Mária Filková

Increased serum adiponectin levels in female patients with erosive compared with non-erosive osteoarthritis

Adipocytokines including adiponectin and resistin are suggested to be associated with obesity-related complications.1 In general, higher systemic concentrations of resistin and adiponectin compared with paired synovial fluid counterparts were demonstrated in patients with osteoarthritis of the knee.2–4 In contrast, resistin and adiponectin levels were found to be increased at local sites of inflammation in patients with rheumatoid arthritis.3–5 It is suggested that adiponectin in particular actively participates in the process of immune response, inflammation and matrix degradation in destructive arthritides.6 7nnErosive osteoarthritis represents a subtype of generalised osteoarthritis primarily affecting the small joints of the hands, with prominent local inflammation …

An update on biomarkers in axial spondyloarthritis.

Axial spondyloarthritis is a chronic inflammatory disease with the onset at a young age, and, if undiagnosed and untreated, it may result in permanent damage and lifelong disability. Rates of early diagnosis have improved, due in particular to the addition of magnetic resonance imaging into the diagnostic armamentaria; however, it is costly, not widely available, and requires experienced readers to interpret the findings. In addition to clinical measures and imaging techniques, biomarkers that will be described in this review may represent useful tools for diagnosis, monitoring disease activity and outcomes as well as therapeutic responses. Currently, HLA-B27 remains the best genetic biomarker for making a diagnosis, while CRP currently appears to be the best circulating measure for assessing disease activity, predicting structural progression and therapeutic response. Interestingly, key molecules in the pathogenesis of the disease and essential therapeutic targets, such as tumour necrosis factor (TNF)α, interleukin (IL)-17 and IL-23, show only limited association with disease characteristics or disease progression. Some genetic biomarkers and particularly anti-CD74 antibodies, may become a promising tool for the early diagnosis of axSpA. Further biomarkers, such as matrix metalloproteinases (MMP)-3, calprotectin (S100A8/9), vascular endothelial growth factor (VEGF), C-terminal telopeptide of type II collagen (CTX-II) or dickkopf-1 (DKK-1), are not sufficient to reflect disease activity, but may predict spinal structural progression. In addition, recent data have shown that monitoring calprotectin might represent a valuable biomarker of therapeutic response. However, all of these results need to be confirmed in large cohort studies prior to use in daily clinical practice.

Resistin in idiopathic inflammatory myopathies

IntroductionThe purpose of this study was to evaluate and compare the serum levels and local expression of resistin in patients with idiopathic inflammatory myopathies to controls, and to determine the relationship between resistin levels, inflammation and disease activity.MethodsSerum resistin levels were determined in 42 patients with inflammatory myopathies and 27 healthy controls. The association among resistin levels, inflammation, global disease activity and muscle strength was examined. The expression of resistin in muscle tissues from patients with inflammatory myopathies and healthy controls was evaluated. Gene expression and protein release from resistin-stimulated muscle and mononuclear cells were assessed.ResultsIn patients with inflammatory myopathies, the serum levels of resistin were significantly higher than those observed in controls (8.53 ± 6.84 vs. 4.54 ± 1.08 ng/ml, P < 0.0001) and correlated with C-reactive protein (CRP) levels (r = 0.328, P = 0.044) and myositis disease activity assessment visual analogue scales (MYOACT) (r = 0.382, P = 0.026). Stronger association was observed between the levels of serum resistin and CRP levels (r = 0.717, P = 0.037) as well as MYOACT (r = 0.798, P = 0.007), and there was a trend towards correlation between serum resistin and myoglobin levels (r = 0.650, P = 0.067) in anti-Jo-1 positive patients. Furthermore, in patients with dermatomyositis, serum resistin levels significantly correlated with MYOACT (r = 0.667, P = 0.001), creatine kinase (r = 0.739, P = 0.001) and myoglobin levels (r = 0.791, P = 0.0003) and showed a trend towards correlation with CRP levels (r = 0.447, P = 0.067). Resistin expression in muscle tissue was significantly higher in patients with inflammatory myopathies compared to controls, and resistin induced the expression of interleukins (IL)-1β and IL-6 and monocyte chemoattractant protein (MCP)-1 in mononuclear cells but not in myocytes.ConclusionsThe results of this study indicate that higher levels of serum resistin are associated with inflammation, higher global disease activity index and muscle injury in patients with myositis-specific anti-Jo-1 antibody and patients with dermatomyositis. Furthermore, up-regulation of resistin in muscle tissue and resistin-induced synthesis of pro-inflammatory cytokines in mononuclear cells suggest a potential role for resistin in the pathogenesis of inflammatory myopathies.

