Plasma C18:0‐ceramide is a novel potential biomarker for disease severity in myasthenia gravis

Myasthenia gravis (MG) is an antibody‐mediated autoimmune disorder characterized by fluctuation of fatigue and weakness of muscle. Due to the heterogeneity of the course of MG, available biomarkers for prognostic prediction are urgently needed. Ceramide (Cer) was reported to participate in immune regulation and many autoimmune diseases, but its effects on MG remain undefined. This study aimed to investigate the ceramides expression levels in MG patients and their potential as novel biomarkers of disease severity. Levels of plasma ceramides were determined by ultra performance liquid chromatography–tandem mass spectrometry (UPLC‐MS/MS). Severity of disease was assessed by quantitative MG scores (QMGs), MG‐specific activities of daily living scale (MG‐ADLs) and 15‐item MG quality of Life (MG‐QOL15). The concentrations of serum interleukin‐1β (IL‐1β), IL‐6, IL‐17A, and IL‐21 were determined by enzyme‐linked immunosorbent assay (ELISA), and the proportions of circulating memory B cells and plasmablasts were detected by flow‐cytometry assay. Four plasma ceramides levels we studied were detected higher in MG patients. And three of them (C16:0‐Cer, C18:0‐Cer, and C24:0‐Cer) were positively associated with QMGs. In addition, receiver operating characteristic (ROC) analysis suggested that plasma ceramides have a good ability of differentiating MG from HCs. Importantly, only C18:0‐Cer was shown to be positively associated with the concentration of serum IL and circulating memory B cells, and the decrease in plasma C18:0‐Cer paralleled the clinical improvement of patients with MG. All together, our data suggest that ceramides may play an important role in the immunopathological mechanism of MG, and C18:0‐Cer has the potential to be a novel biomarker for disease severity in MG.

[1]  H. Stark,et al.  Ceramide synthase 6 impacts T-cell allogeneic response and graft-versus-host disease through regulating N-RAS/ERK Pathway , 2022, Leukemia.

[2]  Dong Guo,et al.  Natural killer cells promote the differentiation of follicular helper T cells instead of inducing apoptosis in myasthenia gravis. , 2021, International immunopharmacology.

[3]  Ó. Rolfsson,et al.  Cerebrospinal Fluid C18 Ceramide Associates with Markers of Alzheimer’s Disease and Inflammation at the Pre- and Early Stages of Dementia , 2021, Journal of Alzheimer's disease : JAD.

[4]  M. Ticchioni,et al.  Memory B Cells Predict Relapse in Rituximab-Treated Myasthenia Gravis , 2021, Neurotherapeutics.

[5]  S. Crotty,et al.  Bcl6-Mediated Transcriptional Regulation of Follicular Helper T cells (TFH). , 2021, Trends in immunology.

[6]  H. Ueno,et al.  Immune Skew of Circulating Follicular Helper T Cells Associates With Myasthenia Gravis Severity , 2021, Neurology: Neuroimmunology & Neuroinflammation.

[7]  Nadeem Shafique Butt,et al.  Exacerbation Rate in Generalized Myasthenia Gravis and Its Predictors , 2020, European Neurology.

[8]  F. Shi,et al.  Incidence, mortality, and economic burden of myasthenia gravis in China: A nationwide population-based study , 2020, The Lancet regional health. Western Pacific.

[9]  H. Wiendl,et al.  Post-intervention Status in Patients With Refractory Myasthenia Gravis Treated With Eculizumab During REGAIN and Its Open-Label Extension , 2020, Neurology.

[10]  Shigeaki Suzuki,et al.  Roles of cytokines and T cells in the pathogenesis of myasthenia gravis , 2020, Clinical and experimental immunology.

[11]  S. Hammad,et al.  Sphingolipids and Diagnosis, Prognosis, and Organ Damage in Systemic Lupus Erythematosus , 2020, Frontiers in Immunology.

[12]  Weibin Liu,et al.  Quantitative features and clinical significance of two subpopulations of AChR-specific CD4+ T cells in patients with myasthenia gravis. , 2020, Clinical immunology.

[13]  F. Piehl,et al.  Comparison Between Rituximab Treatment for New-Onset Generalized Myasthenia Gravis and Refractory Generalized Myasthenia Gravis. , 2020, JAMA neurology.

[14]  Y. Wan,et al.  Molecular control of pathogenic Th17 cells in autoimmune diseases. , 2020, International immunopharmacology.

[15]  J. Pfeilschifter,et al.  Blood ceramides as novel markers for renal impairment in systemic lupus erythematosus. , 2019, Prostaglandins & other lipid mediators.

[16]  R. Mantegazza,et al.  Myasthenia gravis: from autoantibodies to therapy , 2018, Current opinion in neurology.

[17]  N. Ferreirós,et al.  The relevance of ceramides and their synthesizing enzymes for multiple sclerosis. , 2018, Clinical science.

[18]  A. Bai,et al.  Acid sphingomyelinase mediates human CD4+ T-cell signaling: potential roles in T-cell responses and diseases , 2017, Cell Death and Disease.

[19]  Min Hee Kim,et al.  Hepatic inflammatory cytokine production can be regulated by modulating sphingomyelinase and ceramide synthase 6. , 2017, International journal of molecular medicine.

[20]  F. Shi,et al.  Augmentation of Circulating Follicular Helper T Cells and Their Impact on Autoreactive B Cells in Myasthenia Gravis , 2016, The Journal of Immunology.

