Target organ expression and biomarker characterization of chemokine CCL21 in systemic sclerosis associated pulmonary arterial hypertension

Introduction: Systemic sclerosis (SSc) is a heterogenous disorder that appears to result from interplay between vascular pathologies, tissue fibrosis and immune processes, with evidence for deregulation of chemokines, which normally control immune trafficking. We recently identified altered levels of chemokine CCL21 in SSc associated pulmonary arterial hypertension (PAH). Here, we aimed to define target organ expression and biomarker characteristics of CCL21. Materials and methods: To investigate target organ expression of CCL21, we performed immunohistochemistry (IHC) on explanted lung tissues from SSc-PAH patients. We assessed serum levels of CCL21 by ELISA and Luminex in two well-characterized SSc cohorts from Oslo (OUH, n=552) and Zurich (n=93) University hospitals and in 168 healthy controls. For detection of anti-CCl21 antibodies, we performed protein array analysis applying serum samples from SSc patients (n=300) and healthy controls. To characterize circulating CCL21 in SSc, we applied immunoprecipitation (IP) with antibodies detecting both full length and tailless and a custom-made antibody detecting only the C-terminal of CCL21. IP products were analyzed by SDS-PAGE/western blot and Mass spectrometry (MS). Results: By IHC, we found that CCL21 was mainly expressed in the airway epithelial cells of SSc patients with PAH. In the analysis of serum levels of CCL21 we found weak correlation between Luminex and ELISA (r=0.515, p<0.001). Serum levels of anti-CCL21 antibodies were higher in SSc patients than in healthy controls (p<0.001), but only 5% of the SSc population were positive for anti-CCL21 antibodies in SSc, and we found no correlation between anti-CCl21 and serum levels of CCL21. By MS, we only identified peptides located within amino acid (aa) 23-102 of CCL21, indicating that CCL21 in SSc circulate as a truncated protein without the C-terminal tail. Conclusion: This study demonstrates expression of CCL21 in epithelial lung tissue from SSc patients with PAH, and indicate that CCL21 in SSc circulates as a truncated protein. We extend previous observations indicating biomarker potential of CCL21, but find that Luminex is not suitable as platform for biomarker analyses. Finally, in vivo generated anti-CCL21 antibodies exist in SSc, but do not appear to modify serum CCL21 levels in patients with SSc-PAH

Survival and cancer risk in an unselected and complete Norwegian idiopathic inflammatory myopathy cohort

Objective To utilise an exposed/unexposed cohort strategy for mortality and cancer analyses across unselected and complete cohorts of patients with idiopathic inflammatory myopathy (IIM) resident in south-east Norway (denominator population 2.6 million), between 2003 and 2012. Method IIM cases were identified by comprehensive searches through patient administrative databases followed by manual chart review. Polymyositis (PM) and dermatomyositis (DM) cases were classified by the Peter and Bohan and/or Targoff diagnostic criteria and sporadic inclusion body myositis (sIBM) by the European NeuroMuscular Centre (ENMC) criteria from 1997 and/or 2011. Every patient was matched for sex, age and residential area with 15 unexposed/non-IIM individuals drawn from the national population registry. Results Total mortality in the IIM cohort was 27% (87/326). Standardized mortality rate (SMR) was higher in DM (2.6) than PM (2.4) and sIBM (1.7). IIM-related causes of death were frequent (64%) and included cancer (all IIM subsets), aspiration (sIBM), pulmonary complications (PM/DM) and infections (PM/DM). Multivariate analyses identified age at diagnosis (PM and sIBM), positive anti-SSA (PM), cancer (DM), and DLCO < 60% (DM) as independent mortality risk factors. Cancer risk was increased in DM (standard incidence rate 2.0) and PM (SIR = 1.3), but not in sIBM (SIR = 0.9). Ovarian cancer was more prevalent in DM than in the general population (8.3% vs 1.1%). Conclusion Our results suggest that mortality rates and cancer risk remain elevated in DM, and to a lesser degree also in PM. Mortality rate was also increased in sIBM, but some deaths appeared to be due to potentially preventable causes

