Combination of subtherapeutic anti-TNF dose with dasatinib restores clinical and molecular arthritogenic profiles better than standard anti-TNF treatment
New medications for Rheumatoid Arthritis (RA) have emerged in the last decades, including Disease Modifying Antirheumatic Drugs (DMARDs) and biologics. However, there is no known cure, since a significant proportion of patients remain or become non-responders to current therapies. The development of new mode-of-action treatment schemes involving combination therapies could prove successful for the treatment of a greater number of RA patients. In this work we investigated the effect of the Tyrosine Kinase inhibitors (TKIs) dasatinib and bosutinib, on the human TNF-dependent Tg197 arthritis mouse model. The inhibitors were administered either as a monotherapy or in combination with a subtherapeutic dose of anti-hTNF biologics and their therapeutic effect was assessed clinically, histopathologically as well as via gene expression analysis and was compared to that of an efficient TNF monotherapy. We found that dasatinib and, to a lesser extent, bosutinib inhibited the production of TNF and proinflammatory chemokines from arthritogenic synovial fibroblasts. Dasatinib, but not bosutinib, also ameliorated significantly and in a dose-depended manner both the clinical and histopathological signs of Tg197 arthritis. Combination of dasatinib with a subtherapeutic dose of anti-hTNF biologic agents, resulted in a synergistic inhibitory effect abolishing all arthritis symptoms. Gene expression analysis of whole joint tissue of Tg197 mice revealed that the combination of dasatinib with a low subtherapeutic dose of Infliximab most efficiently restores the pathogenic gene expression profile to that of the healthy state compared to either treatment administered as a monotherapy. Our findings show that dasatinib exhibits a therapeutic effect in TNF-driven arthritis and can act in synergy with a subtherapeutic anti-hTNF dose to effectively treat the clinical and histopathological signs of the pathology. The combination of dasatinib and anti-hTNF exhibits a distinct mode of action in restoring the arthritogenic gene signature to that of a healthy profile. Potential clinical applications of combination therapies with kinase inhibitors and anti-TNF agents, may provide an interesting alternative to high-dose anti-hTNF monotherapy and increase the number of patients responding to treatment. Files in the this dataset contain the total normalized expression values for all replicates of both untreated (wild-type and diseased) and treated (infliximab, dasatinib and dasatinib+infliximab) animal samples.
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Total RNA was isolated from whole joint tissues of 3 mice from each experimental treatment group, using Trizol reagent in combination with the RNeasy mini kit (Qiagen), according to the manufacturer’s guidelines. All samples were hybridized on the Affymetrix GeneChip Mouse Gene 2.0 ST array. Data analysis, identification of differentially expressed genes, comparison and clustering of differential expression profiles and assessment of treatment efficiency at transcriptional level were performed as previously described (https://journals.plos.org/ploscompbiol/article/comments?id=10.1371/journal.pcbi.1006933). Normalized gene expression values as well as differential expression values (logFC) for the complete set of analyzed genes are provided in Additional File 2 and Additional File 3, respectively.