Targeting autocrine HB-EGF signaling with specific ADAM12 inhibition using recombinant ADAM12 prodomain

Sci Rep. 2015 Oct 19;5:15150. doi: 10.1038/srep15150.

Abstract

Dysregulation of ErbB-family signaling underlies numerous pathologies and has been therapeutically targeted through inhibiting ErbB-receptors themselves or their cognate ligands. For the latter, "decoy" antibodies have been developed to sequester ligands including heparin-binding epidermal growth factor (HB-EGF); however, demonstrating sufficient efficacy has been difficult. Here, we hypothesized that this strategy depends on properties such as ligand-receptor binding affinity, which varies widely across the known ErbB-family ligands. Guided by computational modeling, we found that high-affinity ligands such as HB-EGF are more difficult to target with decoy antibodies compared to low-affinity ligands such as amphiregulin (AREG). To address this issue, we developed an alternative method for inhibiting HB-EGF activity by targeting its cleavage from the cell surface. In a model of the invasive disease endometriosis, we identified A Disintegrin and Metalloproteinase 12 (ADAM12) as a protease implicated in HB-EGF shedding. We designed a specific inhibitor of ADAM12 based on its recombinant prodomain (PA12), which selectively inhibits ADAM12 but not ADAM10 or ADAM17. In endometriotic cells, PA12 significantly reduced HB-EGF shedding and resultant cellular migration. Overall, specific inhibition of ligand shedding represents a possible alternative to decoy antibodies, especially for ligands such as HB-EGF that exhibit high binding affinity and localized signaling.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • ADAM Proteins / antagonists & inhibitors*
  • ADAM Proteins / genetics
  • ADAM Proteins / metabolism
  • ADAM12 Protein
  • Amphiregulin / pharmacology
  • Antibodies, Monoclonal / pharmacology
  • Autocrine Communication*
  • Cell Line
  • Cell Movement / drug effects
  • Endometriosis / genetics
  • Endometriosis / metabolism
  • Endometrium / cytology
  • Endometrium / drug effects
  • Endometrium / metabolism
  • Enzyme Inhibitors / pharmacology*
  • Female
  • Heparin-binding EGF-like Growth Factor / metabolism*
  • Humans
  • Ligands
  • Membrane Proteins / antagonists & inhibitors*
  • Membrane Proteins / genetics
  • Membrane Proteins / metabolism
  • Models, Biological
  • Protein Binding
  • Recombinant Proteins / pharmacology*
  • Signal Transduction / drug effects*

Substances

  • Amphiregulin
  • Antibodies, Monoclonal
  • Enzyme Inhibitors
  • Heparin-binding EGF-like Growth Factor
  • Ligands
  • Membrane Proteins
  • Recombinant Proteins
  • ADAM Proteins
  • ADAM12 Protein
  • ADAM12 protein, human