Research article

Antirheumatic drug response signatures in human chondrocytes: potential molecular targets to stimulate cartilage regeneration

Kristin Andreas¹ , Thomas Häupl² , Carsten Lübke³ , Jochen Ringe¹ , Lars Morawietz⁴ , Anja Wachtel¹ , Michael Sittinger¹ and Christian Kaps⁵

¹  Tissue Engineering Laboratory and Berlin – Brandenburg Center for Regenerative Therapies, Department of Rheumatology, Charité – Universitätsmedizin Berlin, Tucholskystrasse 2, 10117 Berlin, Germany

²  Tissue Engineering Laboratory, Department of Rheumatology, Charité – Universitätsmedizin Berlin, Tucholskystrasse 2, 10117 Berlin, Germany

³  University of Applied Sciences Wildau, Biosystems Technology, Bahnhofstrasse 1, 15745 Wildau, Germany

⁴  Institute of Pathology, Charité – Universitätsmedizin Berlin, Charitéplatz 1, 10117 Berlin, Germany

⁵  TransTissueTechnologies GmbH, Tucholskystrasse 2, 10117 Berlin, Germany

author email corresponding author email

Arthritis Research & Therapy 2009, 11:R15doi:10.1186/ar2605

Published:	3 February 2009

Abstract

Introduction

Rheumatoid arthritis (RA) leads to progressive destruction of articular cartilage. This study aimed to disclose major mechanisms of antirheumatic drug action on human chondrocytes and to reveal marker and pharmacological target genes that are involved in cartilage dysfunction and regeneration.

Methods

An interactive in vitro cultivation system composed of human chondrocyte alginate cultures and conditioned supernatant of SV40 T-antigen immortalised human synovial fibroblasts was used. Chondrocyte alginate cultures were stimulated with supernatant of RA synovial fibroblasts, of healthy donor synovial fibroblasts, and of RA synovial fibroblasts that have been antirheumatically treated with disease-modifying antirheumatic drugs (DMARDs) (azathioprine, gold sodium thiomalate, chloroquine phosphate, and methotrexate), nonsteroidal anti-inflammatory drugs (NSAIDs) (piroxicam and diclofenac), or steroidal anti-inflammatory drugs (SAIDs) (methylprednisolone and prednisolone). Chondrocyte gene expression profile was analysed using microarrays. Real-time reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay were performed for validation of microarray data.

Results

Genome-wide expression analysis revealed 110 RA-related genes in human chondrocytes: expression of catabolic mediators (inflammation, cytokines/chemokines, and matrix degradation) was induced, and expression of anabolic mediators (matrix synthesis and proliferation/differentiation) was repressed. Potential marker genes to define and influence cartilage/chondrocyte integrity and regeneration were determined and include already established genes (COX-2, CXCR-4, IL-1RN, IL-6/8, MMP-10/12, and TLR-2) and novel genes (ADORA2A, BCL2-A1, CTGF, CXCR-7, CYR-61, HSD11B-1, IL-23A, MARCKS, MXRA-5, NDUFA4L2, NR4A3, SMS, STS, TNFAIP-2, and TXNIP). Antirheumatic treatment with SAIDs showed complete and strong reversion of RA-related gene expression in human chondrocytes, whereas treatment with NSAIDs and the DMARD chloroquine phosphate had only moderate to minor effects. Treatment with the DMARDs azathioprine, gold sodium thiomalate, and methotrexate efficiently reverted chondrocyte RA-related gene expression toward the 'healthy' level. Pathways of cytokine-cytokine receptor interaction, transforming growth factor-beta/Toll-like receptor/Jak-STAT (signal transducer and activator of transcription) signalling and extracellular matrix receptor interaction were targeted by antirheumatics.

Conclusions

Our findings indicate that RA-relevant stimuli result in the molecular activation of catabolic and inflammatory processes in human chondrocytes that are reverted by antirheumatic treatment. Candidate genes that evolved in this study for new therapeutic approaches include suppression of specific immune responses (COX-2, IL-23A, and IL-6) and activation of cartilage regeneration (CTGF and CYR-61).