[21]  P. Münzer,et al.  Acid Sphingomyelinase (ASM) is a Negative Regulator of Regulatory T Cell (Treg) Development , 2016, Cellular Physiology and Biochemistry.

[22]  M. Parnham,et al.  The enigma of ceramide synthase regulation in mammalian cells. , 2016, Progress in lipid research.

[23]  S. Kleinstein,et al.  Autoreactive T Cells from Patients with Myasthenia Gravis Are Characterized by Elevated IL-17, IFN-γ, and GM-CSF and Diminished IL-10 Production , 2016, The Journal of Immunology.

[24]  D. Geerts,et al.  Acid Sphingomyelinase–Derived Ceramide Regulates ICAM-1 Function during T Cell Transmigration across Brain Endothelial Cells , 2016, The Journal of Immunology.

[25]  K. Willecke,et al.  Exacerbation of experimental autoimmune encephalomyelitis in ceramide synthase 6 knockout mice is associated with enhanced activation/migration of neutrophils , 2015, Immunology and cell biology.

[26]  Nils Erik Gilhus,et al.  Myasthenia gravis: subgroup classification and therapeutic strategies , 2015, The Lancet Neurology.

[27]  A. Thiel,et al.  IL‐17‐producing CD4+ T cells contribute to the loss of B‐cell tolerance in experimental autoimmune myasthenia gravis , 2015, European journal of immunology.

[28]  N. Ferreirós,et al.  Lack of ceramide synthase 2 suppresses the development of experimental autoimmune encephalomyelitis by impairing the migratory capacity of neutrophils , 2015, Brain, Behavior, and Immunity.

[29]  Yujiang Fang,et al.  The role of IL-21 in immunity and cancer. , 2015, Cancer letters.

[30]  J. Stiban,et al.  Regulation of ceramide channel formation and disassembly: Insights on the initiation of apoptosis , 2015, Saudi journal of biological sciences.

[31]  G. Wolfe,et al.  Treatment-Refractory Myasthenia Gravis , 2014, Journal of clinical neuromuscular disease.

[32]  J. Sieb.  Myasthenia gravis: an update for the clinician , 2014, Clinical and experimental immunology.

[33]  D. Geng,et al.  RNA interference targeting Bcl-6 ameliorates experimental autoimmune myasthenia gravis in mice , 2014, Molecular and Cellular Neuroscience.

[34]  D. Cho,et al.  C6-ceramide enhances Interleukin-12-mediated T helper type 1 cell responses through a cyclooxygenase-2-dependent pathway. , 2012, Immunobiology.

[35]  A. Rudensky,et al.  Regulatory T cells: mechanisms of differentiation and function. , 2012, Annual review of immunology.

[36]  Hulun Li,et al.  Activation of the receptor for advanced glycation end products (RAGE) exacerbates experimental autoimmune myasthenia gravis symptoms. , 2011, Clinical immunology.

[37]  A. Kimura,et al.  IL‐6: Regulator of Treg/Th17 balance , 2010, European journal of immunology.

[38]  Y. Hannun,et al.  The acid sphingomyelinase/ceramide pathway: biomedical significance and mechanisms of regulation. , 2010, Current molecular medicine.

[39]  E. Gulbins,et al.  The role of sphingolipids and ceramide in pulmonary inflammation in cystic fibrosis. , 2010, The open respiratory medicine journal.

[40]  G. Nixon.  Sphingolipids in inflammation: pathological implications and potential therapeutic targets , 2009, British journal of pharmacology.

[41]  G. Samsa,et al.  Quantitative myasthenia gravis score: Assessment of responsiveness and longitudinal validity , 2005, Neurology.

[42]  D B Sanders,et al.  Myasthenia gravis: recommendations for clinical research standards. Task Force of the Medical Scientific Advisory Board of the Myasthenia Gravis Foundation of America. , 2000, Neurology.

[43]  R. Pirskanen,et al.  Polymorphisms in IL-1β and IL-1 receptor antagonist genes are associated with myasthenia gravis , 1998, Journal of Neuroimmunology.

[44]  K. Takatsu.  Cytokines Involved in B-Cell Differentiation and Their Sites of Action , 1997, Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine.

[45]  D. Perl,et al.  Acid sphingomyelinase deficient mice: a model of types A and B Niemann–Pick disease , 1995, Nature Genetics.

[46]  A. Ochi,et al.  Sphingomyelin‐ceramide turnover in CD28 costimulatory signaling , 1995, European journal of immunology.

[47]  M. Peuchmaur,et al.  In situ production of interleukins in hyperplastic thymus from myasthenia gravis patients. , 1991, Human pathology.

[48]  D. Elmqvist.  Myasthenia Gravis , 1975, The Lancet.

[49]  Sabrin H Albeituni,et al.  Roles of Ceramides and Other Sphingolipids in Immune Cell Function and Inflammation. , 2019, Advances in experimental medicine and biology.

[50]  M. Parnham,et al.  Ceramides as Novel Disease Biomarkers. , 2019, Trends in molecular medicine.

[51]  B. Ogretmen,et al.  Mechanisms of Ceramide-Dependent Cancer Cell Death. , 2018, Advances in cancer research.

[52]  A. Gomez-Muñoz,et al.  Control of inflammatory responses by ceramide, sphingosine 1-phosphate and ceramide 1-phosphate. , 2016, Progress in lipid research.

[53]  R. DeFronzo,et al.  Ceramide content is increased in skeletal muscle from obese insulin-resistant humans. , 2004, Diabetes.