In‐depth proteomics approach of secretome to identify novel biomarker for sepsis in LPS‐stimulated endothelial cells

Abstract

Sepsis and septic shock, which are conditions triggered by infection, occur with high incidence in emergency departments and are among the most common causes of death in hospitalized patients worldwide. Therefore, the identification of sepsis biomarkers for the rapid diagnosis is a major goal for researchers in the field of critical care. Endothelial cells play a pivotal role in orchestrating the inflammatory response triggered by sepsis. In this study, we used proteomics to investigate the secretome of EA.hy926 endothelial cells following lipopolysaccharide (LPS) stimulation with 1 μg/mL LPS for 12 or 24 h. SILAC in cell cultures and an online 2D-LC-MS/MS system were used to analyze the secretome dynamics in response to LPS. We found that 22 of the 77 secreted proteins identified in both the presence and absence of LPS and that 19 of the secreted proteins were quantified more strongly after LPS treatment for 24 h than after treatment for 12 h. By Gene Ontology and KEGG pathway analyses, we found that proteins related to the regulation of the actin cytoskeleton showed the highest secretion response to LPS stimulation. Out of the 19 candidate proteins, we focused on moesin, which is involved in the function of endothelial cells, and confirmed its amount in cellular lysates and media taken from primary human umbilical vein endothelial cells treated with LPS. To our knowledge, this study provides the first in-depth analysis of the LPS-induced secretome in human endothelial cells, and we propose 19 new biomarker candidates for sepsis, including moesin.

The miR-15a affected the development of sepsis and septic shock by suppression BCL-2

“…Sepsis remains a common, expensive, and deadly problem throughout the world 21 . To explore the influencing factors such as miR-15a in the progression of sepsis, the rat sepsis model (sepsis group and septic shock group) was established in current study.…”

Cardiac Remodeling: Endothelial Cells Have More to Say Than Just NO

“…Although the method used in this study simulates the pathophysiology of sepsis rather than cardiac remodeling, many inflammatory pathways are also activated in cardiac remodeling. Interestingly, endotoxins upregulate secretion of some of the proteins present in our index list; e.g., thrombospondin-1 secretion increases 1.2-fold, follistatin-related protein 1 secretion increases 1.2-fold, and connective tissue growth factor increases 1.8-fold (Kwon et al, 2015 ; Table 4 ). In a separate mass-spectrometry study in the same EC line it was shown that atorvastatin decreases protein secretion of thrombospondin-1, thrombospondin-2, and connective tissue growth factor (Brioschi et al, 2013 ).…”

“…Therefore, we searched literature for mass-spectrometry data on the secretome of ECs. Specific data on cardiac microvascular ECs are not available, but mass spectrometry data have been published on the secretome of HUVECs (Tunica et al, 2009 ), endothelial progenitor cells (Hemmen et al, 2011 ), and EA.hy926 ECs (HUVEC hybridoma cell line) (Brioschi et al, 2013 ; Kwon et al, 2015 ).…”

“…A recent study investigated the in vitro response of endothelial responses to endotoxins (Kwon et al, 2015 ). Although the method used in this study simulates the pathophysiology of sepsis rather than cardiac remodeling, many inflammatory pathways are also activated in cardiac remodeling.…”

In Search of a Cure for Sepsis: Taming the Monster in Critical Care Medicine

“…Although their data are comparable to PCT measurement for sepsis diagnosis, they showed that their metabolomics approach was a better predictor of disease outcome. Kwon et al [26 ]used proteomics to analyze the secretome of endothelial cells following LPS stimulation and subsequently identified moesin and 19 new sepsis biomarker candidates. Nakada et al [27 ]used bioinformatics approach to identify a single nucleotide polymorphism on the SVEP1 gene that is associated with increased sepsis mortality.…”