The Role of Resistin in Inflammatory Myopathies

Both immune and non-immune mechanisms are involved in muscle damage and dysfunction occurring in idiopathic inflammatory myopathies (IIMs). Crosstalk among inflammatory cells, muscle and endothelial cells is essential in the pathogenesis of IIMs. Resistin, originally described as an adipokine linking obesity and insulin resistance in rodents, has been shown a pro-inflammatory molecule in humans. Besides its direct effect on production of several inflammatory mediators, resistin influences chemotaxis, migration, proliferation, cell survival, endothelial dysfunction and metabolism—all aspects implicated in the pathogenesis of IIMs. Up-regulation of resistin in muscle tissue and elevated serum resistin levels have been recently demonstrated in patients with IIMs. In addition, serum levels of resistin reflected global disease activity, including extramuscular organ involvement, in patients with this disease. However, there are currently not sufficient data to distinguish the features of resistin that cause injury of muscle tissue from those that promote muscle regeneration and repair. The aim of this review is therefore to summarize current knowledge about potential implication of resistin in idiopathic inflammatory myopathies.

Protein Tyrosine Phosphatase Nonreceptor Type 2: An Important Regulator of lnterleukin‐6 Production in Rheumatoid Arthritis Synovial Fibroblasts

To investigate the role of protein tyrosine phosphatase nonreceptor type 2 (PTPN2) in the pathogenesis of rheumatoid arthritis (RA).

Interleukin-20 is triggered by TLR ligands and associates with disease activity in patients with rheumatoid arthritis

Background: Interleukin (IL)‐20 is a pro‐inflammatory cytokine that may be implicated in the pathogenesis of rheumatoid arthritis (RA). This study aimed to determine the association between IL‐20 and disease activity in patients with RA. Methods: The levels of serum and synovial fluid IL‐20 were measured in patients with RA and OA. The disease activity was assessed based on the Disease Activity Score of 28 joints (DAS28). The expression of IL‐20 in synovial tissue samples from patients with RA and OA were determined by immunohistochemistry. Immunofluorescence staining was used to co‐localize IL‐20 with selected cells. The secretion of IL‐20 was analysed in human peripheral blood mononuclear cells (PBMCs) of patients with RA. Results: Synovial fluid and synovial tissue IL‐20 were significantly increased in patients with RA compared with patients with OA. The expression of IL‐20 in RA synovial tissue was particularly associated with macrophages and neutrophil granulocytes, but also with synovial fibroblasts and lymphocytes. The IL‐20 levels in synovial fluid correlated with DAS28 (r = 0.434; p = 0.015) and were significantly elevated in anti‐CCP positive RA compared with anti‐CCP negative RA (122.3 ± 104.1 pg/ml and 45.9 ± 35.8 pg/ml; p = 0.008). IL‐20 production from PBMCs was induced by Poly I:C and LPS but not with pro‐inflammatory cytokines, such as TNF‐&agr; or IL‐1. Conclusion: Our data showed that IL‐20 is independently associated with RA disease activity and may be triggered by TLR ligands at local sites of inflammation. HighlightsElevated synovial fluid IL‐20 is associated with RA disease activity.IL‐20 levels are higher in anti‐CCP positive RA patients.IL‐20 production from PBMCs is induced by TNF‐independent TLR pathways.

SAT0099 Polypharmacy is associated with an increased risk of adverse outcomes in patients with rheumatoid arthritis

