title: Targeting autocrine HB-EGF signaling with specific ADAM12 inhibition using recombinant ADAM12 prodomain.
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.
authors.affiliation: Massachusetts Institute of Technology, Department of Biological Engineering, 77 Massachusetts Ave., Cambridge, MA 02139.
authors.forename: Miles A
authors.initials: MA
authors.lastname: Miller
authors.affiliation: BioZyme, Inc., 1513 Old White Oak Church Road, Apex, NC 27523.
authors.forename: Marcia L
authors.initials: ML
authors.lastname: Moss
authors.affiliation: National Institutes of Environmental Health Services (NIEHS), 111 TW Alexander Dr. RTP, NC 27709.
authors.forename: Gary
authors.initials: G
authors.lastname: Powell
authors.affiliation: National Institutes of Environmental Health Services (NIEHS), 111 TW Alexander Dr. RTP, NC 27709.
authors.forename: Robert
authors.initials: R
authors.lastname: Petrovich
authors.affiliation: National Institutes of Environmental Health Services (NIEHS), 111 TW Alexander Dr. RTP, NC 27709.
authors.forename: Lori
authors.initials: L
authors.lastname: Edwards
authors.affiliation: Massachusetts Institute of Technology, Department of Biological Engineering, 77 Massachusetts Ave., Cambridge, MA 02139.
authors.forename: Aaron S
authors.initials: AS
authors.lastname: Meyer
authors.affiliation: Massachusetts Institute of Technology, Department of Biological Engineering, 77 Massachusetts Ave., Cambridge, MA 02139.
authors.forename: Linda G
authors.initials: LG
authors.lastname: Griffith
authors.affiliation: Massachusetts Institute of Technology, Department of Biological Engineering, 77 Massachusetts Ave., Cambridge, MA 02139.
authors.forename: Douglas A
authors.initials: DA
authors.lastname: Lauffenburger
mesh_terms: D051722:ADAM Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D000072199:ADAM12 Protein; D066258:Amphiregulin / Q000494:pharmacology; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D019898:Autocrine Communication; D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects; D004715:Endometriosis / Q000235:genetics / Q000378:metabolism; D004717:Endometrium / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005260:Female; D066257:Heparin-binding EGF-like Growth Factor / Q000378:metabolism*; D006801:Humans; D008024:Ligands; D008565:Membrane Proteins / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism; D008954:Models, Biological; D011485:Protein Binding; D011994:Recombinant Proteins / Q000494:pharmacology*; D015398:Signal Transduction / Q000187:drug effects*
chemical_list: D066258:Amphiregulin; D000911:Antibodies, Monoclonal; D004791:Enzyme Inhibitors; D066257:Heparin-binding EGF-like Growth Factor; D008024:Ligands; D008565:Membrane Proteins; D011994:Recombinant Proteins; D051722:ADAM Proteins; D000072199:ADAM12 Protein; C000606480:ADAM12 protein, human