Pertuzumab increases epidermal growth factor receptor down-regulation by counteracting epidermal growth factor receptor-ErbB2 heterodimerization.
Epidermal growth factor receptor (EGFR) and ErbB2 readily form heterodimers when both are expressed in the same cell and the EGFR is activated by one of its ligands. Our data show that such heterodimers are constitutively formed also in a ligand-independent manner on overexpression of EGFR and ErbB2 in porcine aortic endothelial cells. Interestingly, cross-linking experiments showed that incubation with the antibody pertuzumab, which has been shown to bind the dimerization arm of ErbB2, resulted in dissolution of EGFR-ErbB2 heterodimers. Incubation with pertuzumab also increased the amount of EGF-induced EGFR homodimers, and under these conditions, endocytosis of radiolabeled EGF was increased. This increase was significant, although slightly more EGF was internalized in cells expressing EGFR only compared with pertuzumab-treated cells expressing both EGFR and ErbB2. By confocal microscopy analysis, more EGF was observed in endosomes on incubation with pertuzumab, and under similar conditions, immunoblotting experiments showed increased EGFR degradation on incubation with both EGF and pertuzumab. These results show that pertuzumab enhanced the endocytic down-regulation of EGFR by counteracting EGFR-ErbB2 heterodimerization. Our previous results showing that ErbB2 counteracts EGFR endocytosis can therefore be explained by tethering of EGFR to ErbB2 at the plasma membrane.
Hughes
Juliana Bentes
JB
Institute of Pathology, University of Oslo, Oslo, Norway.
Berger
Christian
C
Rødland
Marianne Skeie
MS
Hasmann
Max
M
Stang
Espen
E
Madshus
Inger Helene
IH
D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism*; D061067:Antibodies, Monoclonal, Humanized; D001011:Aorta / Q000166:cytology / Q000378:metabolism*; D002478:Cells, Cultured; D003432:Cross-Linking Reagents / Q000494:pharmacology; D015536:Down-Regulation; D004730:Endothelium, Vascular / Q000166:cytology / Q000378:metabolism*; D004815:Epidermal Growth Factor / Q000494:pharmacology; D066246:ErbB Receptors / Q000037:antagonists & inhibitors / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D007124:Immunoenzyme Techniques; D047468:Immunoprecipitation; D007457:Iodine Radioisotopes; D055503:Protein Multimerization; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013552:Swine
D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D003432:Cross-Linking Reagents; D004815:Epidermal Growth Factor; D066246:ErbB Receptors; D007457:Iodine Radioisotopes; D018719:Erb-b2 Receptor Tyrosine Kinases; D011994:Recombinant Proteins; C485206:pertuzumab