Recent progresses in the understanding of facioscapulohumeral muscular dystrophy (FSHD) genetics opened the way to the development of targeted therapies. However, knowledge about pathophysiology of muscle damage is still limited and there is increasing need to identify biomarkers of disease activity in the perspective of clinical trial readiness.
We analyzed inflammatory mediators in the interstitial fluid of muscles with different MRI signal in FSHD patients, comparing muscles displaying early lesions on short-tau inversion recovery (STIR) sequences with normal ones. Patients with one T1-weighted normal and STIR hyperintense (STIR+) and contralateral T1-weighted and STIR normal (STIR-) lower limb muscle were asked to enter the study. Twelve consecutive patients, five controls, and one non-penetrant gene carrier underwent prolonged muscle microdialysis with high cut-off membranes. Microdialysates were analyzed using xMAP technology with a wide panel for cytokines, chemokines, and growth factors. A small number of inflammatory mediators were dysregulated in STIR+ versus STIR- and control muscles: CXCL13, upregulated in STIR+ muscles compared with controls (p < 0.01); CXCL5, downregulated in STIR+ compared with STIR- muscles (p < 0.05); and G-CSF, downregulated in STIR+ muscles compared with controls (p < 0.05). CXCL13 was also upregulated in the STIR+ muscles compared with the contralateral STIR- muscles of the same patient (p < 0.01).
These results support the evidence of a selective inflammatory process taking place in STIR+ FSHD muscles. The application of microdialysis could provide insights on novel mechanisms involved in muscle damage in FSHD and in other myopathies. Further studies are needed to validate these investigated molecules as tissue and circulating biomarkers.
This is a preview of subscription content, log in to check access.
The authors gratefully acknowledge the FSHD Italia ONLUS Association.
Compliance with Ethical Standards
This protocol is in agreement with the Declaration of Helsinki and was approved by the Ethics Committee of our Institution. All involved subjects gave their written informed consent.
This study was supported by a grant from the FSH Society (FSHS-82013-05) and fundings from the Don Carlo Gnocchi ONLUS Foundation, Ricerca corrente 2014, to GT.
Conflict of Interest
Pursuant to the terms of a Master Academic Services Agreement with the Catholic University of the Sacred Heart, M. Monforte and E. Ricci have provided central reading services for MRI scans generated in aTyr’s clinical trials of Resolaris (ATYR1940). E. Ricci is the site principal investigator for some of such trials. The other authors report no disclosures.
Tasca G, Monforte M, Ottaviani P, Pelliccioni M, Frusciante R, Laschena F et al (2016) Magnetic resonance imaging in a large cohort of facioscapulohumeral muscular dystrophy patients: pattern refinement and implications for clinical trials. Ann Neurol. doi:10.1002/ana.24640PubMedGoogle Scholar
Tasca G, Monforte M, Iannaccone E, Laschena F, Ottaviani P, Leoncini E et al (2014) Upper girdle imaging in facioscapulohumeral muscular dystrophy. PLoS One 9:e100292CrossRefPubMedPubMedCentralGoogle Scholar
Kan HE, Scheenen TW, Wohlgemuth M, Klomp DW, van Loosbroek-Wagenmans I, Padberg GW et al (2009) Quantitative MR imaging of individual muscle involvement in facioscapulohumeral muscular dystrophy. Neuromuscul Disord 19:357–362CrossRefPubMedGoogle Scholar
Friedman SD, Poliachik SL, Carter GT, Budech CB, Bird TD, Shaw DW (2012) The magnetic resonance imaging spectrum of facioscapulohumeral muscular dystrophy. Muscle Nerve 45:500–506CrossRefPubMedGoogle Scholar
