Interleukin-35 (IL-35) is a newly described heterodimeric cytokine that belongs to the IL-12 family and consists of p35 (IL-12a) and EBI3 (IL-27b) subunits. IL-35 exerts immunomodulatory activities in several autoimmune inflammatory diseases.
Objectives
The aim of this study was to assess IL-35 expression in muscle tissue of patients with idiopathic inflammatory myopathies (IIM) and to compare serum levels of IL-35 in patients with IIM to healthy controls and asses potential association with activity of IIM.
Methods
The expression of IL-35 was studied in a series of 19 muscle biopsy samples of idiopathic inflammatory myopathies (9 dermatomyositis, 10 polymyositis) and 10 non-inflammatory control muscle biopsies from patients with myasthenia gravis.Serum levels of IL-35 were measured in 23 PM, 28 DM and 15 cancer associated myositis patients as well as in 40 healthy controls.Disease activity was evaluated by the Myositis Disease Activity Assessment Tool (MYOACT) and by serum muscle enzymes.
Results
Both IL-35 subunits were found in immune cells of the inflammatory infiltrates in IIM muscle biopsies, no immunoreactivity was observed in muscle tissue of control patients.IL-35 serum levels were increased in all IIM patients compared to healthy controls (p<0.001). There were no differences in IL-35 serum levels between myositis subgroups. Serum IL-35 levels correlated with the overall MYOACT score, with extramuscular and muscle domains of MYOACT, with physician9s global activity assessment and lactate dehydrogenase levels.
Conclusions
IL-35 subunits are overexpressed in inflammatory infiltrates in muscle tissue of IIM patients and elevated circulating IL-35 levels correlate with several disease activity parameters. These data suggest potential role of IL-35 in the pathogenesis of inflammatory myopathies.
Acknowledgement
Supported by MHCR support for conceptual development of a research organization (023728) and BTCure (115142–2).
Interleukin-17A inhibitors (IL-17Ai) and tumour necrosis factor inhibitors (TNFi) are available for treatment of axial spondyloarthritis (axSpA). Bio-naïve patients generally respond better to the first TNFi compared to those who are TNFi-experienced. However, real-world data on the effectiveness of IL-17Ai used as a first line treatment vs a second or third line treatment, i.e. after TNFi exposure, are sparse.
Objectives:
In axSpA patients initiating a first IL-17Ai treatment in routine care and stratified by prior exposure to TNFi (0/1/2), we aimed to a) describe the baseline demographics, clinical and disease activity characteristics, b) assess and compare 6-month composite scores of low disease activity (LDA) and response rates, and c) assess and compare 12-month retention rates.
Methods:
The study was conducted within the European Spondyloarthritis (EuroSpA) Research Collaboration Network. Patients with axSpA who initiated a first IL-17Ai between 2018 and 2022 were included from 13 European registries. Logistic regression analyses were employed to compare 6-month LDA (Ankylosing Spondylitis Disease Activity Score using C-reactive protein (ASDAS-CRP) <2.1 and Bath Ankylosing Spondylitis Disease Activity Index (BASDAI) <4) and response rates (ASAS 40 response and ASDAS-CRP Clinically Important Improvement (CII), i.e delta-ASDAS-CRP ≥1.1). Log-rank tests and cox regression analyses were performed to compare 12-month IL-17Ai retention. All analyses were adjusted for age, sex, registry, time since diagnosis, and ASDAS-CRP at treatment start (baseline). To account for missing baseline data, multivariate imputation by chained equations were employed.
