Location of Repository

DNA demethylation at specific CpG sites in the IL1B promoter in response to inflammatory cytokines in human articular chondrocytes

By Ko Hashimoto, Richard O.C. Oreffo, Marc B. Gibson, Mary B. Goldring and Helmstrud I. Roach

Abstract

Objective: to determine whether changes in the DNA methylation status in the promoter region of the gene encoding interleukin-1? (IL-1?) account for expression of IL1B mRNA after long-term treatment of human articular chondrocytes with inflammatory cytokines.<br/><br/>Methods: IL-1? or TNF? combined with oncostatin M (OSM), or 5-aza-deoxycytidine (5-aza- dC) were added twice weekly for 4-5 weeks to primary cultures of normal human articular chondrocytes, obtained from patients with a femoral neck fracture. Expression of MMP13, IL1B??TNFA and DNMT1 were determined by SybrGreen-based qRT-PCR on genomic DNA and total RNA extracted from the same sample before and after culture. Bisulfite modification was used to identify which CpG sites in the IL1B promoter showed differential methylation between expressing and non-expressing cells. The percentages of cells that were methylated at that critical CpG site (-299bp) were quantified by a method that depended on methylation-sensitive restriction enzymes and real-time PCR. Secretion of IL-1? was assessed by enzyme-linked immunoabsorbent assay (ELISA) of the culture media.<br/><br/>Results: healthy chondrocytes did not express IL1B mRNA, but the levels were increased 5-fold by 5-aza-dC and 100- to 1000-fold by or TNF?/OSM. The % CpG methylation was decreased by 5-aza-dC, but reduced considerably more by IL-1? and almost abolished by TNF?/OSM. The mRNA was translated into protein in cytokine-treated chondrocytes.<br/><br/>Conclusion: these novel findings indicate that inflammatory cytokines can change DNA methylation status at key CpG sites, resulting in long-term induction of IL1B in human articular chondrocytes.<br/

Topics: Q1, R1, RZ
Year: 2009
OAI identifier: oai:eprints.soton.ac.uk:72820
Provided by: e-Prints Soton

