J Rheum Dis.  2020 Jan;27(1):61-67. 10.4078/jrd.2020.27.1.61.

MicroRNA-10b Plays a Role in Bone Formation by Suppressing Interleukin-22 in Ankylosing Spondylitis

  • 1Department of Rheumatology, Research Institute of Medical Sciences, Chonnam National University Medical School and Hospital, Gwangju, Korea. ktj1562@chonnam.ac.kr
  • 2Department of Rheumatology, Hanyang University Hospital for Rheumatic Diseases, Seoul, Korea.


The microRNA (miR)-10b is the T helper (Th) 17 cell specific in patients with ankylosing spondylitis (AS). The interleukin (IL)-22, which is closely related to Th17 cells, has been implicated in the regulation of new bone formation in experimental models. Therefore, the aim of this study was to evaluate whether miR-10b affects bone formation via the IL-22 pathway in AS.
Primary CD4+ T cells from AS were purified and transfected with miR-10b, anti-miR-10b, or scramble. Cell-surface markers and cytokine expression were analyzed by flow cytometry and enzyme-linked immunosorbent assay. Primary bone-derived cells (BdCs) from the facet joints of the spine were isolated, then osteogenic differentiation of primary BdCs was performed. We assessed alkaline phosphatase (ALP) activity and staining of BdCs at early time points. Alizarin red S staining of BdCs was performed at late time points.
Overexpression of miR-10b reduced both IL-22 producing cell frequencies and cytokine production in T cells from the patients with AS. The IL-22 significantly increased ALP staining and bone mineralization. The ALP promotor activity of AS-BdCs was notably higher for the IL-22 concentration. The supernatants of the miR-10b overexpression group suppressed ALP activity on osteogenic progenitor cells from the facet joints of the spine in patients with AS.
Our data suggest that miR-10b suppresses IL-22 production, which was involved in osteogenic proliferation in AS. Therefore, miR-10b might be a potential therapeutic candidate for regulation of new bone formation in patients with AS.


Ankylosing spondylitis; MicroRNA; Th17 cell; Osteogenesis

MeSH Terms

Alkaline Phosphatase
Calcification, Physiologic
Enzyme-Linked Immunosorbent Assay
Flow Cytometry
Models, Theoretical
Spondylitis, Ankylosing*
Stem Cells
Th17 Cells
Zygapophyseal Joint
Alkaline Phosphatase


  • Figure 1 MicroRNA (miR)-10b suppresses interleukin (IL)-22 cytokine production. Frequency of IL-22 producing cells and production of IL-22 in CD4 T cells from ankylosing spondylitis (AS) transfected with miR-10b mimic, anti-miR-10b, or miR-scramble and stimulated with anti-CD2/3/28-coated beads for 72 hours. A representative flow cytometry result is shown for each group (A). Overexpression of miR-10b reduced IL-22 producing cell frequencies and cytokine production in T cells from the patients with AS. Whereas, inhibition of miR-10b significantly increased the expression of IL-22 cytokine (B, C). APC: allophycocyanin, PE-Cy7: phycoerythrin-Cy7.

  • Figure 2 Interleukin (IL)-22 promotes osteoblastogenesis. Primary bone-derived cells (BdCs) from ankylosing spondylitis (AS) were isolated and cultured. Alkaline phosphatase (ALP) activity and staining were assessed at early time points after stimulation with osteogenic medium (OM). Alizarin red S (ARS) staining was done at late time points after stimulation with OM (A). The ALP promotor and intercellular ALP activity of AS-BdCs depending on IL-22 concentration were measured (B, C).

  • Figure 3 MicroRNA (miR)-10 suppresses alkaline phosphatase (ALP) activity on osteoprogenitor cells in ankylosing spondylitis (AS). Supernatants from T cells, which had been transfected with miR-10b mimic or scramble, were added on the primary bone-derived cells bone-derived cells from the facet joint in AS.

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