 15 significantly increased metabolites and 9 lowered ones. The biomarker panel consisting of the top three metabolites also achieved good discriminatory power with AUC = 0.827 (0.717-0.919). Conclusion: Isotope-labeling-LC-MS-based metabolomics has revealed biomarker candidates that can specifically differentiate PsA or AS patients from control populations. Disclosure of Interests: Wei Han: None declared, Xiaohang Wang: None declared, Liang Li: None declared, Stephanie Wichuk: None declared, Edna Hutchings: None declared, Rana Dadashova: None declared, Joel Paschke: None declared, Walter P Maksymowych Grant/research support from: Received research and/or educational grants from Abbvie, Novartis, Pfizer, UCB, Consultant of: WPM is Chief Medical Officer of CARE Arthritis Limited, has received consultant/participated in advisory boards for Abbvie, Boehringer, Celgene, Eli-Lilly, Galapagos, Gilead, Janssen, Novartis, Pfizer, UCB, Speakers bureau: Received speaker fees from Abbvie, Janssen, Novartis, Pfizer, UCB.