Identification of ILK as a critical regulator of VEGFR3 signalling and lymphatic vascular growth

EMBO J. 2019 Jan 15;38(2):e99322. doi: 10.15252/embj.201899322. Epub 2018 Dec 5.

Abstract

Vascular endothelial growth factor receptor-3 (VEGFR3) signalling promotes lymphangiogenesis. While there are many reported mechanisms of VEGFR3 activation, there is little understanding of how VEGFR3 signalling is attenuated to prevent lymphatic vascular overgrowth and ensure proper lymph vessel development. Here, we show that endothelial cell-specific depletion of integrin-linked kinase (ILK) in mouse embryos hyper-activates VEGFR3 signalling and leads to overgrowth of the jugular lymph sacs/primordial thoracic ducts, oedema and embryonic lethality. Lymphatic endothelial cell (LEC)-specific deletion of Ilk in adult mice initiates lymphatic vascular expansion in different organs, including cornea, skin and myocardium. Knockdown of ILK in human LECs triggers VEGFR3 tyrosine phosphorylation and proliferation. ILK is further found to impede interactions between VEGFR3 and β1 integrin in vitro and in vivo, and endothelial cell-specific deletion of an Itgb1 allele rescues the excessive lymphatic vascular growth observed upon ILK depletion. Finally, mechanical stimulation disrupts the assembly of ILK and β1 integrin, releasing the integrin to enable its interaction with VEGFR3. Our data suggest that ILK facilitates mechanically regulated VEGFR3 signalling via controlling its interaction with β1 integrin and thus ensures proper development of lymphatic vessels.

Keywords: VEGFR3; integrin‐linked kinase; lymphatic vasculature; mechanical stimulation; β1 integrin.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Proliferation
  • Cells, Cultured
  • Disease Models, Animal
  • Female
  • Humans
  • Integrin beta1 / metabolism*
  • Lymphangiogenesis*
  • Lymphatic Vessels / cytology
  • Lymphatic Vessels / metabolism
  • Mice
  • Phosphorylation
  • Protein-Serine-Threonine Kinases / genetics*
  • Protein-Serine-Threonine Kinases / metabolism*
  • Signal Transduction
  • Vascular Endothelial Growth Factor Receptor-3 / metabolism*

Substances

  • Integrin beta1
  • integrin-linked kinase
  • FLT4 protein, human
  • Vascular Endothelial Growth Factor Receptor-3
  • Protein-Serine-Threonine Kinases