Documents

DOI

  • Александр Степанов
  • Дарья Шишкова
  • Виктория Маркова
  • Юлия Маркова
  • Алексей Фролов
  • Анастасия Лазебная
  • Екатерина Ощепкова
  • Дарья Переплетчикова
  • Дарья Смирнова
  • Любовь Басович
  • Егор Алексеевич Репкин
  • Антон Кутихин

Calciprotein particles (CPPs) are essential circulating scavengers of excessive Ca 2+ and PO 4 3- ions, representing a vehicle that removes them from the human body and precludes extraskeletal calcification. Having been internalised by endothelial cells (ECs), CPPs induce their dysfunction, which is accompanied by a remarkable molecular reconfiguration, although little is known about this process's extracellular signatures. Here, we applied ultra-high performance liquid chromatography-tandem mass spectrometry to perform a secretome-wide profiling of the cell culture supernatant from primary human coronary artery ECs (HCAECs) and internal thoracic artery ECs (HITAECs) treated with primary CPPs (CPP-P), secondary CPPs (CPP-S), magnesiprotein particles (MPPs), or Ca 2+/Mg 2+-free Dulbecco's phosphate-buffered saline (DPBS) for 24 h. Incubation with CPP-P/CPP-S significantly altered the profiles of secreted proteins, delineating physiological and pathological endothelial secretomes. Neither pathway enrichment analysis nor the interrogation of protein-protein interactions detected extracellular matrix- and basement membrane-related molecular terms in the protein datasets from CPP-P/CPP-S-treated ECs. Both proteomic profiling and enzyme-linked immunosorbent assay identified an increased level of protectin (CD59) and reduced levels of osteonectin (SPARC), perlecan (HSPG2), and fibronectin (FN1) in the cell culture supernatant upon CPP-P/CPP-S treatment. Elevated soluble CD59 and decreased release of basement membrane components might be considered as potential signs of dysfunctional endothelium.

Original languageEnglish
Article number11382
JournalInternational Journal of Molecular Sciences
Volume25
Issue number21
DOIs
StatePublished - 23 Oct 2024

    Research areas

  • Basement Membrane/metabolism, Cells, Cultured, Coronary Vessels/metabolism, Down-Regulation/drug effects, Endothelial Cells/metabolism, Fibronectins/metabolism, Heparan Sulfate Proteoglycans/metabolism, Humans, Osteonectin/metabolism, Proteome/metabolism, Proteomics/methods, Secretome/metabolism, Tandem Mass Spectrometry, endothelial cells, basement membrane, proteomic profiling, bioinformatic analysis, endothelial dysfunction, endothelial secretome, calcium stress, CD59, calciprotein particles, extracellular matrix

ID: 126354091