Generalized bone loss in early rheumatoid arthritis patients followed for ten years in the biologic treatment era

Background: Osteoporosis is a well-known extra articular manifestation in rheumatoid arthritis (RA). Biologic disease modifying anti rheumatic drugs (DMARDs) has been shown to be superior to synthetic DMARDs to reduce bone destruction including generalized bone loss in RA. Our aim was to study short- and long term changes in hip and spine bone mineral density (BMD) in early RA patients treated during the first decade with available biologic DMARDs. Methods: RA patients diagnosed at an out-patient clinic between 1999 and 2001 were consecutively enrolled. Demographic, disease and treatment data were collected and BMD was assessed by dual energy X-ray absorptiometry at baseline and after 2, 5 and 10 years. Results: The 92 included RA patients had a baseline mean age (SD) of 50.9 (13.3) years and symptom duration of 12.4 (6.7) months, 62.0% were women and 66.3% were RF positive. In the first 2 years ever use of biologic DMARDs was 18.5%, synthetic DMARDs 91.3% and prednisolone 62.0% whereas the figures for the subsequent 8 years were 62.6%, 89.2% and 51.4%, respectively. The annual rate of BMD loss in the first 2 years and the subsequent 8 years was at femoral neck −1.00% vs. −0.56%, at total hip −0.96% vs. −0.41% and at spine L1−4 -0.42% vs. 0.00%. Conclusions: Our study adds evidence that aggressive anti-inflammatory treatment including biologic DMARDs reduces the rate of bone loss in RA. Indicating that the burden of osteoporosis is reduced in RA patients treated in clinical practice in the new millennium

Additional file 1: Figure S1. of Comparative analyses of muscle MRI and muscular function in anti-synthetase syndrome patients and matched controls: a cross-sectional study

showing the distribution of myositis subsets in ASS patients with and without anti-Jo1 antibodies. Blue polymyositis (PM), light blue dermatomyositis (DM), red clinical amyopathic dermatomyositis (CADM) and yellow no myositis. (TIF 31 kb

Performance of Candidate Serum Biomarkers for Systemic Sclerosis?Associated Interstitial Lung Disease

Objective Interstitial lung disease (ILD ) in systemic sclerosis (SS c) runs a highly variable course, and prediction tools are highly desired. The aim of this study was to assess the diagnostic and prognostic performance of 4 candidate serum biomarkers for SS c‐associated ILD . Methods Serum samples from a combined cohort of SS c patients (from Paris, France and Oslo, Norway; n = 427) were analyzed by enzyme‐linked immunosorbent assay for concentrations of lung epithelial–derived surfactant protein D (SP ‐D), Krebs von den Lungen 6 glycoprotein (KL ‐6), CCL 18, and OX 40 ligand (OX 40L). Lung fibrosis was measured by high‐resolution computed tomography and pulmonary function tests. Associations of these candidate biomarkers with baseline disease involvement and prediction of disease progression over time (mean ± SD follow‐up 3.2 ± 4.4 years) were investigated. Results In SS c patients at baseline, serum levels of KL ‐6 correlated with the forced vital capacity (FVC ) (r = −0.317, P 10% decrease in the FVC (hazard ratio [HR ] 2.90, 95% CI 1.25–6.73; P = 0.014) and de novo development of extensive disease (HR 3.71, 95% CI 1.02–13.52; P = 0.048). Matrix‐based logistic regression models for the diagnosis and prognosis of SS c‐associated ILD were constructed, and these models discriminated 3 groups of risk (mild, moderate, or high) for the diagnosis or worsening of lung fibrosis according to the serum levels of SP ‐D (for diagnosis) and serum levels of CCL 18 (for progression of disease). Conclusion These results show that SP ‐D is a relevant diagnostic biomarker for SS c‐associated ILD , whereas KL ‐6 could be used to assess the severity of lung fibrosis. CCL 18 appears to be a potential predictive marker for progression of ILD in SS c

