Detail information
ID ENOM00092
Accession GSE12860
Status Public on Sep 20, 2008
Title Antirheumatic Drug Response in Human Chondrocytes: Potential Molecular Targets to Stimulate Cartilage Regeneration
Organism Homo sapiens
Experiment type Expression profiling by array
Summary
Rheumatoid arthritis (RA) leads to progressive destruction of articular structures. Despite recent progress in controlling inflammation and pain, little cartilage repair has yet been observed. This in vitro study aims to determine the role of chondrocytes in RA-related cartilage destruction and antirheumatic drug-related regenerative processes. Human chondrocytes were three-dimensionally cultured in alginate beads. To determine the RA-induced gene expression pattern, human chondrocytes were stimulated with supernatant of RA synovial fibroblasts (RASF) and normal donor synovial fibroblasts (NDSF), respectively. To examine antirheumatic drug response signatures, human chondrocytes were stimulated with supernatant of RASF that have been treated with disease-modifying antirheumatic drugs (DMARD; azathioprine, sodium aurothiomalate, chloroquine phosphate, methotrexate), non-steroidal anti-inflammatory drugs (NSAID; piroxicam, diclofenac) or steroidal anti-inflammatory drugs (SAID; methylprednisolone, prednisolone). Genome-wide expression profiling with oligonucleotide microarrays was used to determine differentially expressed genes. Real-time RT-PCR and ELISA were performed for validation of microarray data. Following antirheumatic treatment, microarray analysis disclosed a reverted expression of 94 RA-induced chondrocyte genes involved in inflammation/NF-κB signalling, cytokine/chemokine activity, immune response, proliferation/differentiation and matrix remodelling. Hierarchical clustering analysis showed that treatment of RASF with the DMARD azathioprine, gold sodium thiomalate and methotrexate resulted in chondrocyte gene expression signatures that were closely related to the “healthy” pattern. Treatment with the SAID methylprednisolone and prednisolone strongly reverted the RA-related chondrocyte gene expression, in particular the expression of genes involved in inflammation/NF-κB and cytokine/chemokine activity. The NSAID piroxicam and diclofenac and the DMARD chloroquine phosphate had only moderate to marginal effects. Pathway analysis determined major mechanisms of drug action, for example pathways of cytokine-cytokine receptor interaction, TGF-β/TLR/Jak-STAT signalling and ECM-receptor interaction were targeted. This in vitro study provides a comprehensive molecular insight into the antirheumatic drug response signatures in human chondrocytes, thereby revealing potential molecular targets, pathways and mechanisms of drug action involved in chondrocyte regeneration. Thus, the present study may contribute to the development of novel therapeutic chondro-protective compounds and strategies.
Samples
GSM ID Sample info Characteristics Description
GSM322609 RASF stimulated chondrocytes rep 1 GSM253283 Human chondrocytes cultured in RPMI 1640 medium supplemented with 10% human serum, 100ng/ml amphotericin B, 100U/ml penicillin and 100µg/ml streptomycin for 2 passages, 3D cultured for 14 days in alginate beads in presence of 170µM L-ascorbic acid 2-phosphate and subsequently stimulated for 48h with supernatant of RASF cultured in the same medium Gene expression data of human chondrocytes stimulated with supernatant of RASF
GSM322610 RASF stimulated chondrocytes rep 2 GSM253284 Human chondrocytes cultured in RPMI 1640 medium supplemented with 10% human serum, 100ng/ml amphotericin B, 100U/ml penicillin and 100µg/ml streptomycin for 2 passages, 3D cultured for 14 days in alginate beads in presence of 170µM L-ascorbic acid 2-phosphate and subsequently stimulated for 48h with supernatant of RASF cultured in the same medium Gene expression data of human chondrocytes stimulated with supernatant of RASF
GSM322611 NDSF stimulated chondrocytes rep 1 GSM253285 Human chondrocytes cultured in RPMI 1640 medium supplemented with 10% human serum, 100ng/ml amphotericin B, 100U/ml penicillin and 100µg/ml streptomycin for 2 passages, 3D cultured for 14 days in alginate beads in presence of 170µM L-ascorbic acid 2-phosphate and subsequently stimulated for 48h with supernatant of NDSF cultured in the same medium Gene expression data of human chondrocytes stimulated with supernatant of NDSF
GSM322612 NDSF stimulated chondrocytes rep 2 GSM253286 Human chondrocytes cultured in RPMI 1640 medium supplemented with 10% human serum, 100ng/ml amphotericin B, 100U/ml penicillin and 100µg/ml streptomycin for 2 passages, 3D cultured for 14 days in alginate beads in presence of 170µM L-ascorbic acid 2-phosphate and subsequently stimulated for 48h with supernatant of NDSF cultured in the same medium Gene expression data of human chondrocytes stimulated with supernatant of NDSF
