Gene Expression Profiling of Rheumatoid Arthritis Synovial Cells Treated with Antirheumatic Drugs

Abstract

Nonbiological therapeutics are frequently used for the treatment of patients with rheumatoid arthritis (RA). Because the mechanisms of action of these therapeutics are unclear, the authors aimed to elucidate the molecular effects of typical antirheumatic drugs on the expression profile of RA-related genes expressed in activated synovial fibroblasts. For reasons of standardization and comparability, immortalized synovial fibroblasts derived from RA (RASF) and normal donors (NDSF) were treated with methotrexate, prednisolone, or diclofenac and used for gene expression profiling with oligonucleotide microarrays. The cytotoxicity of the antirheumatic drugs was tested in different concentrations by MTS tetrazolium assay. Genes that were differentially expressed in RASF compared to NDSF and reverted by treatment with antirheumatic drugs were verified by semiquantitative polymerase chain reaction and by chemiluminescent enzyme immunoassay. Treatment with methotrexate resulted in the reversion of the RA-related expression profile of genes associated with growth and apoptosis including insulin-like growth factor binding protein 3, retinoic acid induced 3, and caveolin 2 as well as in the re-expression of the cell adhesion molecule integrin α6. Prednisolone reverted the RA-related profile of genes that are known from inflammation and suppressed interleukins 1β and 8. Low or high doses of diclofenac had no effect on the expression profile of genes related to RA in synovial fibroblasts. These data give the first insight into the mechanisms of action of common antirheumatic drugs used for the treatment of arthritides. Synovial fibroblasts reflect the disease-related pathophysiology and are useful tools for screening putative antirheumatic compounds. (Journal of Biomolecular Screening 2007:328-340)

Priming integrin α5 promotes human mesenchymal stromal cell osteoblast differentiation and osteogenesis

“…Subconfluent hMSCs were incubated with or without 10 Ϫ7 M dexamethasone for 1, 3, or 7 days, total RNA was isolated, and gene expression profiling with Affymetrix HG-U133 Plus 2.0 arrays was performed as described in ref. 21. Raw gene expression data were processed with the GCOS 1.2 software for signal calculation, and comparison analysis and cellular pathways were analyzed using statistical tools.…”

“…S1 A-D). We analyzed the gene expression profile in these differentiating hMSCs using HG-U133 Plus 2.0 arrays consisting of 54,675 probe sets (21). Analysis of normalized data from three different donors revealed that ITGA5 expression was increased Ͼ2-fold in dexamethasone-treated hMSCs compared with that in untreated cells.…”

Differential Gene Expression Profiles May Differentiate Responder and Nonresponder Patients with Rheumatoid Arthritis for Methotrexate (MTX) Monotherapy and MTX plus Tumor Necrosis Factor Inhibitor Combined Therapy

“…The only study available in the literature evaluated the effect of in vitro MTX treatment on RA and on control synovial fibroblasts, reporting that MTX reversed the modulated expression of genes related to apoptosis and cell adhesion in RA fibroblasts to the levels observed in control fibroblasts 28 . In our series, despite the small number of patients, we clearly observed modulation of genes implicated in apoptosis in MTX nonresponder patients when compared to responders.…”

Association of Blood Gene Expressions in Rheumatoid Arthritis Patients With Clinical and Laboratory Parameters Before and After Methotrexate Therapy

“…Напротив, рост числа эрозий и величины сужения су-ставной щели (ССЩ) у серопозитивных больных РА в хо-де терапии МТ сопровождался увеличением экспрессии ММП9 и катепсина К в крови [16,17]. Кроме того, норма-лизация экспрессии генов, ассоциированных с ростом клеток и апоптозной активностью, также ранее наблюда-лась в присутствии МТ в культуре синовиальных фиб-робластов in vitro [18].…”