T. Okano

Knockout of endothelin type B receptor signaling attenuates bleomycin-induced skin sclerosis in mice

BackgroundEndothelin-1 (ET-1) is important in the pathogenesis of systemic sclerosis (SSc). ET-1 binds two receptors, endothelin type A (ETA) and endothelin type B (ETB). Dual ETA/ETB receptor antagonists and a selective ETA receptor antagonist are used clinically to treat SSc, and the effect of these antagonists on fibroblast activation has been described. However, the role of ETB receptor signaling in fibrogenesis is less clear. This study was conducted to evaluate the profibrotic function of ETB receptor signaling in a murine model of bleomycin (BLM)-induced scleroderma.MethodsWe used ETB receptor–knockout (ETBKO) mice, which are genetically rescued from lethal intestinal aganglionosis by an ETB receptor transgene driven by the human dopamine β-hydroxylase (DβH)-gene promoter, and wild-type mice with DβH-ETB (WT). BLM or phosphate-buffered saline (PBS) was administered subcutaneously by osmotic minipump, and skin fibrosis was assessed by dermal thickness, subcutaneous fat atrophy, and myofibroblast count in the dermis. Dermal fibroblasts isolated from ETBKO and WT mice were cultured in vitro, stimulated with BLM or ET-1, and the expression of profibrotic genes was compared by quantitative PCR.ResultsDermal thickness, subcutaneous fat atrophy, and myofibroblast counts in the dermis were significantly reduced in ETBKO mice compared to WT mice, after BLM treatment. Compared with wild-type, dermal fibroblasts isolated from ETBKO mice showed lower gene expressions of α-smooth muscle actin and collagen 1α1 in response to BLM or ET-1 stimulation in vitro.ConclusionsET-1–ETB receptor signaling is involved in skin sclerosis and in collagen synthesis by dermal fibroblasts.

FRI0553 The efficacy of 2-years denosumab treatment for glucocorticoid-induced osteoporosis (GIOP)

Background Osteoporosis is one of the important adverse effects in the glucocorticoids treatment for the patients with rheumatoid arthritis (RA) and connective tissue diseases (CTDs). Although the usefulness of denosmab for primary osteoporosis has been well-established, the efficacy for GIOP remains unclear. Objectives This study aimed to clarify the therapeutic effects of denosumab for GIOP. Methods We evaluated bone mineral density (BMD) and serum markers of bone metabolism (BAP, NTx, TRACP-5b and P1NP) of patients who had been treated with over 5mg of predonisolone for RA and CTDs, and denosumab for GIOP, for two years in Kobe University Hospital. BMD and serum markers were evaluated every six months for 2 years from the baseline. The changes of those data from baseline were analyzed by Students t test using GraphPad Prism 5 software and p<0.05 was considered statistically significant. Results Number of the patients were 53 (male: 4 cases, female: 49 cases), and their characteristics at the beginning of denosumab treatment were as below; age: 64.19±12.0 years old, dose of prednisolon: 10.59±9.97mg/day, BMD of lumber spine: 0.768±0.112g/cm3, T-score of lumber spine: -2.28±1.01, BMD of femoral neck: 0.540±0.085g/cm3, T-score of femoral neck: -2.28±0.76. After 2-years denosumab treatment, T-scores of lumber spine (0.54±0.39 gain) and femoral neck (0.13±0.26 gain) were significantly increased from baseline (Figure; mean ± SEM. *:p<0.05). In addition, the serum markers of bone metabolism, both absorption and formation, were significantly suppressed with denosumab. Conclusions Denosumab can suppress bone metabolic turnover, and increase lumber spine and femoral neck T-scores of GIOP patients. Disclosure of Interest None declared

THU0611 Subjective Well-Being of Japanese RA Patients Who Reach Treatment Target Is Higher than The Japanese Average

Background Subjective Well-Being (SWB) refers to a construct which includes ones emotional responses, domain satisfactions, and global judgments of life satisfaction. Many variables such as age, sex, income, employment and marital status are related to SWB. Being healthy is also an important factor of SWB. RA is a chronic illness which is considered to influence SWB. However, little is known about the impact of RA on SWB. Objectives To evaluate the SWB of Japanese RA patients and identify factors that associate with SWB. Methods This study was done in cooperation with the Cabinet Office Government of Japan Economic and Social Research Institute. This institute had previously conducted the “Well-being studies 2014” surveying the SWB of randomly selected Japanese citizens. In this survey, SWB was determined by having the participants rate their happiness between 10 (Very happy) and 0 (very unhappy). The Well-being studies 2014 also included a questionnaire consisting of 56 questions covering topics closely associated with well-being such as socioeconomic and health status. The same survey was done for RA patients at Kobe University Hospital and clinical data including disease duration, stage, class, disease activity, HAQ, complications and the therapeutic drug was also collected at the same time. Results Multivariate analysis on data including RA patients (n=339) and Japanese controls (n=7690) revealed that RA patients with high or moderate disease activity had similar SWB scores as Japanese controls. However, the SWB of RA patients with remission or low disease activity were higher than Japanese controls. Age, sex, marital status, presence of child, household income, financial leeway, working status, psychological distress (Kessler 6 scale), self-assessment of health, and social connection were all associated with SWB. Conclusions Japanese RA patients may be able to get higher SWB than Japanese controls by achieving treatment target. Disclosure of Interest None declared

OP0150 3-Bromopyruvate Ameliorates Autoimmune Arthritis by Exerting A Dual Effect on Both Th17 and Treg Cell Differentiation and Dendritic Cell Activation