Frisullo G, Frusciante R, Nociti V, Tasca G, Renna R, Iorio R et al (2011) CD8(+) T cells in facioscapulohumeral muscular dystrophy patients with inflammatory features at muscle MRI. J Clin Immunol 31:155–166CrossRefPubMedGoogle Scholar
Hauerslev S, Ørngreen MC, Hertz JM, Vissing J, Krag TO (2013) Muscle regeneration and inflammation in patients with facioscapulohumeral muscular dystrophy. Acta Neurol Scand 128:194–201CrossRefPubMedGoogle Scholar
Tasca G, Pescatori M, Monforte M, Mirabella M, Iannaccone E, Frusciante R et al (2012) Different molecular signatures in magnetic resonance imaging-staged facioscapulohumeral muscular dystrophy muscles. PLoS One 7:e38779CrossRefPubMedPubMedCentralGoogle Scholar
Rooyackers O, Thorell A, Nygren J, Ljungqvist O (2004) Microdialysis methods for measuring human metabolism. Curr Opin Clin Nutr Metab Care 7:515–521CrossRefPubMedGoogle Scholar
Winter CD, Pringle AK, Clough GF, Church MK (2004) Raised parenchymal interleukin-6 levels correlate with improved outcome after traumatic brain injury. Brain 127:315–320CrossRefPubMedGoogle Scholar
Tawil R, Padberg GW, Shaw DW, van der Maarel SM, Tapscott SJ, FSHD WP (2016) Clinical trial preparedness in facioscapulohumeral muscular dystrophy: clinical, tissue, and imaging outcome measures 29-30 May 2015, Rochester, New York. Neuromuscul Disord 26:181–186CrossRefPubMedGoogle Scholar
Khan IH, Krishnan VV, Ziman M, Janatpour K, Wun T, Luciw PA et al (2009) A comparison of multiplex suspension array large-panel kits for profiling cytokines and chemokines in rheumatoid arthritis patients. Cytometry B Clin Cytom 76:159–168CrossRefPubMedPubMedCentralGoogle Scholar
Gabriëls J, Beckers MC, Ding H, De Vriese A, Plaisance S, van der Maarel SM et al (1999) Nucleotide sequence of the partially deleted D4Z4 locus in a patient with FSHD identifies a putative gene within each 3.3 kb element. Gene 236:25–32CrossRefPubMedGoogle Scholar
Dixit M, Ansseau E, Tassin A, Winokur S, Shi R, Qian H et al (2007) DUX4, a candidate gene of facioscapulohumeral muscular dystrophy, encodes a transcriptional activator of PITX1. Proc Natl Acad Sci U S A 104:18157–18162CrossRefPubMedPubMedCentralGoogle Scholar
Lemmers RJ, van der Vliet PJ, Klooster R, Sacconi S, Camano P, Dauwerse JG et al (2010) A unifying genetic model for facioscapulohumeral muscular dystrophy. Science 329:1650–1653CrossRefPubMedPubMedCentralGoogle Scholar
Snider L, Geng LN, Lemmers RJ, Kyba M, Ware CB, Nelson AM et al (2010) Facioscapulohumeral dystrophy: incomplete suppression of a retrotransposed gene. PLoS Genet 6:e1001181CrossRefPubMedPubMedCentralGoogle Scholar
Geng LN, Yao Z, Snider L, Fong AP, Cech JN, Young JM et al (2012) DUX4 activates germline genes, retroelements, and immune mediators: implications for facioscapulohumeral dystrophy. Dev Cell 22:38–51CrossRefPubMedGoogle Scholar
Figarella-Branger D, Pellissier JF, Serratrice G, Pouget J, Bianco N (1989) Immunocytochemical study of the inflammatory forms of facioscapulohumeral myopathies and correlation with other types of myositis. Ann Pathol 9:100–108PubMedGoogle Scholar
Arahata K, Ishihara T, Fukunaga H, Orimo S, Lee JH, Goto K et al (1995) Inflammatory response in facioscapulohumeral muscular dystrophy (FSHD): immunocytochemical and genetic analyses. Muscle Nerve 2:S56–S66CrossRefPubMedGoogle Scholar
Honda H, Mano Y, Takahashi A (1987) Inflammatory changes in affected muscles of facioscapulohumeral dystrophy. J Neurol 234:408–411CrossRefPubMedGoogle Scholar
Macaione V, Aguennouz M, Rodolico C, Mazzeo A, Patti A, Cannistraci E et al (2007) RAGE-NF-kappaB pathway activation in response to oxidative stress in facioscapulohumeral muscular dystrophy. Acta Neurol Scand 115:115–121CrossRefPubMedGoogle Scholar
