Targeting Activated Synovial Fibroblasts in Rheumatoid Arthritis by Peficitinib

Background: Synovial fibroblasts (SF) play a major role in the pathogenesis of rheumatoid arthritis (RA) and develop an aggressive phenotype destroying cartilage and bone, thus termed RASF. JAK inhibitors have shown to be an efficient therapeutic option in RA treatment, but less is known about the effect of JAK inhibitors on activated RASF. The aim of the study was to examine the effects of JAK inhibitors on activated RASF. Methods: Synovium of RA patients was obtained during knee replacement surgeries. Synoviocytes were isolated and pretreated with JAK inhibitors. Pro-inflammatory cytokines and matrix degrading proteinases were measured by ELISA in supernatant after stimulation with oncostatin M or IL-1β. The proliferation of RASF was measured by BrdU incorporation. Cell culture inserts were used to evaluate cell migration. For adhesion assays, RASF were seeded in culture plates. Then, plates were extensively shaken and adherent RASF quantified. Cell viability, cytotoxicity and apoptosis were measured using the ApoTox-Glo™ Triplex and the CellTox™ Green Cytotoxicity Assay. Results: Tofacitinib and baricitinib decreased the IL-6 release of RASF stimulated with oncostatin M. JAK inhibition attenuated the IL-6 release of IL-1β activated and with soluble IL-6 receptor treated RASF. In contrast, only peficitinib and filgotinib decreased the IL-6 release of RASF activated with IL-1β. Peficitinib decreased also the MMP-3, CXCL8, and CXCL1 release at 5 μM. Moreover, peficitinib was the only JAK inhibitor suppressing proliferation of activated RASF at 1 μM. Peficitinib further decreased the migration of RASF without being cytotoxic or pro-apoptotic and without altering cell adhesion. Conclusions: JAK inhibitors effectively suppress the inflammatory response induced by oncostatin M and by transsignaling of IL-6 in RASF. Only peficitinib modulated the IL-1β-induced response of RASF and their proliferation in vitro at concentrations close to reported Cmax values of well tolerated doses in vivo. In contrast to filgotinib, peficitinib also highly suppressed RASF migration showing the potential of peficitinib to target RASF.

[1]  D. Veale,et al.  JAK/STAT Blockade Alters Synovial Bioenergetics, Mitochondrial Function, and Proinflammatory Mediators in Rheumatoid Arthritis , 2018, Arthritis & rheumatology.

[2]  A. Van der Aa,et al.  Efficacy, Safety, Pharmacokinetics, and Pharmacodynamics of Filgotinib, a Selective JAK‐1 Inhibitor, After Short‐Term Treatment of Rheumatoid Arthritis: Results of Two Randomized Phase IIa Trials , 2017, Arthritis & rheumatology.

[3]  A. Araujo,et al.  Prevalence of rheumatoid arthritis in the United States adult population in healthcare claims databases, 2004–2014 , 2017, Rheumatology International.

[4]  Tsutomu Takeuchi,et al.  EULAR recommendations for the management of rheumatoid arthritis with synthetic and biological disease-modifying antirheumatic drugs: 2016 update , 2010, Annals of the rheumatic diseases.

[5]  Takayuki Inoue,et al.  A novel JAK inhibitor, peficitinib, demonstrates potent efficacy in a rat adjuvant-induced arthritis model. , 2017, Journal of pharmacological sciences.

[6]  J. Bertrand,et al.  Stable activation of fibroblasts in rheumatic arthritis-causes and consequences. , 2016, Rheumatology.

[7]  S. Swan,et al.  Pharmacokinetics, Pharmacodynamics, and Safety of ASP015K (Peficitinib), a New Janus Kinase Inhibitor, in Healthy Subjects , 2016, Clinical pharmacology in drug development.

[8]  J. Kushner,et al.  Extended‐Release Once‐Daily Formulation of Tofacitinib: Evaluation of Pharmacokinetics Compared With Immediate‐Release Tofacitinib and Impact of Food , 2016, Journal of clinical pharmacology.

[9]  Massimo Gadina,et al.  Type I/II cytokines, JAKs, and new strategies for treating autoimmune diseases , 2016, Nature Reviews Rheumatology.

[10]  P. Scherle,et al.  The pharmacokinetics, pharmacodynamics, and safety of baricitinib, an oral JAK 1/2 inhibitor, in healthy volunteers , 2014, Journal of clinical pharmacology.

[11]  K. Migita,et al.  Inhibition of Janus kinase/signal transducer and activator of transcription (JAK/STAT) signalling pathway in rheumatoid synovial fibroblasts using small molecule compounds , 2013, Clinical and experimental immunology.

[12]  L. Nelles,et al.  Preclinical Characterization of GLPG0634, a Selective Inhibitor of JAK1, for the Treatment of Inflammatory Diseases , 2013, The Journal of Immunology.

[13]  Yoshiya Tanaka,et al.  The JAK inhibitor tofacitinib regulates synovitis through inhibition of interferon-γ and interleukin-17 production by human CD4+ T cells. , 2012, Arthritis and rheumatism.

[14]  T. Shimamura,et al.  Interleukin‐6 upregulates expression of ADAMTS‐4 in fibroblast‐like synoviocytes from patients with rheumatoid arthritis , 2012, International journal of rheumatic diseases.

[15]  G. Firestein,et al.  The JAK inhibitor CP-690,550 (tofacitinib) inhibits TNF-induced chemokine expression in fibroblast-like synoviocytes: autocrine role of type I interferon , 2011, Annals of the rheumatic diseases.

[16]  K. Migita,et al.  CP690,550 inhibits oncostatin M-induced JAK/STAT signaling pathway in rheumatoid synoviocytes , 2011, Arthritis research & therapy.

[17]  U. Müller-Ladner,et al.  Cell culture and passaging alters gene expression pattern and proliferation rate in rheumatoid arthritis synovial fibroblasts , 2010, Arthritis research & therapy.

[18]  P. Robbins,et al.  Synovial fibroblasts spread rheumatoid arthritis to unaffected joints , 2009, Nature Medicine.

[19]  M. Hashizume,et al.  IL-6 trans-signalling directly induces RANKL on fibroblast-like synovial cells and is involved in RANKL induction by TNF-alpha and IL-17. , 2008, Rheumatology.

[20]  P. Robbins,et al.  Inhibition of cartilage destruction by double gene transfer of IL-1Ra and IL-10 involves the activin pathway , 2002, Gene Therapy.

[21]  K. Miyazawa,et al.  Regulation of Interleukin-1β-induced Interleukin-6 Gene Expression in Human Fibroblast-like Synoviocytes by p38 Mitogen-activated Protein Kinase* , 1998, The Journal of Biological Chemistry.

[22]  K. Miyazawa,et al.  Transcriptional Roles of CCAAT/Enhancer Binding Protein-β, Nuclear Factor-κB, and C-promoter Binding Factor 1 in Interleukin (IL)-1β-induced IL-6 Synthesis by Human Rheumatoid Fibroblast-like Synoviocytes* , 1998, The Journal of Biological Chemistry.

[23]  E. Martı́n-Mola,et al.  Soluble interleukin 6 (IL-6) receptor and IL-6 levels in serum and synovial fluid of patients with different arthropathies. , 1997, The Journal of rheumatology.

[24]  T. Vischer,et al.  Concentrations and origins of soluble interleukin 6 receptor-alpha in serum and synovial fluid. , 1997, The Journal of rheumatology.

[25]  M. Liang,et al.  The American Rheumatism Association 1987 revised criteria for the classification of rheumatoid arthritis. , 1988, Arthritis and rheumatism.