Development of parallel reaction monitoring (PRM)-based quantitative proteomics applied to HER2-Positive breast cancer
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Mathilde Guerin1,2, Anthony Gonçalves1,2,6, Yves Toiron2, Emilie Baudelet2, Matthieu Pophillat2, Samuel Granjeaud2, Patrick Fourquet2, William Jacot3, Carole Tarpin1, Renaud Sabatier1,6, Emilie Agavnian4, Pascal Finetti6, José Adelaide6, Daniel Birnbaum6, Christophe Ginestier5, Emmanuelle Charafe-Jauffret4,5, Patrice Viens1,6, François Bertucci1,6, Jean-Paul Borg2 and Luc Camoin2
1Institut Paoli-Calmettes, Department of Medical Oncology, Marseille, France
2Aix-Marseille University, Inserm, CNRS, Institut Paoli-Calmettes, CRCM, Marseille Proteomics, Marseille, France
3IRCM, INSERM, Institut Régional du Cancer, Department of Medical Oncology, Montpellier, France
4Institut Paoli-Calmettes, Department of Anatomo-pathology, Marseille, France
5Aix-Marseille University, Inserm, CNRS, Institut Paoli-Calmettes, CRCM, Epithelial Stem Cells and Cancer Team, Marseille, France
6Aix-Marseille University, Inserm, CNRS, Institut Paoli-Calmettes, CRCM, Predictive Oncology Team, Marseille, France
Anthony Gonçalves, email: email@example.com
Luc Camoin, email: firstname.lastname@example.org
Keywords: proteomics; parallel reaction monitoring; mass spectrometry; breast cancer; HER2-positive
Received: March 19, 2018 Accepted: August 04, 2018 Published: September 18, 2018
Introduction: treatments targeting the Human Epidermal Growth Factor Receptor 2 (HER2/ERBB2) have improved the natural history of HER2-positive breast cancer. However, except HER2 protein expression and gene amplification, there is no predictive biomarker to guide the HER2-targeted therapies. We developed Parallel reaction monitoring (PRM) a powerful approach, to quantify and evaluate key proteins involved in the HER2 pathway and/or anti-HER2 treatment sensitivity.
Results: in BCLs, PRM measurements correlated with western blot immunocytochemistry and transcriptomic data. At baseline, higher expression of HER2, EGFR, PTEN and HER3 but lower expression of phospho-HER2 correlated with trastuzumab sensitivity. Under trastuzumab, PRM demonstrated a decrease in HER2 and an increase in phospho-HER2, which correlated with drug sensitivity. The opposite was observed under lapatinib. HER2 quantification was also correlated with immunohistochemistry in PDXs and clinical breast cancer samples.
Discussion: in conclusion, PRM-based assay, developed to quantify proteins of the HER2 pathway in breast cancer samples revealed a large magnitude of expression, which may have relevance in terms of treatment sensitivity.
Materials and Methods: we first evaluated PRM in term of sensitivity, linearity and reproducibility. PRM was then applied to breast cancer cell lines (BCLs) including BCLs exposed to anti-HER2 agents, patient-derived xenografts (PDXs) and frozen breast cancer samples.
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