Background In the general population, polypharmacy (PP) is associated with increased risk of adverse events. The relationship between adverse outcomes and PP in Rheumatoid Arthritis (RA) has not been studied in depth. The mantra of treatment in RA encourages PP through combination Disease Modifying Anti-Rheumatic Drugs (DMARD). Objectives To study the relationship between PP and serious adverse events in RA, including the influence of DMARDs within the PP count. Methods Data from the British Society for Rheumatology Biologics Register were analysed. PP was defined as number of drugs co-prescribed at baseline, with two models: (1) including DMARDs (2) excluding DMARDs from the medication count. PP was stratified by 0–5, 6–9 and >10. Patients were studied from initiation of 1st biologic until 1st serious adverse event (SAE), 3 years of follow up, or last available visit, whichever came first. A Cox-proportional hazard model was used, with adjustment for a priori selected cofounders. Results This study included 15,004 patients commencing biologics. The demographics are shown in table 1. Excluding DMARDs from the PP cohort, 7,115 (47%) of the patients were taking up to 5 drugs; 6,010 (40%) were taking 6 to 9 drugs; 1,870 (12%) were taking 10 or more medications. Higher levels of PP associated with older age, more severe disease, and longer disease duration. PP predictably associated with comorbidities; the relationship was not linear: comorbidity count appeared to show a ceiling effect. The overall incidence of SAEs was 25.5/100 person years (95% CI 24.7–26.3). The rate of SAEs increased across the PP counts (See Table 1). The relationship remained significant after adjusting for comorbidities. Including DMARDs within the PP count attenuated the association.Table 1 All Patients 0–5 drugs 6–9 drugs >10 drugs PP count excluding DMARDs N=15,004 n=7,115 n=6,019 n=1,870 Baseline characteristics u2003Mean Age in years 56.3 54.0 57.6 61.0 u2003Mean DAS 28 (SD) 4.30 (1.76) 4.17 (1.79) 4.51 (1.67) 4.88 (1.67) u2003Mean HAQ (SD) 1.93 (0.64) 1.85 (0.65) 2.10 (0.56) 2.15 (0.58) u2003Mean Disease Duration (SD) in years 12.59 (9.72) 11.96 (9.32) 13.79 (10.26) 14.67 (10.96) u2003Comorbidity (SD) 1.87 (0.80) 1.65 (0.74) 2.29 (0.73) 2.57 (0.68) Analysis of Serious Adverse Events u2003Exposure time (person-years) 14,200 9,690 3,706 804 u2003Event count (single failure model) 3261 2002 1251 368 u2003Incidence rate (95% CI) 25.5 (24.7–26.3) 20.6 (19.7–21.5) 33.7 (31.9–35.6) 45.7 (41.3–50.7) u2003Including DMARDs in PP model u2003Unadjusted HR (95% CI) – Ref 1.20 (1.11–1.29) 1.82 (1.66–1.99) u2003Adjusted HR (95% CI) – Ref 1.05 (0.97–1.13) 1.39 (1.26–1.54) u2003Excluding DMARDs in PP model u2003Unadjusted HR (95% CI) – Ref 1.63 (1.52–1.75) 2.21 (1.98–2.47) u2003Adjusted HR (95% CI) – Ref 1.18 (1.09–1.28) 1.35 (1.19–1.53) Adjusted for age, sex, DAS, HAQ, disease duration and comorbidities. Conclusions PP is common in patients with RA and is associated with adverse outcomes especially when patients are on >10 drugs. Including or excluding DMARDs from the PP model had negligible impact on findings. The relationship between PP and comorbidity is worthy of further research, as PP represents a potentially simple but valuable predictor of adverse outcomes, and a suitable surrogate for comorbidity in epidemiological analyses. Disclosure of Interest None declared

A6.26 Analysis of circulating mirnas in patients with axial sponyloarthritis with different spinal involvement

Background and objectives The altered expression of miRNAs and dysregulation of their target genes has been shown to contribute to pathophysiology of many autoimmune diseases. In addition, circulating miRNAs may serve as diagnostic and/or prognostic biomarkers. Our aim was to identify circulating miRNAs in patients with axial spondyloarthritis (AxSpA) and to investigate their relationship with spinal involvement and disease activity. Material and methods Total RNA was isolated using phenol-chloroform extraction from plasma of 20 patients with non-radiographic AxSpA (nr-AxSpA), 48 patients with radiographic AxSpA (with and without spinal involvement, including 6 patients with a bamboo spine), and 29 healthy controls (HC). A comprehensive analysis of miRNAs was performed using TaqMan® Low Density Array (TLDA) in 5 samples from each group. Expression of miRNAs was further confirmed by single assays in the remaining samples and the levels were normalised to C. elegans spike-in controls. Disease activity was assessed based on C-reactive protein (CRP) and Bath Ankylosing Spondylitis Disease Activity Index (BASDAI). Data were analysed using one-way ANOVA, unpaired t-test with Welch’s correction and Pearson correlation. Results Out of 760 miRNAs analysed by TLDA, 21 miRNAs were selected for further analysis, according to differences in the expression and role in the inflammation and bone metabolism. Fourteen miRNAs were significantly lower (p < 0.05) in all patients with AxSpA compared to HC. In all patients with radiographic AxSpA (r-AxSpA), 20 miRNAs were significantly lower in comparison with the group of nr-AxSpA patients and HC, in particular miR-24, miR-27a, miR-106a or miR-223 (p < 0.0001). Levels of these miRNAs were significantly lower (p < 0.05) in a subgroup of patients with bamboo spine compared with other patients with AxSpA. We found, that e.g. miR-625* and miR-885–5p significantly correlated with BASDAI in all patients with r-AxSpA (r = 0.330; p = 0.043 and r = 0.433; p = 0.007 respectively) and were more pronounced in patients with spinal involvement (r = 0.570; p = 0.011 and r = 0.587; p = 0.008). Furthermore, miR-29a significantly correlated with BASDAI (r = 0.474; p = 0.040) and CRP (r = 0.625; p = 0.004) in patients with spinal involvement. Conclusions Our analysis revealed different expression of several circulating miRNAs associated with the degree of spinal involvement and disease activity. Our data suggest role of these miRNA in the pathogenesis of AxSpA and potential use of circulating miRNAs as biomarkers of disease progression. Acknowledgements IGA project no. NT 14498, project of MHCR for conceptual development of research organisation 023728 and SVV 260 031.