Background Recent studies have shown that cellular metabolism plays an important role in regulating immune cell function. In the process of cell differentiation, both interleukin-17-producing T (Th17) cells and dendritic cells (DCs) show increase of glycolytic activity by upregulating glycolytic enzymes, such as hexokinase-II (HK-II). Blocking glycolysis by 2-deoxyglucose has recently been demonstrated to inhibit Th17 cell differentiation while promote regulatory T (Treg) cell generation, and then ameliorate experimental autoimmune encephalitis model. Objectives The aim of this study is to verify the effect of 3-bromopyruvate (BrPA), a specific inhibitor of HK-II, on the differentiation and function of immune cells and on experimental arthritis in SKG mice. Methods Arthritis was induced in SKG mice by Zymosan A injection. BrPA (5 mg/kg) was administered subcutaneously once daily. CD4+ T cells from splenocytes of unimmunized SKG mice were cultured with anti-CD3/anti-CD28, anti-IFN-γ, anti-IL-4, IL-6, TGF-β, and IL-2, with or without BrPA for 5 days, and then analyzed by flow cytometry. Bone marrow (BM) cells from unimmunized SKG mice were cultured with GM-CSF and IL-4 (for 3 days), and with LPS (for 1 day) with or without BrPA, and then analyzed by flow cytometry. Results Treatment with BrPA significantly ameliorated arthritis of SKG mice (Figure). Histological scores of arthritis in BrPA-treated mice were significantly reduced compared with those in control mice. Flow cytometric analysis revealed that significant increase of Treg cells, decrease of Th17 cells, and decrease of CD40+CD86+CD11b+CD11c+ activated DCs were observed in the splenocytes from BrPA-treated mice. BrPA did not affect IFN-γ-producing Th1 cells nor IL-4-producing Th2 cells. In vitro, BrPA facilitated the differentiation of Treg cells, while it inhibited the development of Th17 cells. In addition, treatment of BM cells with BrPA inhibited the differentiation of activated DCs and the production of TNF-α and IL-6 from DCs. Conclusions BrPA facilitates the differentiation of Treg cells, and inhibits the development of Th17 cells and the activation of DCs, and ameliorates arthritis in the SKG mice. Disclosure of Interest None declared

AB0192 Some of The Painful RA Patients Underrate Global Health VAS at Hospitals

Background Evaluating patient global VAS is one of the most essential process in RA practice. Despite reliability of patient global VAS being highly important in clinical practice, there has been no study comparing global VAS scores obtained at hospitals and those obtained at home where patients answer anonymously. Objectives To compare the patient global VAS obtained before clinical examination in hospital with those answered anonymously at home. Methods We asked RA patients to answer and mail the EQ5D data set anonymously. EQ5D consisted of 5 component questions (mobility, self-care, usual activities, pain/discomfort, anxiety/depression) and patient assessment global health VAS. EQ5D global VAS is anonymized patient global VAS evaluated at home. We compared the patient global VAS which is routinely surveyed at hospital before clinical examination with those surveyed anonymously at home. Results The anonymized VAS score was higher than those routinely evaluated at hospital (p<0.0001). Global VAS scores obtained at hospital poorly correlated with those obtained anonymously at home (r=0.426). We compared patients who had higher patient global VAS at hospital than anonymized VAS at home with patients who had lower patient global VAS at hospital than anonymized VAS at home. Pain VAS scores remained to be risk factor to be higher anonymized VAS at home than those routinely evaluated at hospital after multivariate analysis. Conclusions Discrepancy exists between patient global VAS evaluated in the hospital before clinical examination and those evaluated anonymously at home. There is a possibility that patients rating high pain VAS are underrating their global VAS scores at hospital. Disclosure of Interest None declared

OP0297 Knockout of Endothelin Type B Receptor Signaling Attenuates Bleomycin-Induced Skin Sclerosis in Mice

Background Endothelin-1 (ET-1) is important in the pathogenesis of systemic sclerosis (SSc). It has been reported that plasma concentration of ET-1 are higher in SSc patients, and that endothelin receptor expression are also increased in lungs and skins in the patients. ET-1 binds two receptors, endothelin type A (ETA) and endothelin type B (ETB). Dual ETA/ETB receptor antagonists and a selective ETA receptor antagonist are used clinically to treat SSc, and the effect of these antagonists on fibroblast activation has been described. However, the role of ETB receptor signaling in fibrogenesis is less clear. Objectives This study was conducted to evaluate the profibrotic function of ETB receptor signaling in a murine model of bleomycin (BLM)-induced scleroderma. Methods We used ETB receptor–knockout (ETBKO) mice, which are genetically rescued from lethal intestinal aganglionosis by an ETB receptor transgene driven by the human dopamine β-hydroxylase (DβH)-gene promoter, and wild-type mice with DβH-ETB (WT). BLM or phosphate-buffered saline (PBS) was administered subcutaneously by osmotic minipump, and skin fibrosis was assessed by dermal thickness, subcutaneous fat atrophy, and alpha-smooth muscle actin (αSMA)-expressing myofibroblast count in the dermis. Dermal fibroblasts isolated from ETBKO and WT mice were cultured in vitro, stimulated with BLM or ET-1, and the expression of profibrotic genes was compared by quantitative PCR. Results Dermal thickness, subcutaneous fat atrophy, and myofibroblast counts in the dermis were significantly reduced in ETBKO mice compared to WT mice, after BLM treatment. Compared with wild-type, dermal fibroblasts isolated from ETBKO mice showed lower gene expressions of αSMA and collagen 1α1 in response to BLM or ET-1 stimulation in vitro. Conclusions ET-1–ETB receptor signaling is involved in skin sclerosis and in collagen synthesis by dermal fibroblasts. Disclosure of Interest None declared