PMID- 30153514 OWN - NLM STAT- MEDLINE DCOM- 20200226 LR - 20200226 IS - 1876-7737 (Electronic) IS - 1874-3919 (Linking) VI - 192 DP - 2019 Feb 10 TI - Integrated proteomic analysis of tumor necrosis factor alpha and interleukin 1beta-induced endothelial inflammation. PG - 89-101 LID - S1874-3919(18)30323-3 [pii] LID - 10.1016/j.jprot.2018.08.011 [doi] AB - The vascular endothelium provides a unique interaction plane for plasma proteins and leukocytes in inflammation. The pro-inflammatory cytokines Tumor Necrosis Factor alpha (TNFalpha) and interleukin 1beta (IL-1beta) have a profound effect on endothelial cells, which includes increased levels of adhesion molecules and a disrupted barrier function. To assess the endothelial response to these cytokines at the protein level, we evaluated changes in the whole proteome, cell surface proteome and phosphoproteome after 24 h of cytokine treatment. The effects of TNFalpha and IL-1beta on endothelial cells were strikingly similar and included changes in proteins not previously associated with endothelial inflammation. Temporal profiling revealed time-dependent proteomic changes, including a limited number of early responsive proteins such as adhesion receptors ICAM1 and SELE. In addition, this approach uncovered a greater number of late responsive proteins, including proteins related to self-antigen peptide presentation, and a transient increase in ferritin. Peptide-based cell surface proteomics revealed extensive changes at the cell surface, which were in agreement with the whole proteome. In addition, site-specific changes within ITGA5 and ICAM1 were detected. Combined, our integrated proteomic data provide detailed information on endothelial inflammation, emphasize the role of the extracellular matrix therein, and include potential targets for therapeutic intervention. SIGNIFICANCE: Pro-inflammatory cytokines induce the expression of cell adhesion molecules in vascular endothelial cells. These molecules mediate the adhesion and migration of immune cells across the vessel wall, which is a key process to resolve infections in the underlying tissue. Dysregulation of endothelial inflammation can contribute to vascular diseases and the vascular endothelium is therefore an attractive target to control inflammation. Current strategies targeting endothelial adhesion molecules, including PECAM, CD99, ICAM1 and VCAM1 do not completely prevent transmigration. To identify additional therapeutic targets, we mapped the endothelial proteome after pro-inflammatory cytokine treatment. In addition to the whole proteome, we assessed the surface proteome to focus on cell adhesion molecules, and the phosphoproteome to uncover protein activation states. Here, we present an integrated overview of affected processes which further improves our understanding of endothelial inflammation and may eventually aid in therapeutic intervention of imbalanced inflammation. CI - Copyright (c) 2018 Elsevier B.V. All rights reserved. FAU - Beguin, Eelke P AU - Beguin EP AD - Department of Molecular and Cellular Hemostasis, Sanquin Research, Amsterdam 1066 CX, the Netherlands. FAU - van den Eshof, Bart L AU - van den Eshof BL AD - Department of Molecular and Cellular Hemostasis, Sanquin Research, Amsterdam 1066 CX, the Netherlands. FAU - Hoogendijk, Arie J AU - Hoogendijk AJ AD - Department of Molecular and Cellular Hemostasis, Sanquin Research, Amsterdam 1066 CX, the Netherlands. FAU - Nota, Benjamin AU - Nota B AD - Department of Research Facilities, Sanquin Research and Landsteiner Laboratory, Amsterdam Medical Center, University of Amsterdam, Amsterdam 1066 CX, the Netherlands. FAU - Mertens, Koen AU - Mertens K AD - Department of Molecular and Cellular Hemostasis, Sanquin Research, Amsterdam 1066 CX, the Netherlands; Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences (UIPS), Utrecht University, Utrecht 3584 CS, the Netherlands. FAU - Meijer, Alexander B AU - Meijer AB AD - Department of Molecular and Cellular Hemostasis, Sanquin Research, Amsterdam 1066 CX, the Netherlands; Department of Biomolecular Mass Spectrometry and Proteomics, Utrecht Institute for Pharmaceutical Sciences (UIPS), Utrecht University, Utrecht 3584 CS, the Netherlands. FAU - van den Biggelaar, Maartje AU - van den Biggelaar M AD - Department of Molecular and Cellular Hemostasis, Sanquin Research, Amsterdam 1066 CX, the Netherlands. Electronic address: m.vandenbiggelaar@sanquin.nl. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20180825 PL - Netherlands TA - J Proteomics JT - Journal of proteomics JID - 101475056 RN - 0 (Cell Adhesion Molecules) RN - 0 (IL1B protein, human) RN - 0 (Interleukin-1beta) RN - 0 (Tumor Necrosis Factor-alpha) SB - IM MH - Cell Adhesion Molecules/*biosynthesis MH - Cells, Cultured MH - Endothelial Cells/*metabolism/pathology MH - Humans MH - Inflammation/chemically induced/metabolism/pathology MH - Interleukin-1beta/*pharmacology MH - Proteomics MH - Tumor Necrosis Factor-alpha/*pharmacology OTO - NOTNLM OT - Cell surface proteomics OT - Endothelium OT - IL-1beta OT - Inflammation OT - Phosphoproteomics OT - Proteomics OT - TNFalpha EDAT- 2018/08/29 06:00 MHDA- 2020/02/27 06:00 CRDT- 2018/08/29 06:00 PHST- 2018/06/12 00:00 [received] PHST- 2018/08/15 00:00 [revised] PHST- 2018/08/23 00:00 [accepted] PHST- 2018/08/29 06:00 [pubmed] PHST- 2020/02/27 06:00 [medline] PHST- 2018/08/29 06:00 [entrez] AID - S1874-3919(18)30323-3 [pii] AID - 10.1016/j.jprot.2018.08.011 [doi] PST - ppublish SO - J Proteomics. 2019 Feb 10;192:89-101. doi: 10.1016/j.jprot.2018.08.011. Epub 2018 Aug 25.