Idiopathic inflammatory myopathies – increased expression of heat shock protein-90 in muscle tissue and plasma correlates with disease activity and skeletal muscle involvement

Background:Heat shock proteins (Hsps) are chaperones playing important roles in skeletal muscle physiology, adaptation to exercise or stress, and activation of inflammatory cellsObjectives: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 ProTMhuman 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 1Clinical parametersSpearman’s rp – valueLDH; AST; ALT0.554; 0.383; 0.181< 0.0001; < 0.0001; 0.003PtDGA; PhDGA; MITAX; MYOACT0.223; 0.217; 0.175; 0.159< 0.001; < 0.001; 0.004; 0.012Pulmonary disease activity0.2010.001Muscle disease activity0.1460.018MMT8, total score; m. biceps brachii; m. gluteus maximus; m. iliopsoas-0.126; -0.125; -0.159; -0.1430.042; 0.043; 0.011; 0.023MDI – Myositis damage index – severity0.1500.041Current Prednisone equivalent dose0.1830.006Cytokines:IL-1b; IL-2; IL-4; IL-6; IFN-γ0.188; 0.269; 0.190; 0.182; 0.2290.002; < 0.0001; 0.002; 0.003; < 0.0001Conclusion: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

Download Full-text

OP0138 CLUSTERIN ASSOCIATES WITH DISEASE MECHANISMS AND INFLAMMATION IN MYOSITIS PATIENTS

Annals of the Rheumatic Diseases ◽

10.1136/annrheumdis-2020-eular.6237 ◽

2020 ◽

Vol 79 (Suppl 1) ◽

pp. 89.2-89

Author(s):

T. Kropackova ◽

L. Vernerová ◽

H. Štorkánová ◽

V. Horvathova ◽

M. Vokurková ◽

...

Keyword(s):

Skeletal Muscle ◽

Disease Activity ◽

Muscle Tissue ◽

Tissue Injury ◽

Cytokine Profile ◽

Disease Assessment ◽

Idiopathic Inflammatory Myopathies ◽

Inflammatory Myopathies ◽

Protective Function ◽

Disease Mechanisms

Background:Idiopathic inflammatory myopathies (IIM, myositis) are a heterogeneous group of autoimmune muscle disorders characterized by skeletal muscle weakness and damage, inflammation and extramuscular manifestations. Recent findings suggest that immunological as well as nonimmunological processes, such as endoplasmic reticulum stress, hypoxia, mitochondrial and metabolic dysfunction are involved in the pathogenesis of IIMs [1]. Clusterin (CLU) has been reported to play a protective function in the development of tissue injury, inflammation and autoimmunity, and is involved in the maintenance of immune homeostasis [2].Objectives:This study aimed to explore a potential involvement of the circulating levels and skeletal muscle expression of CLU in pathogenic mechanisms of IIM.Methods:A total of 85 IIM patients and 86 healthy controls (HC) were recruited. In addition, 20 IIM patients and 21 HC underwent a muscle biopsy. Circulating concentrations of CLU were measured by ELISA. Serum cytokine profile of patients and HC was assessed by Cytokine 27-plex Assay. Immunohistochemical localisation of CLU was assessed in 10 IIM and 4 control muscle tissue specimens. The expression of CLU and myositis related cytokines in muscle tissue was determined by real-time PCR.Results:We observed a significant increase of circulating CLU in all IIM patients compared to HC (86.2 (71.6-99.0) vs. 59.6 (52.6-68.4) μg/mL, p < 0.0001). Moreover, CLU serum levels were positively correlated with myositis disease activity assessment (MYOACT) (r = 0.337, p = 0.008), myositis intention-to-treat activity index (MITAX) (r = 0.357, p = 0.004) and global disease assessment evaluated by physician (r = 0.309, p = 0.015). In addition to that, a multivariate redundancy analysis revealed a combined effect of serum CLU and cytokine profile (represented by cytokines and chemokines known to be involved in IIM) on disease activity measures. In muscle tissue, CLU mRNA was significantly increased in IIM patients compared to controls (p = 0.032) and correlated with IL-1β (r = 0.489, p = 0.034), IL-6 (r = 0.581, p = 0.009), TNF (r = 0.485, p = 0.035) and PGC-1α (r = 0.709, p = 0.001) mRNA. Immunohistochemistry revealed CLU accumulation in the cytoplasm of regenerating myofibers.Conclusion:Our results show an up-regulation of clusterin in circulation and skeletal muscle of IIM patients that associates with disease activity and inflammation, and its specific expression in regenerating myofibres. Based on our data and the known cytoprotective function of CLU we suggest an attempt of the organism to limit further muscle damage induced by myositis disease mechanisms.References:[1]Ernste FC, Reed AM. Idiopathic inflammatory myopathies: current trends in pathogenesis, clinical features, and up-to-date treatment recommendations. Mayo Clin Proc. 2013;88:83-105.[2]Savkovic V, Gantzer H, Reiser U, Selig L, Gaiser S, Sack U, et al. Clusterin is protective in pancreatitis through anti-apoptotic and anti-inflammatory properties. Biochem Biophys Res Commun. 2007;356:431-7.Acknowledgments:This work was supported by GAUK 534217 and the Ministry of Health of the Czech Republic grants nr. 16-33746A and 16-33574A.Disclosure of Interests:None declared

