Phenotypic and proteomic analysis of plasma extracellular vesicles highlights them as potential biomarkers of primary Sjögren syndrome

Sjögren syndrome (SjS) is an autoimmune disease characterized by the destruction of the exocrine gland epithelia, causing a dryness of mucosa called sicca symptoms, and whose main life-threatening complication is lymphoma. There is a need for new biomarkers in this disease, notably diagnostic biomarkers for patients with genuine sicca symptoms that do not meet current criteria, and prognostic biomarkers for patients at risk of lymphoma. Plasma extracellular vesicles (EVs) are promising biomarker candidates in several diseases, but their potential has not yet been explored in SjS. In this proof-of-concept study, we characterized EVs from primary SjS patients (pSS, n=12) at the phenotypic and proteomic levels, compared to EVs from healthy donor (HD, n=8) and systemic lupus erythematosus patients (SLE, n=12). Specific plasma EVs subpopulations, derived from neutrophils, endothelial, and epithelial cells, were found increased in pSS. We also identified a pSS proteomic signature in plasma EVs, including neutrophil-, epithelial-, and endothelial-related proteins, such as integrin alpha M (ITGAM), olfactomedin-4 (OLFM4), Ras-related protein RAB10, and CD36. Overall, our results support the relevance of plasma EVs as biomarkers in SjS.

[1]  R. Jonsson,et al.  Sjögren syndrome , 2016, Nature Reviews Disease Primers.

[2]  E. Buzás.  The roles of extracellular vesicles in the immune system , 2022, Nature Reviews Immunology.

[3]  M. Castelino,et al.  Multi-Omic Biomarkers for Patient Stratification in Sjogren’s Syndrome—A Review of the Literature , 2022, Biomedicines.

[4]  Qing-jun Wu,et al.  The potential roles of type I interferon activated neutrophils and neutrophil extracellular traps (NETs) in the pathogenesis of primary Sjögren’s syndrome , 2022, Arthritis Research & Therapy.

[5]  F. Dignat-George,et al.  A new strategy to count and sort neutrophil‐derived extracellular vesicles: Validation in infectious disorders , 2022, Journal of extracellular vesicles.

[6]  A. Handberg,et al.  Lipid‐based strategies used to identify extracellular vesicles in flow cytometry can be confounded by lipoproteins: Evaluations of annexin V, lactadherin, and detergent lysis , 2022, Journal of extracellular vesicles.

[7]  B. Thiede,et al.  Proteomic Profiling of Saliva and Tears in Radiated Head and Neck Cancer Patients as Compared to Primary Sjögren’s Syndrome Patients , 2022, International journal of molecular sciences.

[8]  G. Jay,et al.  Proteomics Analysis of Tears and Saliva From Sjogren’s Syndrome Patients , 2021, Frontiers in Pharmacology.

[9]  S. Rocchiccioli,et al.  Characterization of Extracellular Vesicle Cargo in Sjögren’s Syndrome through a SWATH-MS Proteomics Approach , 2021, International journal of molecular sciences.

[10]  A. Tzioufas,et al.  New frontiers in precision medicine for Sjogren’s syndrome , 2021, Expert review of clinical immunology.

[11]  D. Fotiadis,et al.  Sjögren’s Syndrome: The Clinical Spectrum of Male Patients , 2020, Journal of clinical medicine.

[12]  David Gray,et al.  In Human Autoimmunity, a Substantial Component of the B Cell Repertoire Consists of Polyclonal, Barely Mutated IgG+ve B Cells , 2020, Frontiers in Immunology.

[13]  X. Mariette,et al.  EULAR recommendations for the management of Sjögren’s syndrome with topical and systemic therapies , 2019, Annals of the rheumatic diseases.

[14]  B. Thiede,et al.  Proteomic and histopathological characterisation of sicca subjects and primary Sjögren’s syndrome patients reveals promising tear, saliva and extracellular vesicle disease biomarkers , 2019, Arthritis Research & Therapy.

