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Proteomic analysis of the inflammatory response of endothelial cells

In collaboration with the team of J-P Girard at IPBS, we use large-scale proteomic analysis associated with label-free quantification to study the pathways and processes activated in endothelial cells (ECs) under inflammatory conditions. 
More than 10 years ago, we developed original bioinformatics tools to extract peptide signals from raw MS files, measure their intensity, and perform quantitative analysis of protein expression in large-scale studies. Using such methods, we could extensively map the ECs proteome, and provided the most complete proteomic characterization to date of the human EC response to several inflammatory cytokines, such as TNFα, IFNgamma and IL1β (1,2).
We keep on applying such a global proteomic approaches, for example to analyze IL-33 function in endothelial cells. IL-33 is a nuclear cytokine from the IL-1 family that plays important roles in health and disease. We compared the extracellular and intracellular roles of IL-33 inprimary human endothelial cells, and found that exogenous extracellular IL-33 acts as a cytokine inducing expression of a distinct set of proteins associated with inflammatory responses in endothelial cells, but not as a nuclear factor (3) .

Capture1.PNGProteomic analysis of the inflammatory response of endothelial cells upon TNFα/IFNgamma stimulation.
Large-scale proteomics enabled us to perform an in-depth profiling of protein expression in endothelial cells, and to characterize the cellular response to stimulation with various cytokines.

Selected publications

  1. Bouyssie, Gonzalez de Peredo et al. Mascot file parsing and quantification (MFPaQ), a new software to parse, validate, and quantify proteomics data generated by ICAT and SILAC mass spectrometric analyses: application to the proteomics study of membrane proteins from primary human endothelial cells. Molecular & cellular proteomics , 2007 https://doi.org/10.1074/mcp.t600069-mcp200
  2. Gautier et al. Label-free quantification and shotgun analysis of complex proteomes by one-dimensional SDS-PAGE/NanoLC-MS: evaluation for the large scale analysis of inflammatory human endothelial cells. Molecular & cellular proteomics, 2012 https://doi.org/10.1074/mcp.m111.015230
  3. Gautier et al. Extracellular IL-33 cytokine, but not endogenous nuclear IL-33, regulates protein expression in endothelial cells. Sci Rep, 2016 https://doi.org/10.1038/srep34255