Download Full-text

Alterations in activin A–myostatin–follistatin system associate with disease activity in inflammatory myopathies

Rheumatology ◽

10.1093/rheumatology/kez651 ◽

2020 ◽

Vol 59 (9) ◽

pp. 2491-2501 ◽

Cited By ~ 4

Author(s):

Lucia Vernerová ◽

Veronika Horváthová ◽

Tereza Kropáčková ◽

Martina Vokurková ◽

Martin Klein ◽

...

Keyword(s):

Skeletal Muscle ◽

Disease Activity ◽

Muscle Tissue ◽

Activin A ◽

Muscle Disease ◽

Manual Muscle Testing ◽

Healthy Controls ◽

Inflammatory Myopathies ◽

Muscle Testing ◽

Tgf Β1

Abstract Objectives The aim of this study was to investigate the systemic and skeletal muscle levels of atrophy-associated myokines in patients with idiopathic inflammatory myopathies (IIM) and their association with clinical characteristics of myositis. Methods A total of 94 IIM patients and 162 healthy controls were recruited. Of those, 20 IIM patients and 28 healthy controls underwent a muscle biopsy. Circulating concentrations of myostatin, follistatin, activin A and TGF-β1 were assessed by ELISA. The expression of myokines and associated genes involved in the myostatin signalling pathway in muscle tissue was determined by real-time PCR. Results We report decreased levels of circulating myostatin (median 1817 vs 2659 pg/ml; P = 0.003) and increased follistatin (1319 vs 1055 pg/ml; P = 0.028) in IIM compared with healthy controls. Activin A levels were also higher in IIM (414 vs 309 pg/ml; P = 0.0005) compared with controls. Myostatin was negatively correlated to muscle disease activity assessed by physician on visual analogue scale (MDA) (r = −0.289, P = 0.015) and positively to manual muscle testing of eight muscles (r = 0.366, P = 0.002). On the other hand, follistatin correlated positively with MDA (r = 0.235, P = 0.047). Gene expression analysis showed higher follistatin (P = 0.003) and myostatin inhibitor follistatin-like 3 protein (FSTL3) (P = 0.008) and lower expression of activin receptor type 1B (ALK4) (P = 0.034), signal transducer SMAD3 (P = 0.023) and atrophy marker atrogin-1 (P = 0.0009) in IIM muscle tissue compared with controls. Conclusion This study shows lower myostatin and higher follistatin levels in circulation and attenuated expression of myostatin pathway signalling components in skeletal muscle of patients with myositis, a newly emerging pattern of the activin A–myostatin–follistatin system in muscle wasting diseases.

Download Full-text