Proteome profiling in IL-1β and VEGF-activated human umbilical vein endothelial cells delineates the interlink between inflammation and angiogenesis

PLoS One. 2017 Jun 15;12(6):e0179065. doi: 10.1371/journal.pone.0179065. eCollection 2017.

Abstract

Endothelial cells represent major effectors in inflammation and angiogenesis, processes that drive a multitude of pathological states such as atherosclerosis and cancer. Both inflammation and angiogenesis are interconnected with each other in the sense that many pro-inflammatory proteins possess proangiogenic properties and vice versa. To elucidate this interplay further, we present a comparative proteome study of inflammatory and angiogenic activated endothelial cells. HUVEC were stimulated with interleukin 1-β and VEGF, respectively. Cultured primary cells were fractionated into secreted, cytoplasmic and nuclear protein fractions and processed for subsequent LC-MS/MS analysis. Obtained protein profiles were filtered for fraction-specific proteins to address potential cross fractional contamination, subjected to comparative computational biology analysis (GO-Term enrichment analysis, weighted gene co-expression analysis) and compared to published mRNA profiles of IL-1β respectively VEGF stimulated HUVEC. GO Term enrichment analysis and comparative pathway analysis revealed features such as NOD and NfkB signaling for inflammatory activated HUVEC and VEGF and ErB signaling for VEGF-activated HUVEC with potential crosstalk via map kinases MAP2K2. Weighted protein co-expression network analysis revealed several potential hub genes so far not associated with driver function in inflammation or angiogenesis such as HSPG2, ANXA3, and GPI. "Classical" inflammation or angiogenesis markers such as IL6, CXCL8 or CST1 were found in a less central position within the co-expression networks. In conclusion, this study reports a framework for the computational biology based analysis of proteomics data applied to cytoplasmic, nucleic and extracellular fractions of quiescent, inflammatory and angiogenic activated HUVEC. Novel potential hub genes relevant for these processes were successfully identified.

MeSH terms

  • Gene Expression Profiling
  • Gene Expression Regulation / drug effects*
  • Human Umbilical Vein Endothelial Cells
  • Humans
  • Inflammation / chemically induced
  • Inflammation / metabolism
  • Interleukin-1beta / pharmacology*
  • Neovascularization, Physiologic / drug effects*
  • Proteome / biosynthesis*
  • Proteomics
  • Vascular Endothelial Growth Factor A / pharmacology*

Substances

  • IL1B protein, human
  • Interleukin-1beta
  • Proteome
  • VEGFA protein, human
  • Vascular Endothelial Growth Factor A

Grants and funding

The ScienceConsult – DI Thomas Mohr KG and Evercyte GmbH provided support in the form of income for authors TM (ScienceConsult – DI Thomas Mohr KG) and JG (Evercyte GmbH), but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of these authors are articulated in the ‘author contributions’ section.