Results:
A total of 2265 axSpA patients were included, with 799/787/679 having previously received 0/1/2 TNFi, respectively. At baseline, bio-naïve patients were more likely to be male (64%/ 53%/49% for 0/1/2 previously TNFi-exposed) and HLA-B27 positive (81%/78%/74%). Additionally, bio-naïve patients had higher disease activity according to BASDAI (6.4/5.2/5.8) and ASDAS-CRP (4.2/3.3/3.5), and a higher proportion of them received concomitant csDMARDs (29%/21%/19%). At 6 months, bio-naïve patients demonstrated numerically higher rates of LDA and response across all outcomes compared to TNFi-experienced patients. Similarly, after adjusting for confounders, significantly better responses were found for bio-naïve compared to TNFi-experienced patients for all outcomes (Table 1). The overall 12-month IL-17Ai retention rate was 79.5%. Notably, the 12-month IL-17Ai retention decreased with an increased number of previous TNFi (85.2%/77.9%/74.6%, for 0/1/2 previously TNFi-exposed, p<0.001), with statistically significant differences between bio-naïve and TNFi-experienced patients (Table 1).
Conclusion:
This large real-world study showed overall high one-year retention of IL-17Ai treatment in axSpA. Notably, bio-naïve patients, despite exhibiting higher baseline disease activity, demonstrated superior 6-month LDA and response rates as well as higher 12-month retention compared to TNFi-experienced patients.
Acknowledgements:
The EuroSpA collaboration is supported by Novartis and UCB. This EuroSpA study was financially supported by UCB. UCB had no influence on the data collection, statistical analyses, abstract preparation, or decision to submit.
Disclosure of Interests:
Marion Pons Novartis (paid to the employer), Stylianos Georgiadis Novartis (paid to the employer), Merete Lund Hetland Pfizer, Medac, Sandoz (no personal income, institution), Advisory Board Abbvie (No personal income, paid to institution). Prev. chaired the steering committee of the Danish Rheumatology Quality Registry (DANBIO, DRQ), which receives public funding from the hospital owners and funding from pharmaceutical companies., Research grants (institution) from Abbvie, Biogen, BMS, Celltrion, Eli Lilly, Janssen Biologics B.V, Lundbeck Fonden, MSD, Medac, Pfizer, Roche, Samsung Biopies, Sandoz, Novartis, Nordforsk, Louise Linde Research grant from Novartis, Mehrdad Shoae Kazemi Novartis (paid to the employer), Simon Horskjær Rasmussen Novartis (paid to the employer), Anne Gitte Loft Speaking fees from AbbVie, Janssen, Lilly, MSD, Novartis, Pfizer, UCB, Consulting fees from AbbVie, Janssen, Lilly, MSD, Novartis, Pfizer, UCB, Research grant from Novartis, Brigitte Michelsen Consulting fees from Novartis, Research grant from Novartis (paid to employer). Centre for treatment of Rheumatic and Musculoskeletal Diseases (REMEDY) is funded as a Centre for Clinical Treatment Research by The Research Council of Norway (project 328657), Daniela Di Giuseppe: None declared, Gary J. Macfarlane Research grant from GSK, Gareth T. Jones Speaker fee from Janssen., Research grants (paid to employer) from AbbVie, Pfizer, UCB, Amgen, GSK., Karin Laas Abbvie, Johnson and Johnson, Novartis, Pfizer, Sigrid Vorobjov: None declared, Isabel Castrejon Speaker fees from BMS, Eli-Lilly, Galapagos, Gilead, Janssen, Novartis, MSD, Pfizer, GSK, Consultancy fees from