Neutralization of the γ-secretase activity by monoclonal antibody against extracellular domain of nicastrin

  • Oncogene. 2012 Feb 9;31(6):787-798. doi: 10.1038/onc.2011.265.
I Hayashi  #  1 ,  S Takatori  #  1 ,  Y Urano  2 ,  Y Miyake  3 ,  J Takagi  4 ,  M Sakata-Yanagimoto  3 ,  H Iwanari  2  5 ,  S Osawa  1 ,  Y Morohashi  1 ,  T Li  6 ,  P C Wong  6 ,  S Chiba  3 ,  T Kodama  2 ,  T Hamakubo  2 ,  T Tomita  1  7 ,  T Iwatsubo  1  7  8
Affiliations
  • 1. Department of Neuropathology and Neuroscience, Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo, Japan.
  • 2. Department of Systems Biology and Medicine, Research Center for Advanced Science and Technology, The University of Tokyo, Tokyo, Japan.
  • 3. Department of Clinical and Experimental Hematology, Graduate School of Comprehensive Human Sciences, University of Tsukuba, Tsukuba, Japan.
  • 4. Laboratory of Protein Synthesis and Expression, Institute for Protein Research, Osaka University, Osaka, Japan.
  • 5. Perseus Proteomics Inc., Tokyo, Japan.
  • 6. Department of Pathology, The Johns Hopkins University, School of Medicine, Baltimore, MD, USA.
  • 7. Core Research for Evolutional Science and Technology, Japan Science and Technology Agency, Saitama, Japan.
  • 8. Department of Neuropathology, Graduate School of Medicine, The University of Tokyo, Tokyo, Japan.
  • # Contributed equally.
PMID: 21725355 DOI: 10.1038/onc.2011.265
Abstract

Several lines of evidence suggest that aberrant Notch signaling contributes to the development of several types of Cancer. Activation of Notch receptor is executed through intramembrane proteolysis by γ-secretase, which is a multimeric membrane-embedded protease comprised of presenilin, nicastrin (NCT), anterior pharynx defective 1 and PEN-2. In this study, we report the neutralization of the γ-secretase activity by a novel monoclonal antibody A5226A against the extracellular domain of NCT, generated by using a recombinant budded baculovirus as an immunogen. This antibody recognized fully glycosylated mature NCT in the active γ-secretase complex on the cell surface, and inhibited the γ-secretase activity by competing with the substrate binding in vitro. Moreover, A5226A abolished the γ-secretase activity-dependent growth of Cancer cells in a xenograft model. Our data provide compelling evidence that NCT is a molecular target for the mechanism-based inhibition of γ-secretase, and that targeting NCT might be a novel therapeutic strategy against Cancer caused by aberrant γ-secretase activity and Notch signaling.

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