The angiogenic regulator CD13/APN is a transcriptional target of Ras signaling pathways in endothelial morphogenesis

Blood. 2003 Mar 1;101(5):1818-26. doi: 10.1182/blood-2002-05-1422. Epub 2002 Oct 24.

Abstract

Angiogenesis, the formation of new blood vessels, is a critical step for tumor growth and metastasis and an integral component of the pathologic inflammatory response in arthritis and the proliferative retinopathies. The CD13/aminopeptidase N (CD13/APN) metalloprotease is an important regulator of angiogenesis where its expression on activated blood vessels is induced by angiogenic signals. Here, we show that cytokine induction of CD13/APN in endothelial cells is regulated by distinct Ras effector pathways involving Ras/mitogen-activated protein kinase (MAPK) or PI-3K. Signals transduced by activated Ras, Raf, and mitogen-induced extracellular kinase (MEK) stimulate transcription from the CD13/APN proximal promoter. Inhibition of these pathways and extracellular signal-regulated serine/threonine kinase (ERK-2) and PI-3K by expression of dominant-negative proteins or chemical inhibitors prevented induction of CD13/APN transcription in response to basic fibroblast growth factor (bFGF). We show that Ras-induced signal transduction is required for growth factor-induced angiogenesis, because inhibition of downstream mediators of Ras signaling (MEK or PI-3K) abrogated endothelial cell migration, invasion, and morphogenesis in vitro. Reintroduction of CD13/APN, a shared downstream target of these pathways, overrode the suppressive effect of these inhibitors and restored the function of endothelial cells in migration/invasion and capillary morphogenesis assays. Similarly, inhibition of MEK abrogated cell invasion and the formation of endothelial-lined capillaries in vivo, which was effectively rescued by addition of exogenous CD13/APN protein. These studies provide strong evidence that CD13/APN is an important target of Ras signaling in angiogenesis and is a limiting factor in angiogenic progression.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • CD13 Antigens / biosynthesis
  • CD13 Antigens / genetics
  • CD13 Antigens / physiology*
  • Capillaries / ultrastructure
  • Endothelium, Vascular / cytology*
  • Enzyme Induction / drug effects
  • Fibroblast Growth Factor 2 / pharmacology
  • Genes, Dominant
  • Genes, ras
  • Hemangioendothelioma / pathology
  • Humans
  • MAP Kinase Kinase 1
  • MAP Kinase Signaling System
  • Mice
  • Mitogen-Activated Protein Kinase 1 / physiology
  • Mitogen-Activated Protein Kinase Kinases / physiology
  • Morphogenesis
  • Neovascularization, Pathologic / enzymology
  • Neovascularization, Physiologic / drug effects
  • Neovascularization, Physiologic / physiology*
  • Phosphatidylinositol 3-Kinases / physiology
  • Promoter Regions, Genetic
  • Protein Serine-Threonine Kinases / physiology
  • Proto-Oncogene Proteins p21(ras) / physiology*
  • Recombinant Fusion Proteins / physiology
  • Signal Transduction / physiology*
  • Transcription, Genetic* / drug effects
  • Transfection
  • Tumor Cells, Cultured

Substances

  • Recombinant Fusion Proteins
  • Fibroblast Growth Factor 2
  • Protein Serine-Threonine Kinases
  • Mitogen-Activated Protein Kinase 1
  • MAP Kinase Kinase 1
  • MAP2K1 protein, human
  • Map2k1 protein, mouse
  • Mitogen-Activated Protein Kinase Kinases
  • CD13 Antigens
  • HRAS protein, human
  • Proto-Oncogene Proteins p21(ras)