BMS, Eli-Lilly, Galapagos, Gilead, Janssen, Novartis, MSD, Pfizer, GSK, Fernando Sánchez-Alonso: None declared, Ziga Rotar Abbvie, Amgen, Novartis, MSD, Medis, Biogen, Eli Lilly, Pfizer, Sanofi, Lek, Janssen, Abbvie, Novartis, Eli Lilly, Pfizer, Janssen, SOBI, Swixx BioPharma, AstraZeneca, Katja Perdan Pirkmajer Abbvie, Novartis, MSD, Medis, Eli Lilly, Pfizer, Lek, Janssen, Abbvie, Novartis, Medis, Eli Lilly, Pfize, Boehringer Ingelheim, Jana Baranová: None declared, Bente Glintborg Research grants from Pfizer, Abbvie, BMS, Sandoz, Adrian Ciurea: None declared, Miguel Bernardes: None declared, Paula Valente: None declared, Bjorn Gudbjornsson Speaking fees from Novartis and Nordic-Pharma, Consulting fees from Novartis, Gerdur Grondal: None declared, Catalin Codreanu Speaker fees from AbbVie, Amgen, Boehringer Ingelheim, Ewopharma, Lilly, Novartis, Pfizer, Consulting fees from AbbVie, Amgen, Boehringer Ingelheim, Ewopharma, Lilly, Novartis, Pfizer, Corina Mogosan Speaker fees from AbbVie, Boehringer Ingelheim, Ewopharma, Lilly, Novartis, Pfizer, Sella Aarrestad Provan Boehringer Ingelheim, Boehringer Ingelheim, Florenzo Iannone Speaking fees from Abbvie, Amgen, AstraZeneca, BMS, Galapagos, Janssen, Lilly, MSD, Novartis, Pfizer, Roche, UCB, Consulting fees from Abbvie, Amgen, AstraZeneca, BMS, Galapagos, Janssen, Lilly, MSD, Novartis, Pfizer, Roche, UCB, Research grant from BMS, Galapagos, Pfizer, Roberto F. Caporali Speaking fees from Abbvie, Amgen, AstraZeneca, BMS, Galapagos, Janssen, Lilly, MSD, Novartis, Pfizer, Roche, UCB, Consulting fees from Abbvie, Amgen, AstraZeneca, BMS, Galapagos, Janssen, Lilly, MSD, Novartis, Pfizer, Roche, UCB, Johan K Wallman Speaker fees from AbbVie, Amgen, Research support from AbbVie, Amgen, Eli Lilly, Novartis, Pfizer, Vappu Rantalaiho Novartis, Viatris, Pfizer, Ritva Peltomaa Abbvie, Boehringer,Celltrion,Fresenius, Lilly, UCB, Gunnstein Bakland: None declared, Ladislav Šenolt: None declared, Mikkel Østergaard Speaker and/or consultancy fees from Abbvie, BMS, Boehringer-Ingelheim, Celgene, Eli-Lilly, Hospira, Janssen, Merck, Novartis, Novo, Orion, Pfizer, Regeneron, Roche, Sandoz, Sanofi, UCB, Speaker and/or consultancy fees from Abbvie, BMS, Boehringer-Ingelheim, Celgene, Eli-Lilly, Hospira, Janssen, Merck, Novartis, Novo, Orion, Pfizer, Regeneron, Roche, Sandoz, Sanofi, UCB, Research grants from Abbvie, BMS, Merck, Novartis and UCB, Lykke Midtbøll Ørnbjerg Research grant from Novartis.
The autoantibodies such as autoantibodies against citrullinated proteins (ACPA) can be detected years before the first onset of rheumatoid arthritis (RA) and their presence in at-risk individuals conferred increased absolute risks of developing clinical arthritis. Clinical characteristics of patients with arthralgia who are considered at risk for RA (clinically suspect arthralgia, CSA) were defined by EULAR to differentiate these from patients with other types of arthralgia. We have previously described a shift from classical to pro-inflammatory non-classical monocyte subpopulations in the peripheral blood of individuals at risk for RA. CD11c plays a role in the production of pro-inflammatory cytokines and is highly expressed in the mononuclear cells of RA patients.
Objectives:
To study monocyte subpopulations and the expression of CD11c in at-risk individuals with arthralgia.
