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https://doi.org/10.1371/journal.pone.0009094
Title: | Characterization of notch1 antibodies that inhibit signaling of both normal and mutated notch1 receptors | Authors: | Aste-Am閦aga M. Zhang N. Lineberger J.E. Arnold B.A. Toner T.J. Gu M. Huang L. Vitelli S. Vo K.T. Haytko P. Zhao J.Z. Baleydier F. L'Heureux S. Wang H. Gordon W.R. Thoryk E. Andrawes M.B. Tiyanont K. Stegmaier K. Roti G. Ross K.N. Franlin L.L. Wang H. Wang F. Chastain M. Bett A.J. Audoly L.P. Aster J.C. Blacklow S.C. Huber H.E. |
Keywords: | epidermal growth factor gamma secretase inhibitor metalloproteinase monoclonal antibody monoclonal antibody Notch1 Notch1 receptor Notch2 receptor Notch3 receptor Notch4 receptor transcription factor HES 1 transcription factor HES 5 unclassified drug JAG2 protein, human ligand membrane protein monoclonal antibody Notch1 receptor signal peptide acute lymphoblastic leukemia angiogenesis article controlled study deltex1 gene gene gene expression human human cell ligand binding phage display point mutation protein domain reporter gene T lymphocyte tumor cell line animal antibody specificity binding competition binding site cell line cell proliferation cell strain 3T3 drug effect flow cytometry gene expression regulation genetics immunology metabolism mouse mutation pathology reverse transcription polymerase chain reaction signal transduction Metazoa 3T3 Cells Animals Antibodies, Monoclonal Antibody Specificity Binding Sites Binding, Competitive Cell Line Cell Line, Tumor Cell Proliferation Flow Cytometry Gene Expression Regulation, Neoplastic Humans Intercellular Signaling Peptides and Proteins Ligands Membrane Proteins Mice Mutation Precursor T-Cell Lymphoblastic Leukemia-Lymphoma Receptor, Notch1 Reverse Transcriptase Polymerase Chain Reaction Signal Transduction |
Issue Date: | 2010 | Citation: | Aste-Am閦aga M., Zhang N., Lineberger J.E., Arnold B.A., Toner T.J., Gu M., Huang L., Vitelli S., Vo K.T., Haytko P., Zhao J.Z., Baleydier F., L'Heureux S., Wang H., Gordon W.R., Thoryk E., Andrawes M.B., Tiyanont K., Stegmaier K., Roti G., Ross K.N., Franlin L.L., Wang H., Wang F., Chastain M., Bett A.J., Audoly L.P., Aster J.C., Blacklow S.C., Huber H.E. (2010). Characterization of notch1 antibodies that inhibit signaling of both normal and mutated notch1 receptors. PLoS ONE 5 (2) : e9094. ScholarBank@NUS Repository. https://doi.org/10.1371/journal.pone.0009094 | Rights: | Attribution 4.0 International | Abstract: | Background: Notch receptors normally play a key role in guiding a variety of cell fate decisions during development and differentiation of metazoan organisms. On the other hand, dysregulation of Notch1 signaling is associated with many different types of cancer as well as tumor angiogenesis, making Notch1 a potential therapeutic target. Principal Findings: Here we report the in vitro activities of inhibitory Notch1 monoclonal antibodies derived from cell-based and solid-phase screening of a phage display library. Two classes of antibodies were found, one directed against the EGF-repeat region that encompasses the ligand-binding domain (LBD), and the second directed against the activation switch of the receptor, the Notch negative regulatory region (NRR). The antibodies are selective for Notch1, inhibiting Jag2-dependent signaling by Notch1 but not by Notch 2 and 3 in reporter gene assays, with EC50 values as low as 5�nM and 0.13�09 nM for the LBD and NRR antibodies, respectively, and fail to recognize Notch4. While more potent, NRR antibodies are incomplete antagonists of Notch1 signaling. The antagonistic activity of LBD, but not NRR, antibodies is strongly dependent on the activating ligand. Both LBD and NRR antibodies bind to Notch1 on human tumor cell lines and inhibit the expression of sentinel Notch target genes, including HES1, HES5, and DTX1. NRR antibodies also strongly inhibit ligand-independent signaling in heterologous cells transiently expressing Notch1 receptors with diverse NRR "class I" point mutations, the most common type of mutation found in human T-cell acute lymphoblastic leukemia (T-ALL). In contrast, NRR antibodies failed to antagonize Notch1 receptors bearing rare "class II" or "class III" mutations, in which amino acid insertions generate a duplicated or constitutively sensitive metalloprotease cleavage site. Signaling in T-ALL cell lines bearing class I mutations is partially refractory to inhibitory antibodies as compared to cell-penetrating gamma-secretase inhibitors. Conclusions/Significance: Antibodies that compete with Notch1 ligand binding or that bind to the negative regulatory region can act as potent inhibitors of Notch1 signaling. These antibodies may have clinical utility for conditions in which inhibition of signaling by wild-type Notch1 is desired, but are likely to be of limited value for treatment of T-ALLs associated with aberrant Notch1 activation. � 2010 Aste-Amezaga et al. | Source Title: | PLoS ONE | URI: | https://scholarbank.nus.edu.sg/handle/10635/161817 | ISSN: | 19326203 | DOI: | 10.1371/journal.pone.0009094 | Rights: | Attribution 4.0 International |
Appears in Collections: | Elements Staff Publications |
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