Background/Purpose:   The role of miRNAs in immune and inflammatory diseases is increasing exponentially. miRNAs are present in the human plasma in stable form. We proposed to test the hypothesis that patients with ankylosing spondylitis (AS) have an altered miRNA expression profile compared to controls.

Methods:   pared to controls. Methods: Sixtytwo patients ≥18 years of age with AS based on the modified New York classification criteria (grade 3 and 4 sacroiliitis) and 34 healthy (age, race and sex matched) controls were prospectively recruited. Subjects with active malignancy in last 5 years, rheumatoid arthritis, systemic lupus erythematosus and evidence of HIV or chronic hepatitis B or C infection were excluded. Patients and controls were screened, consented and peripheral blood samples (5 ml) were obtained. The samples were centrifuged at 400 g for seven minutes; plasma transferred to nuclease free tubes and stored at -20°C until analyses. ESR and CRP were measured using routine laboratory methods. Various validated Questionnaires to assess disease, functional activity and patient reported outcomes in AS were administered to the patients. Sixty-eight circulating miRNAs in the plasma of the study subjects were profiled by Firefly Multiplex Circulating miRNA Assay (Abcam, Cambridge, MA) using the Immunology Panel. The assay beads were scanned on BD Accuri C6 flow cytometer (BD Biosciences, San Jose, CA). Data were analyzed using Firefly Analyis Workbench (Abcam). Expression of ten selected miRNAs that were differentially expressed based on the multiplex assay was further validated by qPCR using TaqMan Advanced miRNA assay (ThermoFisher Scientific) following the manufacturer’s instructions. Briefly, Total RNA (including miRNA) was isolated from 100 μl of plasma from all the study subjects using the MagMAX mirVana Total RNA Isolation kit. cDNA was generated using 2 μl of purified total RNA with the TaqMan Advanced miRNA cDNA Synthesis kit. qPCR was then performed for each sample using 1 μL of diluted cDNA, TaqMan Advanced miRNA Assays, and Applied Biosystems TaqMan™ Fast Advanced Master Mix under fast cycling conditions. Reactions were run on StepOne Plus real-time PCR system. Data were analyzed on DataAssist v3.01 (ThermoFisher Scientific). Descriptive analyses included continuous variables (the mean ± SD) and the categorical variables(percentage).

Results:   Demographics, clinical characteristics and disease activity of the patients are in Table 1. Our data revealed that miR-30b-5p (p=0.005), miR-34a-5p (p=0.007), and miR-431-3p (p=0.014) were highly abundant in the plasma of AS patients compared to the levels prevalent in controls.

Table-1

Patient Demographics

Conclusion:   This study demonstrates that specific miRNAs are differentially expressed between AS patients and healthy subjects and thus can be developed as potential biomarkers.  This work was supported in part by USPHS/NIH grants (RO1 AT007373, RO1 AT005520, RO1 AR067056, R21 AR064890) and funds from North East Ohio Medical University to TMH.

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ACR Meeting Abstracts - https://acrabstracts.org/abstract/micrornas-mir-30b-5p-mir-34a-5p-and-mir-431-3p-are-highly-expressed-in-the-plasma-of-patients-with-ankylosing-spondylitis/