Key Proteins Involved in Spheroid Formation and Angiogenesis in Endothelial Cells After Long-Term Exposure to Simulated Microgravity

Cell Physiol Biochem. 2018;45(2):429-445. doi: 10.1159/000486920. Epub 2018 Jan 24.

Abstract

Background/aims: Cardiovascular complications are common in astronauts returning from a prolonged spaceflight. These health problems might be driven by complex modulations of gene expression and protein synthesis in endothelial cells (ECs). Studies on the influence of microgravity on phenotype, growth pattern and biological processes of ECs can help to understand these complications.

Methods: We exposed ECs (EA.hy926) to a Random Positioning Machine (RPM). Proteins associated with cell structure, angiogenesis and endothelial dysfunction were investigated in distinct pools of multicellular spheroids (MCS), adherent cells (AD) and tubular structures (TS) formed after a 35-day RPM-exposure.

Results: Combining morphological and molecular approaches, we found AD, MCS and TS with changes in the synthesis and release of proteins involved in three-dimensional growth. Fibronectin and monocyte chemoattractant protein-1 (MCP-1) mRNAs and protein contents were elevated along with an increased secretion of vascular endothelial growth factor (VEGF), interleukin (IL)-6, IL-8, MCP-1, intercellular adhesion molecule 1 (ICAM-1), vascular cell adhesion molecule 1 (VCAM-1), neutrophil gelatinase-associated lipocalin (NGAL) and regulated on activation, normal T cell expressed and secreted (RANTES) proteins in the culture supernatant as determined by multianalyte profiling technology. Together they form a network of interaction.

Conclusions: These results show that a prolonged RPM-exposure of ECs induced TS and MCS formation. The factors VEGF, NGAL, IL-6, IL-8, MCP-1, VCAM-1, ICAM-1, fibronectin and RANTES seem to be affected when gravity is omitted.

Keywords: Angiogenesis; Endothelial cells; Endothelial dysfunction; Microgravity; Multicellular spheroids; Random Positioning Machine; Tube formation.

MeSH terms

  • A549 Cells
  • Cell Adhesion
  • Cell Culture Techniques / instrumentation
  • Cell Fusion
  • Cell Line
  • Chemokine CCL2 / genetics
  • Chemokine CCL2 / metabolism
  • Chemokine CCL5 / analysis
  • Fibronectins / genetics
  • Fibronectins / metabolism
  • Human Umbilical Vein Endothelial Cells
  • Humans
  • Intercellular Adhesion Molecule-1 / metabolism
  • Interleukin-6 / analysis
  • Interleukin-8 / analysis
  • Lipocalin-2 / analysis
  • Neovascularization, Physiologic*
  • Spheroids, Cellular / cytology
  • Spheroids, Cellular / metabolism*
  • Up-Regulation
  • Vascular Cell Adhesion Molecule-1 / metabolism
  • Vascular Endothelial Growth Factor A / metabolism
  • Weightlessness Simulation*

Substances

  • Chemokine CCL2
  • Chemokine CCL5
  • Fibronectins
  • Interleukin-6
  • Interleukin-8
  • Lipocalin-2
  • Vascular Cell Adhesion Molecule-1
  • Vascular Endothelial Growth Factor A
  • Intercellular Adhesion Molecule-1