Left ventricular diastolic dysfunction predicts mortality in patients with systemic sclerosis

Background Primary cardiac affection is common and is a major cause of death in systemic sclerosis (SSc), but there are knowledge gaps regarding the effect of cardiac dysfunction on mortality. Objectives The purpose of this study was to evaluate diastolic function in a large, unselected SSc cohort and assess the effect of diastolic dysfunction (DD) on mortality. Methods SSc patients followed prospectively at the Oslo University Hospital from 2003 to 2016 with available echocardiographies and matched control subjects were included. DD was assessed by echocardiography according to the 2016 American Society of Echocardiography/European Association of Cardiovascular Imaging guidelines. Pulmonary hypertension (PH) was diagnosed by right heart catheterization. Vital status was available for all patients. Cox regression analyses with hazards ratios (HRs) were conducted. Results Diastolic function was assessed in 275 SSc patients at baseline and in 186 patients at follow-up. At baseline, 46 of the 275 SSc patients (17%) were diagnosed with DD and 195 (71%) had normal diastolic function. After a median follow-up of 3.4 years (interquartile range: 1.6 to 6.2 years), the proportion of DD increased from 17% to 29%. During follow-up, 57% of patients with DD at baseline died, compared with 13% of patients with normal diastolic function. At baseline, 86 patients had performed right heart catheterization, and 43 were diagnosed with PH; of these 60% deceased. In multivariable Cox regression analyses, DD was a stronger predictor of death (HR: 3.7; 95% CI: 1.69 to 8.14; c-index 0.89) than PH (HR: 2.0; 95% CI: 1.1 to 3.9; c-index 0.84). Conclusions DD is frequent in SSc, and the presence of DD is associated with high mortality. DD exceeds PH with respect to predicting mortality

Differential sensitivity of the 2020 revised comprehensive diagnostic criteria and the 2019 ACR/EULAR classification criteria across IgG4-related disease phenotypes: results from a Norwegian cohort

Abstract Background We investigated sensitivity of the 2020 Revised Comprehensive Diagnostic Criteria (RCD) and the 2019 ACR/EULAR classification criteria across the four identified IgG4-related disease (IgG4-RD) phenotypes: “Pancreato-Hepato-Biliary”, “Retroperitoneum and Aorta”, “Head and Neck-limited” and “Mikulicz’ and Systemic” in a well-characterized patient cohort. Methods We included adult patients diagnosed with IgG4-RD after comprehensive clinical assessment at Oslo University Hospital in Norway. We assigned patients to IgG4-RD phenotypes based on pattern of organ involvement and assessed fulfillment of RCD and 2019 ACR/EULAR classification criteria. Differences between phenotype groups were analyzed using one-way ANOVA for continuous variables, and contingency tables for categorical variables. Results The study cohort included 79 IgG4-RD patients assigned to the “Pancreato-Hepato-Biliary” (22.8%), Retroperitoneum and Aorta” (22.8%) “Head and Neck-limited” (29.1%), and “Mikulicz’ and Systemic” (25.3%) phenotype groups, respectively. While 72/79 (91.1%) patients in total fulfilled the RCD, proportion differed across phenotype groups and was lowest in the “Retroperitoneum and Aorta” group (66.7%, p < 0.001). Among the 57 (72.2%) patients meeting the 2019 ACR/EULAR classification criteria, proportion was again lowest in the “Retroperitoneum and Aorta” group (27.8%, p < 0.001). Conclusion The results from this study indicate that IgG4-RD patients having the “Retroperitoneum and Aorta” phenotype less often fulfill diagnostic criteria and classification criteria than patients with other IgG4-RD phenotypes. Accordingly, this phenotype is at risk of being systematically selected against in observational studies and randomized clinical trials, with potential implications for patients, caregivers and future definitions of IgG4-RD