[15]  T. Andresen,et al.  What is the blood concentration of extracellular vesicles? Implications for the use of extracellular vesicles as blood-borne biomarkers of cancer. , 2019, Biochimica et biophysica acta. Reviews on cancer.

[16]  Jing Xu,et al.  Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines , 2018, Journal of Extracellular Vesicles.

[17]  B. Tang,et al.  Rab 10—a traffic controller in multiple cellular pathways and locations , 2018, Journal of cellular physiology.

[18]  L. Criswell,et al.  Primary Sjögren's Syndrome , 2018, The New England journal of medicine.

[19]  Graça Raposo,et al.  Shedding light on the cell biology of extracellular vesicles , 2018, Nature Reviews Molecular Cell Biology.

[20]  B. Thiede,et al.  Identification of potential saliva and tear biomarkers in primary Sjögren’s syndrome, utilising the extraction of extracellular vesicles and proteomics analysis , 2017, Arthritis Research & Therapy.

[21]  M. Ranson,et al.  PZP and PAI-2: Structurally-diverse, functionally similar pregnancy proteins? , 2016, The international journal of biochemistry & cell biology.

[22]  William Stafford Noble,et al.  Fast and Accurate Protein False Discovery Rates on Large-Scale Proteomics Data Sets with Percolator 3.0 , 2016, Journal of The American Society for Mass Spectrometry.

[23]  Wenli Liu,et al.  Olfactomedin 4 expression and functions in innate immunity, inflammation, and cancer , 2016, Cancer and Metastasis Reviews.

[24]  Michael W. Jackson,et al.  Secreted autoantibody repertoires in Sjögren's syndrome and systemic lupus erythematosus: A proteomic approach. , 2016, Autoimmunity reviews.

[25]  R. Gerli,et al.  Characterization of circulating endothelial microparticles and endothelial progenitor cells in primary Sjögren's syndrome: new markers of chronic endothelial damage? , 2015, Rheumatology.

[26]  Marco Y. Hein,et al.  Accurate Proteome-wide Label-free Quantification by Delayed Normalization and Maximal Peptide Ratio Extraction, Termed MaxLFQ * , 2014, Molecular & Cellular Proteomics.

[27]  Young Mi Park.  CD36, a scavenger receptor implicated in atherosclerosis , 2014, Experimental & Molecular Medicine.

[28]  R. Nieuwland,et al.  Co-isolation of extracellular vesicles and high-density lipoproteins using density gradient ultracentrifugation , 2014, Journal of extracellular vesicles.

[29]  R. Nieuwland,et al.  Single-step isolation of extracellular vesicles by size-exclusion chromatography , 2014, Journal of extracellular vesicles.

[30]  Alexander Schmidt,et al.  Critical assessment of proteome‐wide label‐free absolute abundance estimation strategies , 2013, Proteomics.

[31]  Olivier Langella,et al.  MassChroQ: A versatile tool for mass spectrometry quantification , 2011, Proteomics.

[32]  M. Selbach,et al.  Global quantification of mammalian gene expression control , 2011, Nature.

[33]  G. Illei,et al.  MicroRNA expression profiles as biomarkers of minor salivary gland inflammation and dysfunction in Sjögren's syndrome. , 2011, Arthritis and rheumatism.

[34]  X. Mariette,et al.  Increased levels of circulating microparticles in primary Sjögren's syndrome, systemic lupus erythematosus and rheumatoid arthritis and relation with disease activity , 2009, Arthritis research & therapy.

[35]  Emmanuel Barillot,et al.  myProMS, a web server for management and validation of mass spectrometry‐based proteomic data , 2007, Proteomics.

[36]  M. Gorenstein,et al.  Absolute Quantification of Proteins by LCMSE , 2006, Molecular & Cellular Proteomics.

[37]  H. Moutsopoulos,et al.  Salivary gland epithelial cell exosomes: A source of autoantigenic ribonucleoproteins. , 2005, Arthritis and rheumatism.

[38]  G. Illei,et al.  Exosomes from human saliva as a source of microRNA biomarkers. , 2010, Oral diseases.