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Volume 3 Supplement 2

21st European Workshop for Rheumatology Research

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Patterns of differentially expressed genes in synovial tissue from RA and OA patients and from normal joints


To identify key genes in the pathomechanism of rheumatoid arthritis (RA), synovial tissues from RA, osteoarthritis (OA) and from normal joints (ND) were compared by a subtractive hybridization technique, the representational difference analysis (RDA).


Synovial tissues from 3 RA, 3 OA patients and 5 normal joints were selected according to their disease-characteristic immunhistochemical findings and to their expression of high versus low levels of inflammatory (IL-1β, TNF-α) and destructive markers (MMP-1, MMP-3) as determined by semiquantitative RT-PCR. Pooled mRNA from RA, OA and normal tissues was transcribed, digested by a 4-base-cutter, ligated to adapter-primers and amplified to form representational amplicons. Subtractive hybridizations were performed by different protocols: 1. the OA amplicon (driver) was subtracted from the RA representation (tester); 2. the RA (driver) from the ND (tester) and 3. the ND (driver) from the OA rep-resentaion (tester). Using primers specific for the corresponding tester, the difference-products were selectively amplified, cloned, sequenced and compared to published sequences in the Genebank. Differential expression of identified genes was validated by semiquantitative RT-PCR.


Approximately 150 genes were found to be differentially expressed in RA synovial tissue as compared to OA or ND tissues respectively, or in OA tissues as compared to ND. Interestingly, some genes were identified to be overexpressed in both groups: RA (i.e. difference-product from RA minus OA) and OA (OA minus ND), indicating rather an association to general joint destruction than to RA-specific mechanism. Other genes were found to be differentially expressed only in the RA representation. 30 of the differentially expressed genes identified from each disease group were analyzed in synovial tissues from further 20 RA, 20 OA patient and 20 normal joints. The expression of some genes showed either a significant correlation to those of inflammatory genes (IL-1β and TNF-α) or to those of destructive markers (MMP-3).


The analysis of differential gene expression in chronic joint diseases is a promising approach to identify deregulation of the inflammatory network to explain the inappropriate immune response with autoaggessive outcome. Furthermore a pattern of genes is generated which is specifically or preferentially expressed in RA. Such patterns will be of diagnostic value, especially for disease characterization, longitudinal studies and analysis of therapeutic effects.

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Ungethüm, U., Häupl, T., Zacher, J. et al. Patterns of differentially expressed genes in synovial tissue from RA and OA patients and from normal joints. Arthritis Res Ther 3 (Suppl 2), P045 (2001).

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