Methods:
Individuals from the At Risk of RA (ARRA) prospective observational cohort were defined as having arthralgia without arthritis on the examination of 66 joints at baseline and being either ACPA+ and/or meeting the EULAR definition of CSA (further CSA+, fulfilling at least 3 out of 7 parameters). Peripheral blood samples at baseline were analyzed by flow cytometry. Monocytes were classified into classical (CD14++CD16-), intermediate (CD14++CD16+/++), and non-classical (CD14-/+CD16++) subpopulations. The membrane expression of CD11c was assessed in each of these subpopulations as a median of fluorescence. Ultrasound (US) synovitis was evaluated using GLOESS (global OMERACT-EULAR score system) and US7 (the German ultrasonography 7) scores. Data were analyzed using Mann Whitney test and Spearman's correlation coefficient and expressed as median and interquartile range [IQR].
Results:
Out of 207 at-risk individuals with a median symptom duration of 14 months, 46 developed clinical arthritis (progressors) within a median of 8 months of follow-up and after 30.45 [14.55-67.88] months of symptom duration, with CRP 5.50 [2.09-14.10] mg/l and DAS28-CRP score 4.46 [3.10-5.23] at the time of manifestation. Those individuals (n=151) who had not yet progressed to arthritis with symptom duration longer than 12 months at the most recent clinical assessment were defined as non-progressors. In the entire ARRA cohort, there were no statistically significant differences in the percentage of monocyte subpopulations between ACPA+ and ACPA- or CSA+ and CSA- individuals. However, ACPA+ individuals had higher CD11c expression in both classical (p=0.024) and nonclassical (p=0.040) monocyte subpopulations compared to ACPA- individuals. There were no differences in the percentage of monocyte subpopulations between progressors and non-progressors, but a subgroup of progressors who were ACPA+ and met the CSA criteria had significantly higher expression of CD11c in classical (p=0.030) and non-classical (p=0.015) monocytes and a trend towards higher expression of CD11c in intermediate (p=0.062) monocytes compared to all non-progressors. In progressors, the baseline percentage of classical monocytes correlated negatively with baseline assessments: the tender joint count, DAS-CRP score (as per the definition of inclusion criteria in the absence of arthritis), and US-detected subclinical synovitis assessed by both GLOESS and US7 score, whereas the percentage of intermediate and non-classical monocytes correlated positively with all these parameters (Table 1). There were no such correlations of monocyte subpopulations with the abovementioned parameters in non-progressors or progressors at the time of arthritis manifestation.
Conclusion:
We demonstrate higher expression of pro-inflammatory marker CD11c in monocytes of ACPA+ individuals with arthralgia, who are considered to be at high risk of developing RA, especially in ACPA+ progressors meeting the definition for CSA. Moreover, the positive correlation of pro-inflammatory monocyte subsets with subclinical activity preceding the onset of clinical arthritis suggests their potential role in the pathogenesis of RA.
Objective Osteoporosis is associated with an impaired balance between bone resorption and formation, which in turn leads to bone loss and fractures. Many recent studies have underlined the regulatory role of microRNAs (miRNAs) in bone remodeling processes and their potential as biomarkers of osteoporosis. The purpose of this study was to prospectively examine the association of circulating miRNAs and bone biomarkers with estrogen status in women before and after oophorectomy, as well as in oophorectomized women on estrogen therapy. Methods In this prospective study, we included 11 women before oophorectomy and hysterectomy and at 201 ± 24 days after the surgery. Another 11 women were evaluated 508 ± 127 days after oophorectomy and hysterectomy and after an additional 203 ± 71 days of estradiol treatment. Serum miRNAs were profiled by sequencing. Estrogen status and biomarkers of bone metabolism were quantified. Bone mineral density was assessed in the lumbar spine. Results Our analysis revealed 17 miRNAs associated with estrogen levels. Of those miRNAs that were upregulated with estrogen deficiency and downregulated after estrogen therapy, miR-422a correlated with serum beta-carboxy-terminal type I collagen crosslinks (β-CTX) and procollagen 1 N-terminal propeptide (P1NP); and miR-1278 correlated with serum β-CTX, P1NP, osteocalcin, sclerostin, and Dickkopf-1(Dkk1). In contrast, we found an inverse association of miR-24-1-5p with estrogen status and a negative correlation with serum β-CTX, P1NP, osteoprotegerin, and sclerostin levels. Conclusion The reported miRNAs associated with estrogen status and bone metabolism could be potential biomarkers of bone pathophysiology and would facilitate studies on the prevention of postmenopausal osteoporosis. Our findings require validation in an extended cohort.