A8.06 Microrna-125b expression in PBMCS is inversely associated with disease activity in patients with early rheumatoid arthritis

Background and objectives MicroRNAs (miRNAs) are small RNAs that regulate gene expression by targeting mRNA. It was proved that some miRNAs are significantly deregulated in rheumatoid arthritis (RA). MicroRNA-125b negatively regulates expression of TNF-α, which plays a crucial role in RA pathogenesis. The aim of this study was to evaluate the expression of miRNA-125b in peripheral blood mononuclear cells (PBMCs) and its association with disease activity and treatment response in early RA patients. Materials and methods A total of 58 (42 females; mean age 54.3 ± 16.4 years) early RA patients and 54 (41 females; mean age 50.9 ± 15.11 years) healthy controls (HC) were studied. Total RNA was isolated from PBMCs collected from patients before and after three months of glucocorticoid/disease modifying antirheumatic drugs treatment and from HC. The expression of miRNA-125b was determined by quantitative PCR. RNU44 was used for normalisation. Disease activity was defined using 28-Joint Count Disease activity Score (DAS28-ESR). The expression of miRNA-125b was studied in order to predict treatment response characterised by achieving remission or alternatively low disease activity (DAS28 < 3.2). Results The expression of miRNA-125b was significantly lower in early RA patients compared to HC (p = 0.001) and increased after three months of therapy (p = 0.006), mainly in responders (p = 0.019). At baseline, miRNA-125b expression negatively correlated with DAS28 (r = -0.462; p = 0.0003). In addition, baseline miRNA-125b expression predicted treatment response after three months which was performed by ROC curve analysis (AUC: 0.663 [95% CI 0.520 to 0.805]; p = 0.048). Conclusion Monitoring of miRNA-125b could be used as a biomarker of disease activity and treatment response in early RA. Acknowledgement IGA project No NT 14498; GAUK-367615.

A3.10 Serum calprotectinis elevated in patients with early rheumatoid arthritis but not in patients at risk of developing rheumatoid arthritis

Background and objectives Calprotectin (S100A8/S100A9 complex) is associated with disease activity, radiographic progression and may predict treatment response in patients with rheumatoid arthritis (RA). The aim of our study was to examine the serum levels of calprotectin in clinically suspect arthralgia patientswith positive antibodies to citrullinated peptide antigens (ACPA) who areat high risk of developing RA. Materials and methods This cross-sectional study included 20 ACPA+ arthralgiapatients, 55 patients with early RA (disease duration <6 months who fulfilled 2010 ACR/EULAR classification criteria) and 68 healthy controls (HC). Disease activity was assessed using DAS28. Ultrasound of 28 small joints was performed in all patients with arthralgia to evaluate subclinical synovitis. Serum calprotectin levels were determined byELISA. Data were analysed using 1way ANOVA, Mann-Whitney test and Spearman’s and Pearson’s correlation coefficients. The data are expressed as mean±SD. Results Patients with clinically suspect arthralgia showed no clinical and ultrasound evidence of arthritis (grey scale and power Doppler for single joint ≤ 1). Of these, all were ACPA+, 70%females; age 41.92 ± 11.72 years, CRP 7.39 ± 19.19 mg/l. Treatment naïve patients with early RA had active disease (meanDAS28:5.54 ± 1.62), 65% were ACPA+, 71% were females; age 54.17 ± 16.77 years and CRP 20.46 ± 26.66 mg/l. Serum calprotectin in clinically suspect ACPA+ arthralgia patientswas not significantly different compared to HC (2980 ± 1610 vs. 3368 ± 1624 ng/ml, p = 0.35) but was significantly lower in both groups compared to early RA (5977 ± 5787 ng/ml, p = 0.01 and 0.02 respectively). There was no correlation between calprotectin and CRP in suspect arthralgia patients, however, in early RA, serum calprotectin significantly correlated with DAS28 (r = 0.432, p = 0.001) and CRP (r = 0.670, p < 0.0001). Conclusions Calprotectin is not increased in ACPA positive patients who have normal ultrasound findings of small joints and thus may not help to predict RA development in this early stage of the disease. However, calprotectin is significantly elevated in patients with active early RA, where it serves as a reliable biomarker of disease activity. Acknowledgements IGA project no. NT 14498, project of MHCR 023728 and project SVV 260 155.