Multi-omics delineation of cytokine-induced endothelial inflammatory states

Vascular endothelial cells (ECs) form a dynamic interface between blood and tissue and play a crucial role in the progression of vascular inflammation. Here, we aim to dissect the system-wide molecular mechanisms of inflammatory endothelial-cytokine responses. Applying an unbiased cytokine library, we determined that TNF alpha and IFN gamma induced the largest EC response resulting in distinct proteomic inflammatory signatures. Notably, combined TNF alpha + IFN gamma stimulation induced an additional synergetic inflammatory signature. We employed a multi-omics approach to dissect these inflammatory states, combining (phospho-) proteome, transcriptome and secretome and found, depending on the stimulus, a wide-array of altered immune-modulating processes, including complement proteins, MHC complexes and distinct secretory cytokines. Synergy resulted in cooperative activation of transcript induction. This resource describes the intricate molecular mechanisms that are at the basis of endothelial inflammation and supports the adaptive immunomodulatory role of the endothelium in host defense and vascular inflammation. Combined TNF alpha + IFN gamma stimulation induces a synergetic inflammatory signature, which is the basis of endothelial inflammation, emphasizing the role of endothelial cell-cytokine interactions in inflammatory pathogenesis and role of endothelial cells in inflammation.