Serum MicroRNA Signature as a Diagnostic and Therapeutic Marker in Patients with Psoriatic Arthritis

Abstract

Objective MicroRNAs (miRNAs) are small endogenous regulatory RNA molecules, which have emerged as potential therapeutic targets and biomarkers in autoimmunity. Here, we investigated serum miRNA levels in Psoriatic arthritis (PsA) patients and further assessed a serum miRNA signature in therapeutic responder versus non-responder PsA patients. Methods Serum samples were collected from healthy controls (n=20) and PsA (n=31) and clinical demographics obtained. To examine circulatory miRNA in serum from HC and PsA patients a focussed immunology miRNA panel was analysed utilising a miRNA Fireplex assay. MiRNA expression was further assessed in responders versus non responders according to the EULAR response criteria Results Six miRNA (miR-221-3p, miR-130a-3p, miR-146a-5p, miR-151-5p, miR-26a-5p and miR-21-5p) were significantly higher in PsA compared to healthy controls (all p<0.05), with high specificity and sensitivity determined by receiver operating characteristic (ROC) curve analysis. Analysis of responder vs non-responders demonstrated higher baseline levels of miR- 221-3p, miR-130a-3p, miR-146a-5p, miR-151-5p and miR-26a-5p were associated with therapeutic response. Conclusion This study identified a six-serum microRNA signature that could be attractive candidates as non-invasive markers for PsA, and may help to elucidate the disease pathogenesis.

Serum miRNA Signature in Rheumatoid Arthritis and “At-Risk Individuals”

“…In order to compare the miRNA signature of RA and healthy sera, a specific panel of 68 miRNAs, known to be associated with immune dysfunction ( 14 ), was selected for multiplex miRNA analysis of serum from HC donors (N = 20) and RA patients that were naïve to treatment (N = 50). The FirePlex Analysis Workbench software was used to analyze and determine significant differences in miRNA expression, between RA and HC, and generate heatmaps.…”

“…Patient blood was collected in anticoagulant-free tubes and processed within 1 h of collection as follows: samples were centrifuged at 2,000 g for 10 min at room temperature to isolate the serum which was transferred to RNase- and DNase-free tubes. The FirePlex Circulating miRNA Immunology Fixed Panel ( Supplementary Table 3 ) (Abcam, Cambridge, UK) was selected for multiplex miRNA analysis of 68 miRNAs in serum as previously described ( 14 ). This panel was utilized to focus on miRNA associated with immune cell dysfunction in autoimmune diseases, and included miRNA that have been previously identified to be associated with RA, either by expression in serum/tissue/cells or associated with immune cell/synovial fibroblast dysfunction ( 14 , 17 – 20 , 24 ).…”

“…The FirePlex Circulating miRNA Immunology Fixed Panel ( Supplementary Table 3 ) (Abcam, Cambridge, UK) was selected for multiplex miRNA analysis of 68 miRNAs in serum as previously described ( 14 ). This panel was utilized to focus on miRNA associated with immune cell dysfunction in autoimmune diseases, and included miRNA that have been previously identified to be associated with RA, either by expression in serum/tissue/cells or associated with immune cell/synovial fibroblast dysfunction ( 14 , 17 – 20 , 24 ). Specifically we utilized this technique to assess the utility of the FirePlex as a high-throughput platform which can reliably quantify miRNA directly from crude biofluids with minimal sample preparation, thus reducing the RNA isolation-induced variability.…”

“…In addition to their localization within the cell, miRNAs are also present in extracellular fluids such as serum, plasma, and synovial fluid, and can be transported to target cells and tissues through the circulation, thus acting as signaling molecules for intercellular communication ( 10 – 12 ). These are more stable than cellular miRNAs and many have been reported as potential biomarkers for disease ( 13 , 14 ). Early diagnosis is crucial in halting the development of RA in order to prevent disability and maintain quality of life, and systemic inflammation is often detected in individuals prior to the clinical presentation of arthritis.…”

Measurements Methods for the Development of MicroRNA-Based Tests for Cancer Diagnosis

“…An advantage of this method is that it allows quantification directly from biofluids, starting from 10 μL, without the need for miRNA extraction or processing. The Fireplex platform has been recently applied for miRNA biomarker studies in different diseases, including psoriatic arthritis, Duchenne muscular dystrophy and cardiac disease, although an extensive literature is still lacking due to the novelty of the technique [ 150 , 151 , 152 ]. In the oncologic field, Leng et al showed a good concordance of measures obtained with Fireplex, performed directly on plasma samples, compared to the measures obtained by qPCR, upon quantification of 11 specific miRNAs as putative lung cancer biomarkers [ 153 ].…”