GSM322613 Chondrocytes stimulated with diclofenac treated RASF rep 1 Human chondrocytes cultured in RPMI 1640 medium supplemented with 10% human serum, 100ng/ml amphotericin B, 100U/ml penicillin and 100µg/ml streptomycin for 2 passages, 3D cultured for 14 days in alginate beads in presence of 170µM L-ascorbic acid 2-phosphate and subsequently stimulated for 48h with supernatant of RASF cultured in the same medium and treated with diclofenac for 48h Gene expression data of human chondrocytes stimulated with supernatant of RASF treated for 48h with diclofenac
GSM322614 Chondrocytes stimulated with diclofenac treated RASF rep 2 Human chondrocytes cultured in RPMI 1640 medium supplemented with 10% human serum, 100ng/ml amphotericin B, 100U/ml penicillin and 100µg/ml streptomycin for 2 passages, 3D cultured for 14 days in alginate beads in presence of 170µM L-ascorbic acid 2-phosphate and subsequently stimulated for 48h with supernatant of RASF cultured in the same medium and treated with diclofenac for 48h Gene expression data of human chondrocytes stimulated with supernatant of RASF treated for 48h with diclofenac
GSM322615 Chondrocytes stimulated with prioxicam treated RASF rep 1 Human chondrocytes cultured in RPMI 1640 medium supplemented with 10% human serum, 100ng/ml amphotericin B, 100U/ml penicillin and 100µg/ml streptomycin for 2 passages, 3D cultured for 14 days in alginate beads in presence of 170µM L-ascorbic acid 2-phosphate and subsequently stimulated for 48h with supernatant of RASF cultured in the same medium and treated with piroxicam for 48h Gene expression data of human chondrocytes stimulated with supernatant of RASF treated for 48h with piroxicam
GSM322616 Chondrocytes stimulated with prioxicam treated RASF rep 2 Human chondrocytes cultured in RPMI 1640 medium supplemented with 10% human serum, 100ng/ml amphotericin B, 100U/ml penicillin and 100µg/ml streptomycin for 2 passages, 3D cultured for 14 days in alginate beads in presence of 170µM L-ascorbic acid 2-phosphate and subsequently stimulated for 48h with supernatant of RASF cultured in the same medium and treated with piroxicam for 48h Gene expression data of human chondrocytes stimulated with supernatant of RASF treated for 48h with piroxicam
GSM322617 Chondrocytes stimulated with chloroquine phosphate (resochin) treated RASF rep 1 Human chondrocytes cultured in RPMI 1640 medium supplemented with 10% human serum, 100ng/ml amphotericin B, 100U/ml penicillin and 100µg/ml streptomycin for 2 passages, 3D cultured for 14 days in alginate beads in presence of 170µM L-ascorbic acid 2-phosphate and subsequently stimulated for 48h with supernatant of RASF cultured in the same medium and treated with chloroquine phosphate (resochin) for 48h Gene expression data of human chondrocytes stimulated with supernatant of RASF treated for 48h with chloroquine phosphate (resochin)
GSM322618 Chondrocytes stimulated with chloroquine phosphate (resochin) treated RASF rep 2 Human chondrocytes cultured in RPMI 1640 medium supplemented with 10% human serum, 100ng/ml amphotericin B, 100U/ml penicillin and 100µg/ml streptomycin for 2 passages, 3D cultured for 14 days in alginate beads in presence of 170µM L-ascorbic acid 2-phosphate and subsequently stimulated for 48h with supernatant of RASF cultured in the same medium and treated with chloroquine phosphate (resochin) for 48h Gene expression data of human chondrocytes stimulated with supernatant of RASF treated for 48h with chloroquine phosphate (resochin)
GSM322619 Chondrocytes stimulated with sodium aurothiomalate (tauredon) treated RASF rep 1 Human chondrocytes cultured in RPMI 1640 medium supplemented with 10% human serum, 100ng/ml amphotericin B, 100U/ml penicillin and 100µg/ml streptomycin for 2 passages, 3D cultured for 14 days in alginate beads in presence of 170µM L-ascorbic acid 2-phosphate and subsequently stimulated for 48h with supernatant of RASF cultured in the same medium and treated with sodium aurothiomalate (tauredon) for 48h Gene expression data of human chondrocytes stimulated with supernatant of RASF treated for 48h with sodium aurothiomalate (tauredon)
GSM322620 Chondrocytes stimulated with sodium aurothiomalate (tauredon) treated RASF rep 2 Human chondrocytes cultured in RPMI 1640 medium supplemented with 10% human serum, 100ng/ml amphotericin B, 100U/ml penicillin and 100µg/ml streptomycin for 2 passages, 3D cultured for 14 days in alginate beads in presence of 170µM L-ascorbic acid 2-phosphate and subsequently stimulated for 48h with supernatant of RASF cultured in the same medium and treated with sodium aurothiomalate (tauredon) for 48h Gene expression data of human chondrocytes stimulated with supernatant of RASF treated for 48h with sodium aurothiomalate (tauredon)
