Abstract
Sepsis is an exaggerated immune response upon infection with lipopolysaccharide (LPS) as the main causative agent. LPS-induced activation and apoptosis of endothelial cells (EC) can lead to organ dysfunction and finally organ failure. We previously demonstrated that the first twenty amino acids of the Apurinic/Apyrimidinic Endodeoxyribonuclease 1 (APEX1) are sufficient to inhibit EC apoptosis. To identify genes whose regulation by LPS is affected by this N-terminal APEX1 peptide, EC were transduced with an expression vector for the APEX1 peptide or an empty control vector and treated with LPS. Following RNA deep sequencing, genes upregulated in LPS-treated EC expressing the APEX1 peptide were identified bioinformatically. Selected candidates were validated by semi-quantitative real time PCR, a promising one was Selenoprotein T (SELENOT). For functional analyses, an expression vector for SELENOT was generated. To study the effect of SELENOT expression on LPS-induced EC activation and apoptosis, the SELENOT vector was transfected in EC. Immunostaining showed that SELENOT was expressed and localized in the ER. EC transfected with the SELENOT plasmid showed no activation and reduced apoptosis induced by LPS. SELENOT as well as APEX1(1-20) can protect EC against activation and apoptosis and could provide new therapeutic approaches in the treatment of sepsis.
Highlights
Sepsis can best be described as an overwhelming inflammatory condition, in which the body responds to an infection in a hyperactive, dysregulated way, which in turn results in life-threatening organ dysfunction and eventually septic shock
Primary human endothelial cells (EC) were transduced with either a lentiviral vector leading to moderate expression of Apyrimidinic Endodeoxyribonuclease 1 (APEX1)(1-20)
To identify APEX1(1-20)-specific transcriptome changes in response to LPS, we analyzed which genes were regulated by LPS in the APEX1(1-20) expressing cells, but not in the cells transduced with the empty vector
Summary
Sepsis can best be described as an overwhelming inflammatory condition, in which the body responds to an infection in a hyperactive, dysregulated way, which in turn results in life-threatening organ dysfunction and eventually septic shock. According to an estimate of the World Health Organization (WHO), sepsis affects more than 48 million people every year, potentially leading to 11 million deaths [1]. The basis for the pathophysiological responses in the context of sepsis is multifactorial. Except for the introduction of vasopressor agents 40 years ago, no new therapeutic principle for the treatment of sepsis has been developed until today. Lipopolysaccharide (LPS) is an outer membrane component of Gram-negative bacteria. Most bacterial LPS molecules are thermostable and generate a pro-inflammatory stimulus
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