title: Annotation of human cancers with EGFR signaling-associated protein complexes using proximity ligation assays.
abstract: Strategies to measure functional signaling-associated protein complexes have the potential to augment current molecular biomarker assays, such as genotyping and expression profiling, used to annotate diseases. Aberrant activation of epidermal growth factor receptor (EGFR) signaling contributes to diverse cancers. We used a proximity ligation assay (PLA) to detect EGFR in a complex with growth factor receptor-bound protein 2 (GRB2), the major signaling adaptor for EGFR. We used multiple lung cancer cell lines to develop and characterize EGFR:GRB2 PLA and correlated this assay with established biochemical measures of EGFR signaling. In a panel of patient-derived xenografts in mice, the intensity of EGFR:GRB2 PLA correlated with the reduction in tumor size in response to the EGFR inhibitor cetuximab. In tumor biopsies from three cohorts of lung cancer patients, positive EGFR:GRB2 PLA was observed in patients with and without EGFR mutations, and the intensity of EGFR:GRB2 PLA was predictive of overall survival in an EGFR inhibitor-treated cohort. Thus, we established the feasibility of using PLA to measure EGFR signaling-associated protein complexes in patient-based materials, suggesting the potential for similar assays for a broader array of receptor tyrosine kinases and other key signaling molecules.
authors.affiliation: Department of Thoracic Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA.
authors.forename: Matthew A
authors.initials: MA
authors.lastname: Smith
authors.affiliation: Graduate Medical Education, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA.
authors.forename: Richard
authors.initials: R
authors.lastname: Hall
authors.affiliation: Department of Biostatistics, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA.
authors.forename: Kate
authors.initials: K
authors.lastname: Fisher
authors.affiliation: Graduate Medical Education, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA.
authors.forename: Scott M
authors.initials: SM
authors.lastname: Haake
authors.affiliation: Department of Pathology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA.
authors.forename: Farah
authors.initials: F
authors.lastname: Khalil
authors.affiliation: Department of Cancer Epidemiology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA.
authors.forename: Matthew B
authors.initials: MB
authors.lastname: Schabath
authors.affiliation: Oncotest GmbH, Am Flughafen 12-14, 79108 Freiburg, Germany.
authors.forename: Vincent
authors.initials: V
authors.lastname: Vuaroqueaux
authors.affiliation: Oncotest GmbH, Am Flughafen 12-14, 79108 Freiburg, Germany.
authors.forename: Heinz-Herbert
authors.initials: HH
authors.lastname: Fiebig
authors.affiliation: Department of Pathology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA.
authors.forename: Soner
authors.initials: S
authors.lastname: Altiok
authors.affiliation: Department of Biostatistics, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA.
authors.forename: Yian Ann
authors.initials: YA
authors.lastname: Chen
authors.affiliation: Department of Thoracic Oncology, H. Lee Moffitt Cancer Center & Research Institute, Tampa, FL 33612, USA. eric.haura@moffitt.org.
authors.forename: Eric B
authors.initials: EB
authors.lastname: Haura
mesh_terms: D000818:Animals; D061067:Antibodies, Monoclonal, Humanized / Q000494:pharmacology; D014408:Biomarkers, Tumor / Q000378:metabolism*; D045744:Cell Line, Tumor; D000068818:Cetuximab; D066246:ErbB Receptors / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D051380:GRB2 Adaptor Protein / Q000378:metabolism; D064593:Heterografts / Q000187:drug effects / Q000473:pathology; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D008175:Lung Neoplasms / Q000175:diagnosis* / Q000378:metabolism; D051379:Mice; D046912:Multiprotein Complexes / Q000378:metabolism* / Q000502:physiology; D025941:Protein Interaction Mapping / Q000379:methods*; D015398:Signal Transduction / Q000502:physiology*
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D014408:Biomarkers, Tumor; D051380:GRB2 Adaptor Protein; D046912:Multiprotein Complexes; D066246:ErbB Receptors; D000068818:Cetuximab