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GIT1 mediates VEGF-induced podosome formation in endothelial cells: critical role for PLCgamma

Jing Wang1, Yoji Taba, Jinjiang Pang

  • 1Aab Cardiovascular Research Institute, University of Rochester School of Medicine and Dentistry, Rochester, NY 14642, USA.

Arteriosclerosis, Thrombosis, and Vascular Biology
|November 22, 2008
PubMed
Abstract

Insights

G protein-coupled receptor (GPCR) kinase-interacting protein 1 (GIT1) is crucial for vascular endothelial growth factor (VEGF)-induced endothelial cell migration and podosome formation. GIT1 mediates matrix degradation and cell migration by regulating podosome dynamics via phospholipase C-gamma (PLCgamma).

Area of Science:

  • Endothelial cell biology
  • Molecular signaling pathways
  • Angiogenesis research

Background:

  • Tyrosine kinase receptors (TKRs) like VEGF are vital for endothelial cell (EC) functions including survival, migration, and angiogenesis.
  • Podosomes, actin-rich structures, are implicated in EC migration, tissue invasion, and matrix remodeling during angiogenesis.
  • G protein-coupled receptor (GPCR) kinase-interacting protein 1 (GIT1) phosphorylation by c-Src is linked to TKR signaling and phospholipase C-gamma (PLCgamma) activation.

Purpose of the Study:

  • To investigate the role of GIT1 in vascular endothelial growth factor (VEGF)-induced podosome formation and endothelial cell (EC) migration.
  • To elucidate the signaling pathway involving GIT1, c-Src, and PLCgamma in VEGF-driven EC functions.

Main Methods:

  • Exposure of endothelial cells (ECs) to VEGF and assessment of GIT1 colocalization with podosomes.
  • Depletion of GIT1 using siRNA to evaluate its impact on podosome formation, MMP activation, and EC migration.
  • Inhibition of PLCgamma and Src signaling pathways, and expression of dominant-negative small GTPases to dissect the molecular mechanisms.
  • Detection of matrix metalloproteinases (MMPs) at podosome sites and assessment of extracellular matrix (ECM) degradation.

Main Results:

  • VEGF stimulation induced GIT1 colocalization with podosomes in ECs.
  • GIT1 depletion significantly reduced VEGF-induced podosome formation, MMP activation (MMP2, MT-MMP1), ECM degradation, and EC migration.
  • VEGF-induced podosome formation was dependent on Src, GIT1, PLCgamma, and small GTPases.
  • PLCgamma was localized with podosomes and inhibition of PLCgamma or Src significantly impaired podosome formation.

Conclusions:

  • GIT1 is an essential mediator of VEGF-induced podosome formation and EC migration.
  • The signaling pathway involves GIT1, Src, and PLCgamma, highlighting GIT1's critical role in angiogenesis.
  • GIT1 regulates VEGF-induced matrix metalloproteinase activation and extracellular matrix degradation through podosome formation.

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