Objective: Systemic sclerosis (SSc) is an autoimmune multisystem disease with poorly understood pathogenesis and ineffective treatment options. Soluble CD13 (sCD13), generated by the cleavage of cell surface CD13 via matrix metalloproteinase 14 (MMP14), signals through the bradykinin receptor B1 (B1R) to elicit pro‐inflammatory, pro‐arthritic, and pro‐angiogenic responses. In this study, we explored the antifibrotic potential of targeting the sCD13‐B1R axis in SSc.Methods: The expression of CD13, B1R, and MMP14 was examined in SSc skin and explanted dermal fibroblasts. The efficacy of B1R antagonists in the inhibition on fibrosis was determined in vitro and in vivo.Results: Expression of the genes for CD13, B1R, and MMP14 was elevated in skin biopsies from patients with diffuse cutaneous (dc) SSc. Notably, single‐cell analysis of SSc skin biopsies revealed the highest BDKRB1 expression in COL8A1‐positive myofibroblasts, a population exclusively seen in SSc. Transforming growth factor beta (TGFβ) induced the expression of BDKRB1 and production of sCD13 by dcSSc skin fibroblasts. Treatment of dcSSc fibroblasts with sCD13 promoted fibrotic gene expression, signaling, cell proliferation, migration, and gel contraction. The pro‐fibrotic responses of sCD13 or TGFβ were prevented by a B1R antagonist. Mice lacking Cd13 or Bdkrb1 were resistant to bleomycin‐induced skin fibrosis and inflammation. Pharmacological B1R inhibition had a comparable antifibrotic effect.Conclusion: These results are the first to demonstrate a key role for sCD13 in SSc skin fibrosis and suggest that targeting the sCD13‐B1R signaling axis is a promising novel therapeutic approach for SSc. image

Background: This study was performed to examine the effects of the Janus kinase (JAK) inhibitor peficitinib on fibroblast-like synoviocytes (FLS) obtained from patients with rheumatoid arthritis (RA). Methods: To examine the expression of JAK1, JAK2, and JAK3 in RA synovial tissue (ST) and FLS, immunohistochemistry was performed. We investigated the effects of peficitinib on interleukin 6 and IL-6 receptor responses in RA FLS. Phosphorylation of STAT was determined by western blot. To examine the functional analysis of peficitinib, we performed a proliferation and chemotaxis assays with FLS using THP-1 and peripheral blood mononuclear cells (PBMC). The inflammatory mediator expression of FLS was estimated by enzyme-linked immunosorbent assay. Results: JAK1, JAK2, and JAK3 were expressed in RA STs and FLS. Phosphorylation of STAT1, STAT3, and STAT5 in RA FLS was suppressed by peficitinib in a concentration-dependent manner. Peficitinib-treated RA FLS-conditioned medium reduced THP-1 and PBMC migration (p < 0.05) and proliferation of RA FLS (p < 0.05). Peficitinib suppressed the secretion of MCP-1/CCL2 in the RA FLS supernatant (p < 0.05). Conclusion: Peficitinib suppressed the JAK-STAT pathway in RA FLS and also suppressed monocyte chemotaxis and proliferation of FLS through inhibition of inflammatory cytokines.

We examined whether the age of patients with rheumatoid arthritis was associated with adverse events (AEs) caused by biologic disease-modifying antirheumatic drugs (bDMARDs). Patients with rheumatoid arthritis using bDMARDs from Showa University Hospital, Showa University Northern Yokohama Hospital, and Showa University Koto Toyosu Hospital from January 2005 to December 2017 were eligible for this retrospective cohort study. The maximum observation period was determined to be 1 year. Outcomes in patients older and younger than 75 years were compared. The primary outcome was the rate of drug discontinuation because of AEs caused by bDMARDs. Univariate and multivariate analyses were performed using Pearson's chi-squared test and logistic regression analysis, respectively. A total of 416 patients were enrolled; median (interquartile range [IQR]): 60.0 (44.3 - 71.0) years and 84.6% women; patients ≥ 75 years were 67/416 (16.1%). The rates of drug discontinuation because of AEs caused by bDMARDs were 10.5% (7/67) in patients 75 years and older and 10.9% (38/349) in those younger than 75 years (relative risk 0.95, 95% confidential interval 0.45-2.24). In logistic regression analysis adjusted for covariates, the rate of drug discontinuation showed no significant difference between the patients ≥ 75 years and the those < 75 years (adjusted odds ratio 0.70, 95% confidential interval 0.29-1.75, P = .45). The rate of drug discontinuation because of AEs caused by bDMARDs was not significantly different between patients 75 years and older and patients younger than 75 years.

