Endothelium-Derived Extracellular Vesicles Expressing Intercellular Adhesion Molecules Reflect Endothelial Permeability and Sepsis Severity

• Published in: Anesthesia and analgesia Vol. 139; no. 2; p. 385
• Main Authors: Takei, Yusuke, Yamada, Mitsuhiro, Saito, Koji, Kameyama, Yoshinobu, Aihara, Takanori, Iwasaki, Yudai, Murakami, Toru, Kaiho, Yu, Ohkoshi, Akira, Konno, Daisuke, Shiga, Takuya, Takahashi, Kazuhiro, Ikumi, Saori, Toyama, Hiroaki, Ejima, Yutaka, Yamauchi, Masanori
• Format: Journal Article
• Language: English
• Published: United States 01.08.2024
• Subjects: Aged; Angiopoietin-1 - metabolism; Antigens, CD - metabolism; Biomarkers - blood; Biomarkers - metabolism; Cadherins - metabolism; Capillary Permeability; Cells, Cultured; Cross-Sectional Studies; Endothelium, Vascular - metabolism; Epoprostenol - metabolism; Extracellular Vesicles - metabolism; Female; Human Umbilical Vein Endothelial Cells - metabolism; Humans; Male; Middle Aged; Platelet Endothelial Cell Adhesion Molecule-1 - metabolism; Prospective Studies; Sepsis - metabolism; Severity of Illness Index; Tumor Necrosis Factor-alpha - metabolism; Tumor Necrosis Factor-alpha - pharmacology; Vascular Endothelial Growth Factor A - metabolism
• ISSN: 1526-7598
• DOI: 10.1213/ANE.0000000000006988

Currently, clinical indicators for evaluating endothelial permeability in sepsis are unavailable. Endothelium-derived extracellular vesicles (EDEVs) are emerging as biomarkers of endothelial injury. Platelet endothelial cell adhesion molecule (PECAM) and vascular endothelial (VE)-cadherin are constitutively expressed endothelial intercellular adhesion molecules that regulate intercellular adhesion and permeability. Herein, we investigated the possible association between EDEVs expressing intercellular adhesion molecules (PECAM+ or VE-cadherin+ EDEVs) and endothelial permeability and sepsis severity. Human umbilical vein endothelial cells (HUVECs) were stimulated with tumor necrosis factor alpha (TNF-α) directly or after pretreatment with permeability-modifying reagents such as angiopoietin-1, prostacyclin, or vascular endothelial growth factor (VEGF) to alter TNF-α-induced endothelial hyperpermeability. Endothelial permeability was measured using the dextran assay or transendothelial electrical resistance. Additionally, a prospective cross-sectional observational study was conducted to analyze circulating EDEV levels in patients with sepsis. EDEVs were examined in HUVEC culture supernatants or patient plasma (nonsepsis, n = 30; sepsis, n = 30; septic shock, n = 42) using flow cytometry. The Wilcoxon rank-sum test was used for comparisons between 2 groups. Comparisons among 3 or more groups were performed using the Steel-Dwass test. Spearman's test was used for correlation analysis. Statistical significance was set at P < .05. TNF-α stimulation of HUVECs significantly increased EDEV release and endothelial permeability. Pretreatment with angiopoietin-1 or prostacyclin suppressed the TNF-α-induced increase in endothelial permeability and inhibited the release of PECAM+ and VE-cadherin+ EDEVs. In contrast, pretreatment with VEGF increased TNF-α-induced endothelial permeability and the release of PECAM+ and VE-cadherin+ EDEVs. However, pretreatment with permeability-modifying reagents did not affect the release of EDEVs expressing inflammatory stimulus-inducible endothelial adhesion molecules such as E-selectin, intracellular adhesion molecule-1, or vascular cell adhesion molecule-1. The number of PECAM+ EDEVs on admission in the septic-shock group (232 [124, 590]/μL) was significantly higher (P = .043) than that in the sepsis group (138 [77,267]/μL), with an average treatment effect of 98/μL (95% confidence interval [CI], 2-270/μL), and the number of VE-cadherin+ EDEVs in the septic-shock group (173 [76,339]/μL) was also significantly higher (P = .004) than that in the sepsis group (81 [42,159]/μL), with an average treatment effect (ATE) of 79/μL (95% CI, 19-171/μL); these EDEV levels remained elevated until day 5. EDEVs expressing intercellular adhesion molecules (PECAM+ or VE-cadherin+ EDEVs) may reflect increased endothelial permeability and could be valuable diagnostic and prognostic markers for sepsis.