Inhibitory effect of nintedanib on VEGF secretion in retinal pigment epithelial cells induced by exposure to a necrotic cell lysate

PLoS One. 2019 Aug 6;14(8):e0218632. doi: 10.1371/journal.pone.0218632. eCollection 2019.

Abstract

Necrosis is a form of cell death that results in rupture of the plasma membrane and the release of cellular contents, and it can give rise to sterile inflammation in the retina and other tissues. The secretion of vascular endothelial growth factor (VEGF) by retinal pigment epithelial (RPE) cells contributes to retinal homeostasis as well as to pathological angiogenesis. We have now examined the effect of a necrotic cell lysate prepared from human RPE cells (NLR) on the release of VEGF by healthy RPE cells. We found that NLR markedly increased the release of VEGF from RPE cells and that this effect was attenuated by nintedanib, a multiple receptor tyrosine kinase inhibitor, whereas it was unaffected by inhibitors of NF-κB signaling or of caspase-1. NLR also induced the phosphorylation of extracellular signal-regulated kinase (Erk) and signal transducer and activator of transcription 3 (Stat3) in a manner sensitive to inhibition by nintedanib, although inhibitors of Erk and Stat3 signaling pathways did not affect NLR-induced VEGF secretion. In addition, nintedanib attenuated the development of choroidal neovascularization in mice. Our results have thus shown that a necrotic lysate of RPE cells induced VEGF secretion from healthy RPE cells and that this effect was mediated by receptor tyrosine kinase signaling. They therefore suggest that VEGF secretion by healthy RPE cells is a potential therapeutic target for retinal diseases associated with sterile inflammation and pathological angiogenesis.

MeSH terms

  • Animals
  • Cell Line
  • Choroidal Neovascularization / drug therapy
  • Choroidal Neovascularization / pathology
  • Extracellular Signal-Regulated MAP Kinases / metabolism
  • Focal Adhesions / drug effects
  • Focal Adhesions / pathology
  • Humans
  • Indoles / pharmacology*
  • Indoles / therapeutic use
  • Mice
  • Necrosis / pathology
  • Protein Kinase Inhibitors / pharmacology*
  • Protein Kinase Inhibitors / therapeutic use
  • Proto-Oncogene Proteins c-akt / metabolism
  • Receptor Protein-Tyrosine Kinases / antagonists & inhibitors
  • Receptor Protein-Tyrosine Kinases / metabolism
  • Retinal Pigment Epithelium / cytology*
  • Retinal Pigment Epithelium / drug effects
  • Retinal Pigment Epithelium / metabolism
  • Retinal Pigment Epithelium / pathology
  • STAT3 Transcription Factor / metabolism
  • Signal Transduction / drug effects
  • Vascular Endothelial Growth Factor A / metabolism*

Substances

  • Indoles
  • Protein Kinase Inhibitors
  • STAT3 Transcription Factor
  • Vascular Endothelial Growth Factor A
  • Receptor Protein-Tyrosine Kinases
  • Proto-Oncogene Proteins c-akt
  • Extracellular Signal-Regulated MAP Kinases
  • nintedanib

Grants and funding

The author(s) received no specific funding for this work.