Early Intervention by Targeting inhibitors of Multiple Signal Transduction Pathways in LPS induced Human PBMCs

Abstract

Lipopolysaccharide (LPS) is an endotoxin that induces the secretion of pro‐inflammatory cytokines in many cells as well as septic shock in humans. This project is aimed at developing a rapid diagnostic method to combat the effects of LPS by utilizing signal transduction pathways induced by LPS in Peripheral Blood Mononuclear Cells (PBMC) as a prototype module. A set of LPS induced genes were identified through micro array analysis and three common signaling pathways were found (p38, JNK, and MAPK). Further investigation of the pathways led to the identification of common components to multiple pathways that have the potential to serve as diagnostic markers. After inhibiting of three pathways by specific inhibitors 4‐[4‐(4‐fluorophenyl)‐2‐[4‐(methylsulfinyl)phenyl]‐1H‐imidazol‐5‐yl]‐pyridine, monohydrochloridem (SB203580), anthrax[1,9‐cd]pyrazol‐6(2H)‐one (SP600125), and 2‐(2‐Amino‐3‐methoxyphenyl)‐4H‐1‐benzopyran‐4‐one (PD98059) we analyzed a few targets by evaluating the time dependent gene expression pattern and then by investigating the impact on components that are indirectly associated with the targets. We will further evaluate the effect of the specific inhibitors by correlating the observed gene expression pattern to the protein expression pattern using ELISA. We believe the study will allow us to better understand the complex interactions of multiple signal transduction pathways induced by LPS.