Systemic sclerosis (SSc) is characterized by fibrosis of the skin and visceral organs, especially digestive tract, and musculoskeletal involvement, which limit mobility/self-sufficiency of patients, and can have a negative impact on body composition.
Objectives
To assess body composition and physical activity of SSc patients and healthy controls (HC).
Methods
59 patients with SSc (50 females, 9 males; mean age 52.1; disease duration 6.7 years; limited cutaneous (lcSSc,36)/diffuse cutaneous (dcSSc,23)) and 36 age-/sex-matched HC (30 females, 6 males, mean age 51.4) without rheumatic/tumor diseases or manifest cardiovascular event were included. SSc patients fulfilled EULAR/ACR 2013 criteria. Anthropometric parameters and body composition were assessed (by densitometry-iDXA Lunar, and by bioelectric impedance-BIA-2000-M), and physical activity was evaluated using Human Activity Profile (HAP) questionnaire. Routine biochemistry analysis was performed after 8 hours of fasting. Disease activity was evaluated by EUSTAR SSc activity score. Data are presented as mean±SD.
Results
Compared to HC, patients with SSc had significantly lower body-mass index (BMI: 26.4±3.3 vs. 22.4±4.3 kg/m2, p<0.0001) and body fat % assessed by both iDXA (BF%: 37.2±6.6 vs. 32.6±8.2%, p=0.0014) and BIA (BF%: 31.1±6.4 vs. 24.6±7.8%, p<0.0001), and a trend to decreased visceral fat weight (0.9±0.9 vs. 0.5±0.5kg, p=0.0670). Compared to HC, SSc patients demonstrated significantly decreased lean body mass assessed by both iDXA (LBM: 46.6±7.5 vs. 40.9±6.8kg, p=0.0003) and BIA (LBM: 53.2±8.7 vs. 47.7±7.0kg, p=0.0017), and increased ECM/BCM ratio (extracellular mass/body cell mass: 1.03±0.1 vs. 1.29±0.4, p<0.0001), which reflects worse muscle predispositions for physical exercise, aerobic fitness/performance, and usually increases with deteriorating nutritional status. Compared to HC, SSc patients had significantly lower bone mineral density (BMD: 1.16±0.10 vs. 1.05±0.11g/cm2, p<0.0001), and were currently able to perform less energetically demanding physical activities according to HAP score (84.7±6.6 vs. 64.1±17.2, p<0.0001). Disease activity negatively correlated with BF% (r=-0.324, p=0.014), and physical activity (HAP) positively correlated with BMD (r=0.276, p=0.034) and negatively with ECM/BCM (r=-0.625, p<0.0001).
Conclusions
Compared to healthy age-/sex-matched individuals we found significant negative changes in body composition of our SSc patients, which are associated with their disease activity and physical activity, and could reflect their nutritional status, and gastrointestinal and musculoskeletal involvement.