GSM322621 Chondrocytes stimulated with azathioprine (imurek) treated RASF rep 1 Human chondrocytes cultured in RPMI 1640 medium supplemented with 10% human serum, 100ng/ml amphotericin B, 100U/ml penicillin and 100µg/ml streptomycin for 2 passages, 3D cultured for 14 days in alginate beads in presence of 170µM L-ascorbic acid 2-phosphate and subsequently stimulated for 48h with supernatant of RASF cultured in the same medium and treated with azathioprine (imurek) for 48h Gene expression data of human chondrocytes stimulated with supernatant of RASF treated for 48h with azathioprine (imurek)
GSM322622 Chondrocytes stimulated with azathioprine (imurek) treated RASF rep 2 Human chondrocytes cultured in RPMI 1640 medium supplemented with 10% human serum, 100ng/ml amphotericin B, 100U/ml penicillin and 100µg/ml streptomycin for 2 passages, 3D cultured for 14 days in alginate beads in presence of 170µM L-ascorbic acid 2-phosphate and subsequently stimulated for 48h with supernatant of RASF cultured in the same medium and treated with azathioprine (imurek) for 48h Gene expression data of human chondrocytes stimulated with supernatant of RASF treated for 48h with azathioprine (imurek)
GSM322623 Chondrocytes stimulated with methotrexate treated RASF rep 1 Human chondrocytes cultured in RPMI 1640 medium supplemented with 10% human serum, 100ng/ml amphotericin B, 100U/ml penicillin and 100µg/ml streptomycin for 2 passages, 3D cultured for 14 days in alginate beads in presence of 170µM L-ascorbic acid 2-phosphate and subsequently stimulated for 48h with supernatant of RASF cultured in the same medium and treated with methotrexate for 48h Gene expression data of human chondrocytes stimulated with supernatant of RASF treated for 48h with methotrexate
GSM322624 Chondrocytes stimulated with methotrexate treated RASF rep 2 Human chondrocytes cultured in RPMI 1640 medium supplemented with 10% human serum, 100ng/ml amphotericin B, 100U/ml penicillin and 100µg/ml streptomycin for 2 passages, 3D cultured for 14 days in alginate beads in presence of 170µM L-ascorbic acid 2-phosphate and subsequently stimulated for 48h with supernatant of RASF cultured in the same medium and treated with methotrexate for 48h Gene expression data of human chondrocytes stimulated with supernatant of RASF treated for 48h with methotrexate
GSM322625 Chondrocytes stimulated with prednisolone (solu-decortin) treated RASF rep 1 Human chondrocytes cultured in RPMI 1640 medium supplemented with 10% human serum, 100ng/ml amphotericin B, 100U/ml penicillin and 100µg/ml streptomycin for 2 passages, 3D cultured for 14 days in alginate beads in presence of 170µM L-ascorbic acid 2-phosphate and subsequently stimulated for 48h with supernatant of RASF cultured in the same medium and treated with prednisolone (solu-decortin) for 48h Gene expression data of human chondrocytes stimulated with supernatant of RASF treated for 48h with prednisolone (solu-decortin)
GSM322626 Chondrocytes stimulated with prednisolone (solu-decortin) treated RASF rep 2 Human chondrocytes cultured in RPMI 1640 medium supplemented with 10% human serum, 100ng/ml amphotericin B, 100U/ml penicillin and 100µg/ml streptomycin for 2 passages, 3D cultured for 14 days in alginate beads in presence of 170µM L-ascorbic acid 2-phosphate and subsequently stimulated for 48h with supernatant of RASF cultured in the same medium and treated with prednisolone (solu-decortin) for 48h Gene expression data of human chondrocytes stimulated with supernatant of RASF treated for 48h with prednisolone (solu-decortin)
GSM322627 Chondrocytes stimulated with methylprednisolone (urbason) treated RASF rep 1 Human chondrocytes cultured in RPMI 1640 medium supplemented with 10% human serum, 100ng/ml amphotericin B, 100U/ml penicillin and 100µg/ml streptomycin for 2 passages, 3D cultured for 14 days in alginate beads in presence of 170µM L-ascorbic acid 2-phosphate and subsequently stimulated for 48h with supernatant of RASF cultured in the same medium and treated with methylprednisolone (urbason) for 48h Gene expression data of human chondrocytes stimulated with supernatant of RASF treated for 48h with methylprednisolone (urbason)
GSM322628 Chondrocytes stimulated with methylprednisolone (urbason) treated RASF rep 2 Human chondrocytes cultured in RPMI 1640 medium supplemented with 10% human serum, 100ng/ml amphotericin B, 100U/ml penicillin and 100µg/ml streptomycin for 2 passages, 3D cultured for 14 days in alginate beads in presence of 170µM L-ascorbic acid 2-phosphate and subsequently stimulated for 48h with supernatant of RASF cultured in the same medium and treated with methylprednisolone (urbason) for 48h Gene expression data of human chondrocytes stimulated with supernatant of RASF treated for 48h with methylprednisolone (urbason)
Platform GPL96: [HG-U133A] Affymetrix Human Genome U133A Array
Indicator
COX-2, CXCR-4, IL-1RN, IL-6/8, MMP-10/12, TLR-2, ADORA2A, BCL2-A1, CTGF, CXCR-7, CYR-61, HSD11B-1, IL-23A, MARCKS, MXRA-5, NDUFA4L2, NR4A3, SMS, STS, TNFAIP-2
TXNIP
Download Download data in GEO database