A Serum Biomarker Panel of exomiR-451a, exomiR-25-3p and Soluble TWEAK for Early Diagnosis of Rheumatoid Arthritis

Abstract

BackgroundThe etiology of rheumatoid arthritis (RA) remains poorly understood. Early and accurate diagnosis still difficult to achieve. Inflammatory related molecules released into the circulation such cytokines and exosome-derived microRNAs (exomiRNAs) could be good candidates for early diagnosis of autoimmune diseases. We sought to discover a serum biomarker panel for the early detection of RA based on exomiRNAs and inflammatory markers.MethodsA 179 miRNAs-microarray panel was analyzed in a pilot study (4 early RA and 4 controls). Validation of deregulated exomiRNAs was performed in a larger cohort (24 patients with early RA and 24 controls). miRNet software was used to predict exomiRNA gene-targets interactions. Potentially altered pathways were analyzed by Reactome pathway database search. STRING database was used to predict protein-protein interaction networks. Enzyme-linked immunosorbent assay was used to measure serum levels of sTWEAK and sCD163. Signature biomarker candidates were statistical analyzed.ResultsWe detected 11 differentially expressed exomiRNAs in early RA pilot study. Validation analysis revealed that 6/11 exomiRNAs showed strong agreement with the pilot microarray data (exomiR-144-3p, -25-3p, -15a-5p, -451a, -107 and -185-5p). sTWEAK and sCD163 biomarkers were significantly elevated in the serum of patients with early RA. Receiver operating characteristic (ROC) analysis showed that the best panel to diagnose early RA contained exomiR-451a, exomiR-25-3p and sTWEAK, and could correctly classify 95.6% of patients, with an area under the ROC curve of 0.983 and with 100% specificity and 85.7% sensitivity. The YWHAB gene was identified as a common target of the putative miRNA-regulated pathways.ConclusionA novel serum biomarker panel composed of exomiR-451a, exomiR-25-3p and serum levels of sTWEAK may have use in the early clinical diagnosis of RA. A new predicted exomiRNA-target gene YHWAB has been identified and may have a relevant role in the development of RA.

The roles of extracellular vesicles in the immune system

“… Certain EV-associated microRNAs (miR-16, miR-302d-3p, miR-378e, miR-570-3p, miR-574-5p, miR-579 and miR-25-3p) are candidate biomarkers for type 1 diabetes 155 . Serum EV-associated miR-451a and miR-25-3p and soluble TWEAK (also known as TNFSF12) are candidate biomarkers in early rheumatoid arthritis 156 . …”

“…Serum EV-associated miR-451a and miR-25-3p and soluble TWEAK (also known as TNFSF12) are candidate biomarkers in early rheumatoid arthritis 156 .…”

Comprehensive overview of microRNA function in rheumatoid arthritis

“…245,246 Focusing on the unique miRNA profiles in serum exosomes from patients with early-stage untreated RA, Rodriguez-Muguruza et al developed a biomarker panel comprising serum exo-miR-25-3p, exo-miR-451a, and soluble TNF-like weak inducer of apoptosis (TWEAK), an inflammatory cytokine involved in the pathogenesis of RA, that may exhibit superior diagnostic utility over ACPA positivity. 247 More reproducible studies with expanded sample sizes and designs optimized to assess panel specificity are needed in the future. Two important confounding factors affecting the utility of exosomal miRNAs as biomarkers for RA are the diverse approaches used to isolate miRNAs from body fluids and the difficulty in identifying their cellular origin.…”

Knowledge Mapping of Exosomes in Autoimmune Diseases: A Bibliometric Analysis (2002–2021)

“…Lakhter A et al found that before the onset of T1D, the level of miR-21-5p in serum exosomes would increase progressively, indicating miR-21-5p could be a potential biomarker of T1D (107). Fortunately, miRNAs in exosomes of some AIDs show high specificity and sensitivity for the diagnosis of AIDs disease process (90,96,102,109). However, the sample sizes in these studies were small (N<70), and we should further validate its sensitivity and specificity in a large population before using it as a biomarker of disease activity in AIDs.…”

“…However, the sample sizes in these studies were small (N<70), and we should further validate its sensitivity and specificity in a large population before using it as a biomarker of disease activity in AIDs. In addition, in the exosomes of AIDs, the pathogenic mechanism of some miRNAs with high diagnostic value of disease process has not been studied (90,96,109). There is no doubt that these pathogenic exosomes provide targets for the treatment of AIDs, but how to identify, isolate and remove these pathogenic "bad" exosomes will become another problem.…”