Time-resolved fluorescence resonance energy transfer (TR-FRET) to analyze the disruption of EGFR/HER2 dimers: a new method to evaluate the efficiency of targeted therapy using monoclonal antibodies.: TR-FRET to quantify HER dimers and evaluate mAb efficiency
| Autor: | Gaborit, Nadège, Larbouret, Christel, Vallaghe, Julie, Peyrusson, Frédéric, Bascoul-Mollevi, Caroline, Crapez, Evelyne, Azria, David, Chardes, Thierry, Poul, Marie-Alix, Mathis, Gérard, Bazin, Hervé, Pèlegrin, André |
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| Přispěvatelé: | Le Ster, Yves, Institut de recherche en cancérologie de Montpellier (IRCM - U896 Inserm - UM1), Université Montpellier 1 (UM1)-CRLCC Val d'Aurelle - Paul Lamarque-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM), CRLC Val d'Aurelle-Paul Lamarque, CRLCC Val d'Aurelle - Paul Lamarque, Cisbio, Research Department, CIS BIOINTERNATIONAL |
| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
MESH: Cell Line
Tumor [SDV.IMM] Life Sciences [q-bio]/Immunology MESH: Fluorescence Resonance Energy Transfer [SDV.CAN]Life Sciences [q-bio]/Cancer [SDV.BC]Life Sciences [q-bio]/Cellular Biology MESH: Receptors Fibroblast Growth Factor [SDV.CAN] Life Sciences [q-bio]/Cancer MESH: Cell Proliferation [SDV.BBM] Life Sciences [q-bio]/Biochemistry Molecular Biology MESH: Protein Kinase Inhibitors MESH: Animals MESH: Neoplasms [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology skin and connective tissue diseases neoplasms [SDV.BC] Life Sciences [q-bio]/Cellular Biology MESH: Mice MESH: Humans MESH: Phosphorylation MESH: Protein Multimerization MESH: Drug Screening Assays Antitumor [SDV.SP]Life Sciences [q-bio]/Pharmaceutical sciences [SDV.SP] Life Sciences [q-bio]/Pharmaceutical sciences MESH: Quinazolines MESH: Antibodies Neoplasm MESH: Antibodies Monoclonal Murine-Derived MESH: Receptor erbB-2 MESH: Antineoplastic Agents [SDV.IMM]Life Sciences [q-bio]/Immunology MESH: NIH 3T3 Cells |
| Zdroj: | Journal of Biological Chemistry Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2011, 286 (13), pp.11337-45. ⟨10.1074/jbc.M111.223503⟩ |
| ISSN: | 0021-9258 1083-351X |
| DOI: | 10.1074/jbc.M111.223503 |
| Popis: | International audience; In oncology, simultaneous inhibition of epidermal growth factor receptor (EGFR) and HER2 by monoclonal antibodies (mAbs) is an efficient therapeutic strategy but the underlying mechanisms are not fully understood. Here, we describe a time-resolved fluorescence resonance energy transfer (TR-FRET) method to quantify EGFR/HER2 heterodimers on cell surface to shed some light on the mechanism of such therapies. First, we tested this antibody-based TR-FRET assay in NIH/3T3 cell lines that express EGFR and/or HER2 and in various tumor cell lines. Then, we used the antibody-based TR-FRET assay to evaluate in vitro the effect of different targeted therapies on EGFR/HER2 heterodimers in the ovarian carcinoma cell line SKOV-3. A simultaneous incubation with Cetuximab (anti-EGFR) and Trastuzumab (anti-HER2) disturbed EGFR/HER2 heterodimers resulting in a 72% reduction. Cetuximab, Trastuzumab or Pertuzumab (anti-HER2) alone induced a 48, 44, or 24% reduction, respectively. In contrast, the tyrosine kinase inhibitors Erlotinib and Lapatinib had very little effect on EGFR/HER2 dimers concentration. In vivo, the combination of Cetuximab and Trastuzumab showed a better therapeutic effect (median survival and percentage of tumor-free mice) than the single mAbs. These results suggest a correlation between the extent of the mAb-induced EGFR/HER2 heterodimer reduction and the efficacy of such mAbs in targeted therapies. In conclusion, quantifying EGFR/HER2 heterodimers using our antibody-based TR-FRET assay may represent a useful method to predict the efficacy and explain the mechanisms of action of therapeutic mAbs, in addition to other commonly used techniques that focus on antibody-dependent cellular cytotoxicity, phosphorylation, and cell proliferation. |
| Databáze: | OpenAIRE |
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