Background: Eosinophilic fasciitis (EF) is a rare disease that causes inflammation and fibrosis mainly in the fascia of the extremities with eosinophilia. It has been reported that the hypertrophied fascia in EF shows inflammatory cell infiltration by the lymphocytes and eosinophils and increased expression of fibrosis-related cytokines genes in fibroblast [1]. However, its pathophysiology in the fascia remains unresolved. Objectives: Therefore, we focused on fascial fibroblasts and aimed to determine the role of interleukin-4 (IL-4) in eosinophil and helper T cell infiltration and fibrosis in fascial fibroblast in EF. Methods: Fascial fibroblasts were obtained from fascia biopsy of a patient with EF, and were stimulated with pre- and post-treatment serum of a patient with EF and healthy control, followed by microarray to analyze gene expression. Fascial fibroblasts were stimulated with IL-4 10 ng/mL, and gene expression of IL-4 receptor and CCR3 ligands, CCL7 and CCL11 were measured by qPCR. Transforming growth factor (TGF) -β and periostin in the pre- and post-treatment serum of a patient with EF and conditioned medium of fascial fibroblasts stimulated with IL-4 were measured by ELISA. To examine the role of IL-4 in proliferation, we performed in proliferation assays using fascial fibroblasts treated with IL-4. CCR3-positive T cells in the fascial tissue of EF, dermatomyositis, and polymyositis patients were evaluated by immunostaining. Results: By microarray analysis, CCL7 and CCL11 expression of fascial fibroblasts stimulated with pre-treatment EF serum was higher than that in post-treatment EF serum and control serum. CCL7 and CCL11 mRNA in IL-4 stimulated facial fibroblasts were increased by 5.1-fold and 7.3-fold, respectively. TGF-β and periostin in IL-4 stimulated facial fibroblast conditioned medium were also increased. In addition, TGF-β and periostin in EF serum were gradually decreased by treatment for 4 and 10 weeks, compared to before treatment. Finally, fascial fibroblast proliferation was significantly increased by stimulation with IL-4. Furthermore, infiltration of CCR3-positive T cells was specific to the fascial tissue of EF. Conclusion: In EF, IL-4 enhances the production of CCR3 ligands, TGF-β, and periostin from fascial fibroblasts. As a result, it promotes the migration of eosinophils and CCR3-positive T helper cells to the fascia and fibrosis. These results suggest that inhibition of IL-4 pathway could be a novel strategy for eosinophilic fasciitis. References: [1]Igarashi A, Nashiro K, Kikuchi K, et al. Connective tissue growth factor gene expression in tissue sections from localized scleroderma, keloid, and other fibrotic skin disorders. J Invest Dermatol. 1996 Apr;106(4):729-33. Disclosure of Interests: None declared

Background: Hepatocyte growth factor (HGF), originally discovered as a mitogen of hepatocytes, binds to receptor-tyrosine kinase c-Met and has been shown to be a multi-functional cytokine that promotes processes such as cell proliferation, survival, differentiation, migration, and angiogenesis¹. Since HGF/c-Met signaling also leads to tumorigenesis and cancer invasion, that has recently attracted attention as a target for anticancer agents². However, in reports of rheumatoid arthritis (RA), though anti-inflammatory and antiangiogenic mechanisms related to HGF/c-Met signal inhibition have been reported, the role of HGF in RA bone destruction through monocyte migration remains unclear³. Objectives: To determine the expression of HGF in RA biological fluids, the role it plays in monocyte migration and the therapeutic effect of a savolitinib, a specific c-Met inhibitor, in arthritis model mice. Methods: HGF/c-Met expression in serum, synovial fluid (SF), and synovial tissues (STs) obtained from RA patients and control subjects, as well as RA fibroblast-like synoviocytes (FLSs) was evaluated by ELISA and immunostaining. To determine the function of HGF in RA SFs, we preincubated RA SFs with a neutralizing anti-HGF antibody and measured the ability of these SFs to induce the human acute monocytic leukemia cell line (THP-1) chemotaxis. Additionally, examinations of SKG mice treated with savolitinib (2.5 mg/kg/day) for 4 weeks were conducted. Results: HGF level in serum from RA patients was significantly higher as compared to the controls (930 ± 97 vs. 476 ± 97 pg/mL, p <0.01) and decreased by drug treatment for 24 weeks (1147 ± 284 vs. 539 ± 160 pg/mL, p <0.05). Additionally, HGF level in SF from RA patients was higher as compared to SF from osteoarthritis patients (1632 ± 366 vs. 566 ± 140 pg/mL, p <0.05). HGF and c-Met expressions were also noted in RA STs. Stimulation of RA-FLS with TNF-α increased HGF/c-Met expression in a concentration-dependent manner, and c-Met signal inhibition by SU11274 suppressed production of fractalkine/CX3CL1, CXCL16, and MIP-1α/CCL3 (mean 50%, 56%, 90%, respectively). When HGF was removed by immunoprecipitation, migration of THP-1 in RA-SF was suppressed (mean 23%). In SKG mice, savolitinib significantly suppressed ankle bone damage on µCT, with an associated reduction in number of tartrate-resistant acid phosphatase-positive osteoclasts. Conclusion: HGF is produced by inflammation in synovium associated with RA, and then activates monocyte migration to synovium tissue and promotes bone destruction through its own chemotactic effect as well as enhanced chemokine production. These results indicate that a strategy that targets c-Met signaling may be important for resolving bone destruction in RA. References: [1] Nakamura T, Nishizawa T, Hagiya M, Seki T, Shimonishi M, Sugimura A, Tashiro K, Shimizu S. Molecular cloning and expression of human hepatocyte growth factor. Nature. 1989 Nov 23;342(6248):440-3 [2] Lee D, Sung ES, Ahn JH, An S, Huh J, You WK. Development of antibody-based c-Met inhibitors for targeted cancer therapy. Immunotargets Ther. 2015 Feb 9;4:35-44. [3] Koch AE, Halloran MM, Hosaka S, Shah MR, Haskell CJ, Baker SK, Panos RJ, Haines GK, Bennett GL, Pope RM, Ferrara N. Hepatocyte growth factor. A cytokine mediating endothelial migration in inflammatory arthritis. Arthritis Rheum. 1996 Sep;39(9):1566-75 Disclosure of Interests: None declared