Session Title: Rheumatoid Arthritis - Animal Models
Session Type: Abstract Submissions (ACR)
Epigenetic regulation plays an important role in inflammatory arthritis, including rheumatoid arthritis (RA). Histone deacetylase inhibitor (HDACi) has been recently reported to have therapeutic effect in collagen induced arthritis (CIA). CKD-L is a new HDAC6i developed by Chong Kun Dang Pharmaceutical Corporation. We investigated the therapeutic effect of selective HDAC6 inhibitors (CKD-L and Tubastatin A) on CIA, peripheral blood mononuclear cells (PBMCs) and regulatory T (Treg) cells.
CIA was induced by bovine type II collagen (CII) in DBA/1 mouse. Mice were treated with vehicle (n=10), CKD-L (15, 30 mg/kg, n=10, respectively) or Tubastatin A (30 mg/kg, n=10) by subcutaneous injection every day for 18 days. Arthritis score was assessed twice weekly after the onset of arthritis. Histological analysis was performed by H&E stain. CD4+CD25– cells were isolated from C57BL/6 mice spleen and activated with anti-CD3/CD28 beads, TGF-β and HDAC6i (1~10 μM) for 6 days. Cytotoxic T lymphocyte associated protein 4 (CTLA-4) expression in induced Treg (iTreg) cells was analysed by flow cytometry. Natural Treg (nTreg) cells and CD4+CD25– (Teff) cells were isolated from C57BL/6 mice spleen. nTreg cells were incubated with anti-CD3/CD28 bead, HDAC6i (200~2000 nM) and carboxyfluorescein succinimidyl ester (CFSE)-labeled Teff cells for 3 days (Treg:Teff ratio=1:2). Proliferation of Teff cells was analysed by flow cytometry. RA PBMCs were stimulated with lipopolysaccharide 100 ng/ml and HDAC6i (10-5000 nM) for 24 hours. Multiplex cytokine assay was performed with supernatant. RA CD4+CD25– cells were cultured with anti-CD3 Ab, anti-CD28 Ab, IL-2, TGF-β and 1,25(OH)2VD3 for 5 days. iTreg cells were incubated for 3 days with CFSE-stained Teff cells in the presence of anti-CD3/CD28 beads and HDAC6i (10-5000 nM). Proliferation of Teff cells was analysed by flow cytometry.
In CIA, CKD-L and Tubastatin A significantly reduced arthritis score and histological score. CTLA-4 expression in mouse iTreg cells was increased after treatment of CKD-L (P<0.001) and Tubastatin A (P<0.05). And mouse nTreg cells inhibited the proliferation of Teff cells after treatment with CKD-L (67.8 %) and Tubastatin A (68.3 %) compared to no treatment (80.1 %). In RA PBMC, TNF-α was decreased after treatment with CKD-L (P<0.001) and Tubastatin A (P<0.001). IL-10 was increased after treatment CKD-L (P=0.051) and Tubastatin A (P=0.083). In co-culture with iTreg cells and Teff cells, CKD-L efficiently inhibited the proliferation of Teff cells (13.9 %) compared to no treatment (32.6 %). Tubastatin A had no effect on proliferation of Teff cells (30.1%).
CKD-L was effective on the suppression of arthritis in CIA. CKD-L increased CTLA-4 expression and function of Treg cells. These results suggest that CKD have beneficial effect in the treatment of RA.
B. R. Oh,
Y. I. Choi,
H. J. Yoo,
J. K. Park,
E. Y. Lee,
E. B. Lee,
Y. W. Song,
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ACR Meeting Abstracts - https://acrabstracts.org/abstract/therapeutic-effect-of-a-novel-histone-deacetylase-6-inhibitor-ckd-l-on-collagen-induced-arthritis-and-peripheral-blood-mononuclear-cells-from-patients-with-rheumatoid-arthritis/