Background: Heat shock proteins (Hsps) are chaperones playing important roles in skeletal muscle physiology, adaptation to exercise or stress, and activation of inflammatory cells Objectives: The aim of our study was to assess Hsp90 expression in muscle biopsies and plasma of patients with idiopathic inflammatory myopathies (IIM) and to characterize its association with IIM-related features. Methods: Total of 277 patients with IIM (198 females, 79 males; mean age 54.8; disease duration 4.1 years; DM, 104/PM, 108/CADM, 31/IMNM, 25) and 157 healthy individuals (92 females, 65 males; mean age 47.0) were included in plasma analysis. Muscle biopsy samples (PM, DM, IMNM, myodystrophy, myasthenia gravis) were stained for Hsp90α (Thermo Fisher Scientific, USA) and Hsp90β (Abcam, UK). Plasma Hsp90 was measured by ELISA kit (eBioscience, Vienna, Austria). The cytokines/chemokines were analysed by using Bio-Plex Pro TM human Cytokine 27-plex Assay (BIO-RAD, California, USA.Data are presented as median(IQR). Results: In muscle biopsies, Hsp90 expression of both subunits (alpha and beta) was higher in IIM than in controls. Increased Hsp90 was detected in perifascicular degenerating and regenerating fibers, inflammatory cells (DM, PM), and necrotic and regenerating fibers (IMNM). Plasma Hsp90 levels were increased in IIM patients compared to healthy controls (55.9 (46.9 – 62.5)vs 9.76(7.5 – 13.8), p<0.0001), and in individual subgroups of IIM vs. healthy controls (DM-22.01(14.1 – 41.2), PM-19.7(14.3 – 42.2), CADM-18.9(11.7 – 29.7), IMNM-19.6(16.3 – 45.5), p<0.0001 for all). Hsp90 was higher in males compared to females (p=0.040) and in patients with ILD (p=0.003), cardiac involvement (p=0.004), dysphagia (p=0.018) and presence of anti-Ro52 (p=0.036). Hsp90 levels in all patients positively correlated with muscle enzymes (Tab.1). Hsp90 was associated with disease activity and skeletal muscle involvement (Tab.1). Out of all clinical parameters listed in above-mentioned univariate analysis, in multiple regression analysis Hsp90 levels in IIM patients were significantly affected by muscle enzymes only (p<0.0001, β=0.345). Furthermore, Hsp90 positively correlated with some crucial cytokines involved in pathogenesis of myositis (Tab. 1). Tab 1 Clinical parameters Spearman’s r p – value LDH; AST; ALT 0.554; 0.383; 0.181 < 0.0001; < 0.0001; 0.003 PtDGA; PhDGA; MITAX; MYOACT 0.223; 0.217; 0.175; 0.159 < 0.001; < 0.001; 0.004; 0.012 Pulmonary disease activity 0.201 0.001 Muscle disease activity 0.146 0.018 MMT8, total score; m. biceps brachii; m. gluteus maximus; m. iliopsoas -0.126; -0.125; -0.159; -0.143 0.042; 0.043; 0.011; 0.023 MDI – Myositis damage index – severity 0.150 0.041 Current Prednisone equivalent dose 0.183 0.006 Cytokines: IL-1b; IL-2; IL-4; IL-6; IFN-γ 0.188; 0.269; 0.190; 0.182; 0.229 0.002; < 0.0001; 0.002; 0.003; < 0.0001 Conclusion: We demonstrate increased Hsp90 expression in IIM muscle biopsy samples, specifically in inflammatory cells, degenerating, regenerating and/or necrotic fibers. Increased Hsp90 plasma levels in IIM patients are associated with disease activity and damage, and with the involvement of proximal skeletal muscles, heart and lungs. Acknowledgments: Supported by AZV-16-33542A, MHCR 023728 and SVV – 260373. Disclosure of Interests: Hana Štorkánová: None declared, Sabina Oreska: None declared, Maja Špiritović: None declared, Barbora Heřmánková: None declared, Olga Kryštůfková: None declared, Heřman Mann: None declared, Martin Komarc: None declared, Josef Zámečník: None declared, Karel Pavelka Consultant of: Abbvie, MSD, BMS, Egis, Roche, UCB, Medac, Pfizer, Biogen, Speakers bureau: Abbvie, MSD, BMS, Egis, Roche, UCB, Medac, Pfizer, Biogen, Jiří Vencovský: None declared, Ladislav Šenolt: None declared, Michal Tomcik: None declared
