Lipidome profiling of human and equine neutrophil-derived extracellular vesicles and their potential contribution to the ensemble of synovial fluid-derived extracellular vesicles during joint inflammation

Author:

Varela LauraORCID,Mol Sanne,Taanman-Kueter Esther W.,Ryan Sarah E.,Taams Leonie S.ORCID,de Jong EstherORCID,van Weeren P. RenéORCID,van de Lest Chris H.A.ORCID,Wauben Marca H.M.ORCID

Abstract

ABSTRACTThe molecular signature of cell-derived extracellular vesicles (EVs) from synovial fluid (SF) offers valuable insights into the cells and molecular processes associated with joint disorders and can be exploited to define biomarkers. The signature of EVs is determined by cargo molecules and the lesser-studied lipid bilayer. We here investigated the lipidome of SF-EVs in inflamed joints derived from Rheumatoid Arthritis (RA) and Spondyloarthritis (SpA) patients, two autoimmune-driven joint diseases, and compared these signatures to the lipid profile of equine SF-EVs obtained during induced acute synovitis. Since neutrophils are primary SF-infiltrating cells during these inflammatory joint diseases, we also analyzed how inflammatory stimuli alter the lipidomic profile of human and equine neutrophil-derived EVs (nEVs)in vitroand how these signatures relate to the lipidome signatures of SF-EVs from inflamed joints. We identified neutrophil stimulation-intensity dependent changes in the lipidomic profile of nEVs with elevated presence of dihexosylceramide (lactosylceramide), phosphatidylserine, and phosphatidylethanolamine ether-linked lipid classes in human nEVs upon full neutrophil activation. In horses, levels of monohexosylceramide (glucosylceramide) increased instead of dihexosylceramide, indicating species-specific differences. The lipid profiles of RA and SpA SF-EVs were relatively similar and showed a relative resemblance with stimulated human nEVs. Similarly, the lipidome of equine synovitis-derived SF-EVs closer resembled the one of stimulated equine nEVs. Hence, lipidome profiling can provide insights into the contribution of nEVs to the heterogeneous pool of SF-EVs, deepening our understanding of inflammatory joint diseases and revealing molecular changes in joint homeostasis, which can lead to the development of more precise disease diagnosis and treatment strategies.

Publisher

Cold Spring Harbor Laboratory

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