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Plasma and synovial fluid microRNAs as potential biomarkers of rheumatoid arthritis and osteoarthritis.

Author(s): Murata K, Yoshitomi H, Tanida S, Ishikawa M, Nishitani K, Ito H, Nakamura T

Publication: Arthritis Res Ther, 2010, Vol. 12, Page R86

PubMed ID: 20470394 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   The purpose of this study was to determine the stability of miRNA in plasma and synovial fluid following frozen storage at -20°C and freeze-thaw cycling. Plasma and synovial fluid from 30 patients with RA were stored at -20°C. RNA was extracted with Phenol-chloroform and miRNA was purified using the High Pure miRNA Isolation Kit.

   Summary of Findings:

   Storage of plasma or synovial fluid, at -20°C for up to 7 days, did not affect the quantified levels of miR-16 or miR-223. In contrast, levels of miR-16 and miR-223 in plasma and miR-223 in synovial fluid were significantly lower after 3 freeze-thaw cycles, than after 1 cycle (p<0.05, p<0.05 and p<0.001, respectively). Levels of miR-16 in plasma were also significantly lower after 8 freeze-thaw cycles than 1 freeze-thaw cycle (p<0.05).

   Biospecimens

   • Bodily Fluid - Synovial Fluid
   • Bodily Fluid - Plasma

   Preservative Types

   • Frozen

   Diagnoses:

   • Rheumatoid Arthritis

   Platform:

   Analyte	Technology Platform
   RNA	Real-time qRT-PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Storage	Storage duration	1 day 3 days 7 days
   Real-time qRT-PCR Specific	Targeted nucleic acid	miR-16 miR-223
   Storage	Freeze/thaw cycling	1 cycle 3 cycles 8 cycles

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2. Study Purpose

   The purpose of this study was to compare miRNA expression between synovial fluid and plasma and to identify potential miRNA biomarkers of RA and OA. K2EDTA plasma and synovial fluid were obtained from 20 patients with RA and 22 patients with OA and was stored frozen at -20°C.  Additionally during surgery, synovial tissues (3 RA and 3 OA), FLS (4 RA and 5 OA) and peripheral blood (3 RA, 3 OA and 3 from healthy controls) were obtained and cultured. RNA was extracted with Phenol-chloroform and miRNA was purified using the High Pure miRNA Isolation Kit.

   Summary of Findings:

   Mean concentrations of miR-16, miR-132, miR-146a, and miR-223 were significantly lower in synovial fluid than plasma both in patients with RA (p<0.01, p<0.05, p<0.01 and p<0.05, respectively) and in patients with OA (p<0.01, p<0.01, p<0.01 and p<0.01, respectively). While generally miRNA levels in between plasma and synovial fluid were not correlated , in OA patients levels of miR-223in plasma and synovial fluid were modestly correlated (r=0.50, p=0.01). Culture of FLS, PBMC, synovial tissues and synovial fluid MNCs revealed different secretion patterns. FLS and synovial tissue secreted large quantities of miR-132 and smaller quantities of miR-223, while MNCs secreted moderate quantities of miR-223 and miR-155 and smaller quantities of miR-16, miR-132 and miR146a. Based on these findings the authors conclude that synovial tissues are the main source of synovial fluid miRNA, but that expression of miR-223 is influenced by MNCs and thus reflects the condition of the joint. Plasma levels of miR-132 were higher in healthy controls than in patients with OA or RA (p<0.01, both) and plasma levels of miR-16 were higher in healthy controls than in patients with OA (p<0.05). Although plasma miRNA levels were comparable between OA and RA patients, synovial fluid from patients with RA had higher levels of miR-16, miR-146a miR-155 and miR-223 than synovial fluid from patients with OA (p<0.01, p<0.05, p<0.05 and p<0.05, respectively). Synovial fluid miRNA levels were not correlated with any clinical variables of RA. However, plasma miR-16, miR-146a, miR-155, and miR-223 levels were modestly and inversely correlated with TJC (r=-0.55, p<0.01; r=-0.54, p<0.01; r=-0.45, p=0.03; and r=-0.49, p=0.02; respectively).  The synovial fluid/peripheral blood ratio of miR-16, miR-132 and miR-146a were modestly to strongly inversely correlated with TJC (r=0.71, p<0.01; r=0.67, p<0.01; and r=0.80, p<0.01, respectively).

   Biospecimens

   • Bodily Fluid - Plasma
   • Bodily Fluid - Synovial Fluid
   • Cell - White Cells
   • Tissue - Other
   • Bodily Fluid - Blood

   Preservative Types

   • Frozen

   Diagnoses:

   • Normal
   • Rheumatoid Arthritis
   • Osteoarthritis

   Platform:

   Analyte	Technology Platform
   RNA	Real-time qRT-PCR

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Preaquisition	Diagnosis/ patient condition	Healthy Rheumatoid arthritis Osteoarthritis
   Real-time qRT-PCR Specific	Targeted nucleic acid	miR-16 miR-132 miR-146a miR-155 miR-223
   Biospecimen Acquisition	Biospecimen location	Synovial fluid Plasma Synovial tissue Synovial mononuclear cells PBMNC

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