Combination antibody treatment down-regulates epidermal growth factor receptor by inhibiting endosomal recycling

Edited* by John Kuriyan, University of California, Berkeley, CA, and approved June 4, 2010 (received for review November 20, 2009)
July 7, 2010
107 (30) 13252-13257

Abstract

Due to its common dysregulation in epithelial-based cancers and extensive characterization of its role in tumor growth, epidermal growth factor receptor (EGFR) is a highly validated target for anticancer therapies. There has been particular interest in the development of monoclonal antibodies (mAbs) targeting EGFR, resulting in two approved mAb-based drugs and several others in clinical trials. It has recently been reported that treatment with combinations of noncompetitive mAbs can induce receptor clustering, leading to synergistic receptor down-regulation. We elucidate three key aspects of this phenomenon. First, we show that highly potent combinations consisting of two noncompetitive mAbs that target EGFR domain 3 reduce surface receptor levels by up to 80% with a halftime of 0.5–5 h in both normal and transformed human cell lines to an extent inversely proportional to receptor density. Second, we find the mechanism underlying down-regulation to be consistent with recycling inhibition. Third, in contrast to the agonism associated with ligand-induced down-regulation, we demonstrate that mAb-induced down-regulation does not activate EGFR or its downstream effectors and it leads to synergistic reduction in migration and proliferation of cells that secrete autocrine ligand. These new insights will aid in ongoing rational design of EGFR-targeted antibody therapeutics.

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Acknowledgments.

The authors gratefully acknowledge Eliza Vasile and Michele Griffin of the Koch Center microscopy and flow cytometry facilities for technical assistance and Dr. Benjamin Hackel for valuable discussion and input. This work was funded by National Institutes of Health Grant CA96504 and a National Defense Science and Engineering Graduate Fellowship to J.B.S.

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Information & Authors

Information

Published in

Go to Proceedings of the National Academy of Sciences
Proceedings of the National Academy of Sciences
Vol. 107 | No. 30
July 27, 2010
PubMed: 20616078

Classifications

Submission history

Published online: July 7, 2010
Published in issue: July 27, 2010

Keywords

  1. ErbB
  2. monoclonal antibody
  3. tyrosine kinase
  4. trafficking

Acknowledgments

The authors gratefully acknowledge Eliza Vasile and Michele Griffin of the Koch Center microscopy and flow cytometry facilities for technical assistance and Dr. Benjamin Hackel for valuable discussion and input. This work was funded by National Institutes of Health Grant CA96504 and a National Defense Science and Engineering Graduate Fellowship to J.B.S.

Notes

*This Direct Submission article had a prearranged editor.

Authors

Affiliations

Jamie B. Spangler
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139;
Jason R. Neil
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139;
Sivan Abramovitch
Department of Biological Regulation, The Weizmann Institute of Science, Rehovot 76100, Israel; and
Yosef Yarden
Department of Biological Regulation, The Weizmann Institute of Science, Rehovot 76100, Israel; and
Forest M. White
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139;
Douglas A. Lauffenburger
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139;
K. Dane Wittrup1 [email protected]
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139;
Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139

Notes

1
To whom correspondence should be addressed at: Massachusetts Institute of Technology, 77 Massachusetts Avenue, E19-551, Cambridge, MA 02139. E-mail: [email protected].
Author contributions: J.B.S., D.A.L., and K.D.W. designed research; J.B.S. performed research; J.R.N., S.A., Y.Y., and F.M.W. contributed new reagents/analytic tools; J.B.S., F.M.W., D.A.L., and K.D.W. analyzed data; and J.B.S., F.M.W., D.A.L., and K.D.W. wrote the paper.

Competing Interests

The authors declare no conflict of interest.

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    Combination antibody treatment down-regulates epidermal growth factor receptor by inhibiting endosomal recycling
    Proceedings of the National Academy of Sciences
    • Vol. 107
    • No. 30
    • pp. 13191-13556

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