Does epigenetics play a role in the pathology of osteoarthritis?
Yamada, N., Cheung, K.S.C., Tilley, S., Clarke, N.M.P., Oreffo, R.O.C., Kokobun, S., Bronner, F. and Roach, H.I. (2006) Does epigenetics play a role in the pathology of osteoarthritis? Journal of Bone and Joint Surgery, 88-B, (Supplement III), p.403.
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Idiopathic osteoarthritis (OA) is a complex, late-onset disease whose causes are still unknown. In spite of tremendous efforts, the search for the genes pre-disposing towards osteoarthritis has so far met with little success. We hypothesize that epigenetic changes play a major role in the pathology of OA. Epigenetics refers to stable, heritable, but potentially reversible modifications of gene expression that do not involve mutations in the DNA sequence, for example DNA methylation or histone modification. Epigenetic changes are gene and cell-type specific, may arise sporadically with increasing age or be provoked by environmental factors. To investigate whether epigenetic changes are significant factors in OA, we examined the DNA methylation status of the promoter regions of three genes that are expressed by OA, but not by normal, articular chondrocytes, namely MMP-3 (stromelysin-1), MMP-9 (gelatinase B) and MMP-13 (collagenase3). We hypothesized that these genes are silenced in normal chondrocytes by methylation of the cytosines of CpG dinucleotides in the respective promoter regions, but that abnormal expression is associated with a de-methylation, leading to eunsilencing f of gene expression. Cartilage was obtained from the femoral heads of 16 OA and 10 femoral neck fracture (#NOF) patients, which served as controls due to the inverse relationship between osteoporosis and OA. The cartilage was milled in a freezer mill with liquid nitrogen, DNA was extracted with a Qiagen kit, digested with methylation sensitive restriction enzymes, followed by PCR amplification. These enzymes will cut at their specific cleavage sites only if the CpGs is not methylated and thus allow us to determine methylation status of specific CpG sites.
Results. Less than 5% of the chondrocytes in superficial layer from #NOF cartilage expressed degradative enzymes, whereas all cloned chondrocytes from advanced-stage OA cartilage were immunopositive. The overall % of CpG demethylation in the promoters of control patients (whose chondrocytes did not express the enzymes) was 20.1%, whereas 48.6% of CpG sites were demethylated in degradative chondrocytes of OA patients (p<0.001). For MMP-13, the increase in demethylation between control and OA was from 4 ..20%; for MMP-9 from 47 ..81% and for MMP-3 from 30 ..57%. However, not all available CpG sites were equally demethylated. Some sites were uniformly methylated in both OA and controls, others were demethylated even in controls. However, there was at least one crucial site for each degradative enzyme, where the differences in the degree of methylation were greatest and statistically different. These sites were at –110 for MMP-13; –36 for MMP-9; –635 for MMP-3. There was no relation between the % demethylation and the patient fs age and no apparent difference between males and females.
Conclusions: We have demonstrated an association between abnormal gene expression of MMP-3, MMP-9 and MMP-13 and promoter DNA demethylation. This epigenetic dysregulation of genes appeared to be clonally inherited by daughter cells and may be typical for osteoarthritis and other complex, late-onset diseases.
Correspondence should be addressed to Mr Carlos Wigderowitz, Honorary Secretary BORS, University Dept of Orthopaedic & Trauma Surgery, Ninewells Hospital & Medical School, Dundee DD1 9SY.
None of the authors have received anything of value from a commercial or other party related directly or indirectly to the subject of the presentation
|Subjects:||R Medicine > RB Pathology
R Medicine > RF Otorhinolaryngology
Q Science > QH Natural history > QH426 Genetics
|Divisions:||University Structure - Pre August 2011 > School of Medicine
|Date Deposited:||14 Nov 2008|
|Last Modified:||02 Mar 2012 11:32|
|Contributors:||Yamada, N. (Author)
Cheung, K.S.C. (Author)
Tilley, S. (Author)
Clarke, N.M.P. (Author)
Oreffo, R.O.C. (Author)
Kokobun, S. (Author)
Bronner, F. (Author)
Roach, H.I. (Author)
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