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DNA methylation regulates the expression of CXCL12 in rheumatoid arthritis synovial fibroblasts

Genes & Immunity volume 12, pages 643–652 (2011)Cite this article

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Abstract

In the search for specific genes regulated by DNA methylation in rheumatoid arthritis (RA), we investigated the expression of CXCL12 in synovial fibroblasts (SFs) and the methylation status of its promoter and determined its contribution to the expression of matrix metalloproteinases (MMPs). DNA was isolated from SFs and methylation was analyzed by bisulfite sequencing and McrBC assay. CXCL12 protein was quantified by enzyme-linked immunosorbent assay before and after treatment with 5-azacytidine. RASFs were transfected with CXCR7-siRNA and stimulated with CXCL12. Expression of MMPs was analyzed by real-time PCR. Basal expression of CXCL12 was higher in RASFs than osteoarthritis (OA) SFs. 5-azacytidine demethylation increased the expression of CXCL12 and reduced the methylation of CpG nucleotides. A lower percentage of CpG methylation was found in the CXCL12 promoter of RASFs compared with OASFs. Overall, we observed a significant correlation in the mRNA expression and the CXCL12 promoter DNA methylation. Stimulation of RASFs with CXCL12 increased the expression of MMPs. CXCR7 but not CXCR4 was expressed and functional in SFs. We show here that RASFs produce more CXCL12 than OASFs due to promoter methylation changes and that stimulation with CXCL12 activates MMPs via CXCR7 in SFs. Thereby we describe an endogenously activated pathway in RASFs, which promotes joint destruction.

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Acknowledgements

We thank Mrs Maria Comazzi and Mr Ferenc Pataky for their excellent technical work. We also thank our collaborators Professor C Buckley and Dr A Filer for providing us with the human lung fibroblast cultures. This work was supported by the Articulum Fellowship Program (YR), the Institute of Arthritis Research, Epilanges, Switzerland, and by the European Community's FP6 (Autocure) and FP7 funding (Masterswitch). This publication reflects only the author's views. The European Community is not liable for any use that may be made of the information herein.

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Authors and Affiliations

  1. Center of Experimental Rheumatology and Zurich Center of Integrative Human Physiology (ZIHP), University Hospital Zurich, Zurich, Switzerland

    E Karouzakis, Y Rengel, A Jüngel, R E Gay, B A Michel, S Gay, M Neidhart & C Ospelt

  2. Schulthess Clinic, Zurich, Switzerland

    C Kolling

  3. Department of Clinical Immunology and Rheumatology, Academic Medical Center, University of Amsterdam, Amsterdam, The Netherlands

    P P Tak & C Ospelt

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  1. E Karouzakis
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Correspondence to E Karouzakis.

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Karouzakis, E., Rengel, Y., Jüngel, A. et al. DNA methylation regulates the expression of CXCL12 in rheumatoid arthritis synovial fibroblasts. Genes Immun 12, 643–652 (2011). https://doi.org/10.1038/gene.2011.45

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