Suggested articles

Preview

Citations

  1. (1996). A role for nuclear NF-B in B-cell-specific demethylation of the Ig locus. doi
  2. Activation of interleukin-1 signaling cascades in normal and osteoarthritic articular cartilage. doi
  3. (2003). Adenoviral gene transfer of interleukin-1 in combination with oncostatin M induces significant joint damage in a murine model. doi
  4. (1990). An inducible enhancer controls the expression of the human interleukin 1 gene. Cell Growth Differ
  5. (2005). Association between the abnormal expression of matrix-degrading enzymes by human osteoarthritic chondrocytes and demethylation of specific CpG sites in the promoter regions. Arthritis Rheum doi
  6. Autocrine induction of the human pro-IL-1 gene promoter by IL-1 in monocytes. doi
  7. Cellular and epigenetic features of a young healthy and a young osteoarthritic cartilage compared with aged control and OA cartilage. doi
  8. (2006). Comparative analysis of gene expression profiles in intact and damaged regions of human osteoarthritic cartilage. Arthritis Rheum doi
  9. (1997). DBA/1 mice expressing the human TNF-a transgene develop a severe, erosive arthritis: characterization of the cytokine cascade and cellular composition.
  10. Defining the roles of inflammatory and anabolic cytokines in cartilage metabolism. doi
  11. (2002). DNA methylation and the regulation of gene transcription. Cell Mol Life Sci doi
  12. DNA methylation patterns and epigenetic memory. doi
  13. Dynamic protein associations define two phases of IL-1 transcriptional activation. doi
  14. Epigenetic regulation of gene expression: how the genome integrates intrinsic and environmental signals. doi
  15. (2007). Epigenetic regulation of leptin affects MMP-13 expression in osteoarthritic chondrocytes: possible molecular target for osteoarthritis therapeutic intervention. Ann Rheum Dis doi
  16. (2009). Expression of ADAMTS-4 by chondrocytes in the surface zone of human osteoarthritic cartilage is regulated by epigenetic DNA demethylation. Rheumatol Int doi
  17. (2004). Goldring MB. The role of cytokines in cartilage matrix degeneration in osteoarthritis. Clin Orthop Relat Res doi
  18. IL-1, but not BMP-7, leads to a dramatic change in the gene expression pattern of human adult articular chondrocytes— portraying the gene expression pattern in two donors. doi
  19. (1999). Immunohistological analysis of cytokine expression in human osteoarthritic and healthy cartilage.
  20. Improved quantification of DNA methylation using methylation-sensitive restriction enzymes and real-time PCR. Epigenetics 2007;2:86–91. doi
  21. (2007). In vivo osteoarthritis target validation utilizing genetically-modified mice. Curr Drug Targets doi
  22. (2004). Increased AP-1 and NF-B activation and recruitment with the combination of the proinflammatory cytokines IL-1, tumor necrosis factor and IL-17 in rheumatoid synoviocytes. Arthritis Res Ther
  23. Induction of matrix metalloproteinase-13 gene expression by TNF- is mediated by MAP kinases, AP-1, and NF-B transcription factors in articular chondrocytes. doi
  24. (2006). Interleukin-1 in combination with oncostatin M upregulates multiple genes in chondrocytes: implications for cartilage destruction and repair. Arthritis Rheum doi
  25. Intracellular interleukin-1 functionally interacts with histone acetyltransferase complexes. doi
  26. (2006). Large-scale gene expression profiling reveals major pathogenetic pathways of cartilage degeneration in osteoarthritis. Arthritis Rheum doi
  27. (2001). Matrix metalloproteinase and proinflammatory cytokine production by chondrocytes of human osteoarthritic cartilage: associations with degenerative changes. Arthritis Rheum doi
  28. Mechanisms of epigenetic inheritance. doi
  29. Methyl-CpG binding domain 1 (MBD1) interacts with the Suv39h1-HP1 heterochromatic complex for DNA methylation-based transcriptional repression. doi
  30. (2009). Methylation of the OP-1 promoter: potential role in the age-related decline in OP-1 expression in cartilage. Osteoarthritis Cartilage doi
  31. (1995). Monocyte expression of the human prointerleukin 1 gene (IL1B) is dependent on promoter sequences which bind the hematopoietic transcription factor Spi-1/PU.1. Mol Cell Biol
  32. (1999). Multiple signals converging on NF-B. doi
  33. Nuclear factor-B is a critical mediator of Ste20-like proline-/alanine-rich kinase regulation in intestinal inflammation. doi
  34. (2003). Oncostatin M in combination with tumor necrosis factor induces cartilage damage and matrix metalloproteinase expression in vitro and in vivo. Arthritis Rheum doi
  35. (2006). Osteoarthritis: pathobiology-targets and ways for therapeutic intervention. Adv Drug Deliv Rev doi
  36. Persistent interleukin-1 signaling causes long term activation of NFB in a promoter-specific manner in human glial cells. doi
  37. Phenotypic plasticity and the epigenetics of human disease. doi
  38. Signaling pathways implicated in oncostatin M-induced aggrecanase-1 and matrix metalloproteinase-13 expression in human articular chondrocytes. doi
  39. Stability and flexibility of epigenetic gene regulation in mammalian development. doi
  40. (2008). The additive effect of individual genes in predicting risk of knee osteoarthritis. Ann Rheum Dis doi
  41. The complex language of chromatin regulation during transcription. doi
  42. (2006). The effect of IL-1 on the expression of inflammatory cytokines and their receptors in human chondrocytes. Life Sci doi
  43. (1995). The effects of 5-azacytidine and 5-azadeoxycytidine on chromosome structure and function: implications for methylationassociated cellular processes. Pharmacol Ther doi
  44. (1993). The human prointerleukin 1 gene requires DNA sequences both proximal and distal to the transcription start site for tissue-specific induction.
  45. The methyl-CpG-binding protein MeCP2 links DNA methylation to histone methylation. doi
  46. (2007). The pathogenesis of osteoarthritis. In: Bronner F and Farach-Carson MC editors. Bone and osteoarthritis. Topics in bone biology. doi
  47. (2004). The regulation of chondrocyte function by proinflammatory mediators: prostaglandins and nitric oxide. Clin Orthop Relat Res doi

To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.