FGD5 Regulates VEGF Receptor-2 Coupling to PI3 Kinase and Receptor Recycling

Arterioscler Thromb Vasc Biol. 2017 Dec;37(12):2301-2310. doi: 10.1161/ATVBAHA.117.309978. Epub 2017 Oct 19.

Abstract

Objective: VEGF (vascular endothelial growth factor-A) signaling to the endothelial cell (EC) through VEGFR2 (VEGF receptor-2) is the principal cue driving new blood vessel formation. FGD5 (faciogenital dysplasia-5)-a Rho-family guanine nucleotide exchange factor-is selectively expressed in EC. Deficiency of FGD5 is embryonically lethal in mice and perturbs angiogenesis and VEGF signal transduction. However, the mechanism of FGD5 regulation of VEGF signaling is poorly understood.

Approach and results: Angiogenic sprouting and EC cytoskeletal remodeling were evaluated in a 3-dimensional in vitro model. We examined the subcellular localization of FGD5 and VEGFR2 in EC by immunofluorescent staining and studied the association by immunoprecipitation. FGD5 deficiency reduced the number of angiogenic sprouts and tip cell filopodia by ≈80% and ≈70%, respectively. These defects were accompanied by downregulation of the expression of tip cell-specific markers. FGD5 inactivation led to a decrease in EC migration and early protrusion (lamellipodia) formation. In resting and VEGF-stimulated EC, FGD5 forms a complex with VEGFR2 and was enriched at the leading edge of the cell and among endosomes. FGD5 loss reduced mTORC2 (mammalian target of rapamycin complex-2)/Akt-dependent cortactin activation downstream of VEGFR2 but did not alter VEGFR2 plasma membrane expression, Y1175 phosphorylation, or endocytosis. However, FGD5 loss decreased endosomal VEGFR2 coupling to phosphoinositide-3 kinase and diverted VEGFR2 to lysosomal degradation.

Conclusions: FGD5 regulates VEGFR2 retention in recycling endosomes and coupling to PI3 (phosphoinositide-3) kinase/mTORC2-dependent cytoskeletal remodeling.

Keywords: downregulation; endosomes; endothelium; guanine nucleotide exchange factors; signal transduction; vascular endothelial growth factor receptor-2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Movement
  • Cell Proliferation
  • Cells, Cultured
  • Cortactin / metabolism
  • Cytoskeleton / enzymology
  • Endosomes / enzymology
  • Guanine Nucleotide Exchange Factors / genetics
  • Guanine Nucleotide Exchange Factors / metabolism*
  • Human Umbilical Vein Endothelial Cells / enzymology*
  • Humans
  • Lysosomes / enzymology
  • Mechanistic Target of Rapamycin Complex 2 / metabolism
  • Neovascularization, Physiologic*
  • Phosphatidylinositol 3-Kinase / metabolism*
  • Protein Binding
  • Proteolysis
  • Proto-Oncogene Proteins c-akt / metabolism
  • Pseudopodia / enzymology
  • RNA Interference
  • Signal Transduction
  • Time Factors
  • Transfection
  • Vascular Endothelial Growth Factor Receptor-2 / genetics
  • Vascular Endothelial Growth Factor Receptor-2 / metabolism*

Substances

  • CTTN protein, human
  • Cortactin
  • FGD5 protein, human
  • Guanine Nucleotide Exchange Factors
  • Phosphatidylinositol 3-Kinase
  • KDR protein, human
  • Vascular Endothelial Growth Factor Receptor-2
  • Mechanistic Target of Rapamycin Complex 2
  • Proto-Oncogene Proteins c-akt

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