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Overexpression of microRNA-223 in rheumatoid arthritis synovium controls osteoclast differentiation

  • Original Article
  • Published:
Modern Rheumatology

Abstract

Objectives

MicroRNAs, a class of noncoding RNAs, play roles in human diseases. MicroRNA-223 (miR-223) is reported to play critical roles in osteoclastogenesis. The purpose of this study was to analyze the expression pattern of miR-223 in rheumatoid arthritis (RA) synovium and examine the suppression of osteoclastogenesis from human peripheral blood mononuclear cells (PBMC) by overexpression of miR-223.

Methods

Expression of miR-223 in synovium from RA patients was analyzed by quantitative reverse transcription polymerase chain reaction (RT-PCR) and section in situ hybridization. MiR-223 was overexpressed in an osteoclastogenesis coculture system with PBMC and RA synovial fibroblast. At 3 weeks after transfection of double-stranded miR-223, the formation of tartrate-resistant acid phosphatase (TRAP)-stained multinucleated cells was analyzed to evaluate the inhibitory effect of miR-223 on osteoclastogenesis.

Results

MiR-223 was more highly expressed in RA synovium than in osteoarthritis (OA) synovium due to the increased number of miR-223-positive cells in RA synovium. MiR-223 was expressed in the superficial and sublining layers, and macrophages, monocytes, and CD4 T cells also expressed miR-223. The number of TRAP-positive multinucleated cells was significantly decreased by overexpression of miR-223 in a dose-dependent manner. The expression of osteoclastogenesis marker genes was significantly down-regulated by miR-223 overexpression.

Conclusion

MiR-223 is intensely expressed in RA synovium, and overexpression of miR-223 suppresses osteoclastogenesis in vitro. This study demonstrates the possibility of gene therapy with miR-223 to treat bone destruction in RA patients.

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Abbreviations

miR:

Micro ribonucleic acid

PBMC:

Peripheral blood mononuclear cells

RA:

Rheumatoid arthritis

RASF:

RA synovial fibroblasts

siRNA:

Small interfering RNA

TRAP:

Tartrate-resistant acid phosphatase

RANKL:

Receptor activator of nuclear factor-κB ligand

OPG:

Osteoprotegerin

PCR:

Polymerase chain reaction

TNF-α:

Tumor necrosis factor-α

IL-1:

Interleukin-1

IL-6:

Interleukin-6

RISC:

RNA-induced silencing complex

NFATc1:

Nuclear factor of activated T cell-1

NFI-A:

Nuclear factor I-A

C/EBP:

CCAAT/enhancer binding protein

M-CSF:

Macrophage colony-stimulating factor

DPBS-E:

Dulbecco’s phosphate buffered saline–EDTA

EDTA:

Ethylene diamine tetra acetic acid

UTR:

Untranslated region

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Acknowledgments

This work was supported in part by the Japan Rheumatism Foundation.

Conflict of interest

None.

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Authors and Affiliations

  1. Programs for Applied Biomedicine, Division of Clinical Medical Science, Department of Orthopedic Surgery, Graduate School of Biomedical Sciences, Hiroshima University, 1-2-3 Kasumi, Minami-ku, Hiroshima, Japan

    Hayatoshi Shibuya, Tomoyuki Nakasa, Nobuo Adachi, Yoshihiko Nagata, Masakazu Ishikawa, Masataka Deie, Osami Suzuki & Mitsuo Ochi

Authors
  1. Hayatoshi Shibuya
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  2. Tomoyuki Nakasa
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  3. Nobuo Adachi
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  4. Yoshihiko Nagata
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  7. Osami Suzuki
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  8. Mitsuo Ochi
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Correspondence to Hayatoshi Shibuya.

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Shibuya, H., Nakasa, T., Adachi, N. et al. Overexpression of microRNA-223 in rheumatoid arthritis synovium controls osteoclast differentiation. Mod Rheumatol 23, 674–685 (2013). https://doi.org/10.1007/s10165-012-0710-1

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  • DOI: https://doi.org/10.1007/s10165-012-0710-1

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