RIPK3 modulates growth factor receptor expression in endothelial cells to support angiogenesis

Angiogenesis. 2021 Aug;24(3):519-531. doi: 10.1007/s10456-020-09763-5. Epub 2021 Jan 15.

Abstract

Receptor-interacting protein kinase 3 (RIPK3) is a multifunctional intracellular protein that was first recognized as an important component of the necroptosis programmed cell death pathway. RIPK3 is also highly expressed in non-necroptotic murine embryonic endothelial cells (ECs) during vascular development, indicating its potential contribution to angiogenesis. To test this hypothesis, we generated mice lacking endothelial RIPK3 and found non-lethal embryonic and perinatal angiogenesis defects in multiple vascular beds. Our in vitro data indicate that RIPK3 supports angiogenesis by regulating growth factor receptor degradation in ECs. We found that RIPK3 interacted with the membrane trafficking protein myoferlin to sustain expression of vascular endothelial growth factor receptor 2 (VEGFR2) in cultured ECs following vascular endothelial growth factor A (VEGFA) stimulation. Restoration of myoferlin, which was diminished after RIPK3 knockdown, rescued decreased VEGFR2 expression and vascular sprouting in RIPK3-deficient ECs after VEGFA treatment. In addition, we found that RIPK3 modulated expression of genes involved in endothelial identity by inhibiting ERK signaling independently of growth factor receptor turnover. Altogether, our data reveal unexpected non-necroptotic roles for RIPK3 in ECs and evidence that RIPK3 promotes developmental angiogenesis in vivo.

Keywords: ERK; Myoferlin; VEGFR2.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Endothelial Cells / metabolism*
  • Gene Expression Regulation*
  • MAP Kinase Signaling System*
  • Mice
  • Mice, Transgenic
  • Neovascularization, Physiologic*
  • Receptor-Interacting Protein Serine-Threonine Kinases / genetics
  • Receptor-Interacting Protein Serine-Threonine Kinases / metabolism*
  • Vascular Endothelial Growth Factor A / genetics
  • Vascular Endothelial Growth Factor A / metabolism
  • Vascular Endothelial Growth Factor Receptor-2 / biosynthesis*
  • Vascular Endothelial Growth Factor Receptor-2 / genetics

Substances

  • Vascular Endothelial Growth Factor A
  • vascular endothelial growth factor A, mouse
  • Kdr protein, mouse
  • Vascular Endothelial Growth Factor Receptor-2
  • Receptor-Interacting Protein Serine-Threonine Kinases
  • Ripk3 protein, mouse