Olausson P, Gerdle B, Ghafouri N, Larsson B, Ghafouri B (2012) Identification of proteins from interstitium of trapezius muscle in women with chronic myalgia using microdialysis in combination with proteomics. PLoS One 7:e52560CrossRefPubMedPubMedCentralGoogle Scholar
Li Z-W, Zhao L, Han Q-C, Zhu X (2016) CXCL13 inhibits microRNA-23a through PI3K/AKT signaling pathway in adipose tissue derived-mesenchymal stem cells. Biomed Pharmacother 83:876–880CrossRefPubMedGoogle Scholar
Dmitriev P, Kiseleva E, Kharchenko O, Ivashkin E, Pichugin A, Dessen P et al (2016) Dux4 controls migration of mesenchymal stem cells through the Cxcr4-Sdf1 axis. Oncotarget 7:65090–65108CrossRefPubMedPubMedCentralGoogle Scholar
Legler DF, Loetscher M, Roos RS, Clark-Lewis I, Baggiolini M, Moser B (1998) B cell-attracting chemokine 1, a human CXC chemokine expressed in lymphoid tissues, selectively attracts B lymphocytes via BLR1/CXCR5. J Exp Med 187:655–660CrossRefPubMedPubMedCentralGoogle Scholar
De Paepe B, Creus KK, De Bleecker JL (2009) Role of cytokines and chemokines in idiopathic inflammatory myopathies. Curr Opin Rheumatol 21:610–616CrossRefPubMedGoogle Scholar
Pícha D, Moravcová L, Smíšková D (2016) Prospective study on the chemokine CXCL13 in neuroborreliosis and other aseptic neuroinfections. J Neurol Sci 368:214–220CrossRefPubMedGoogle Scholar
Schiffer L, Worthmann K, Haller H, Schiffer M (2015) CXCL13 as a new biomarker of systemic lupus erythematosus and lupus nephritis - from bench to bedside. Clin Exp Immunol 179:85–89CrossRefPubMedGoogle Scholar
Jones JD, Hamilton BJ, Challener GJ, de Brum-Fernandes AJ, Cossette P, Liang P et al (2014) Serum C-X-C motif chemokine 13 is elevated in early and established rheumatoid arthritis and correlates with rheumatoid factor levels. Arthritis Res Ther 16:R103CrossRefPubMedPubMedCentralGoogle Scholar
Nishikawa A, Suzuki K, Kassai Y, Gotou Y, Takiguchi M, Miyazaki T et al (2016) Identification of definitive serum biomarkers associated with disease activity in primary Sjögren’s syndrome. Arthritis Res Ther 18:106CrossRefPubMedPubMedCentralGoogle Scholar
Zhang Q, Cao DL, Zhang ZJ, Jiang BC, Gao YJ (2016) Chemokine CXCL13 mediates orofacial neuropathic pain via CXCR5/ERK pathway in the trigeminal ganglion of mice. J Neuroinflammation 13:183CrossRefPubMedPubMedCentralGoogle Scholar
Müller G, Lipp M (2003) Concerted action of the chemokine and lymphotoxin system in secondary lymphoid-organ development. Curr Opin Immunol 15:217–224CrossRefPubMedGoogle Scholar
Yang T, Wang S, Zheng Q, Wang L, Li Q, Wei M et al (2016) Increased plasma levels of epithelial neutrophil-activating peptide 78/CXCL5 during the remission of neuromyelitis optica. BMC Neurol 16:96CrossRefPubMedPubMedCentralGoogle Scholar
Zhao Y, Zhang H (2016) Update on the mechanisms of homing of adipose tissue-derived stem cells. Cytotherapy 18:816–827CrossRefPubMedGoogle Scholar
Rando A, Gasco S, de la Torre M, García-Redondo A, Zaragoza P, Toivonen JM et al (2017) Granulocyte colony-stimulating factor ameliorates skeletal muscle dysfunction in amyotrophic lateral sclerosis mice and improves proliferation of SOD1-G93A myoblasts in vitro. Neurodegener Dis 17:1–13CrossRefPubMedGoogle Scholar
Hayashiji N, Yuasa S, Miyagoe-Suzuki Y, Hara M, Ito N, Hashimoto H et al (2015) G-CSF supports long-term muscle regeneration in mouse models of muscular dystrophy. Nat Commun 6:6745CrossRefPubMedGoogle Scholar
Statland J, Donlin-Smith CM, Tapscott SJ, van der Maarel S, Tawil R (2014) Multiplex screen of serum biomarkers in facioscapulohumeral muscular dystrophy. J Neuromuscul Dis 1:181–190PubMedCentralGoogle Scholar