title: High-throughput time-resolved FRET reveals Akt/PKB activation as a poor prognostic marker in breast cancer.
abstract: Dysregulation of the Akt/PKB pathway has been associated with poor prognosis in several human carcinomas. Current approaches to assess Akt activation rely on intensity-based methods, which are limited by the subjectivity of manual scoring and poor specificity. Here, we report the development of a novel assay using amplified, time-resolved Förster resonance energy transfer (FRET), which is highly specific and sensitive and can be adapted to any protein. Using this approach to analyze primary breast tissue microarrays, we quantified levels of activated pAkt at a spatial resolution that revealed molecular heterogeneity within tumors. High pAkt status assessed by amplified FRET correlated with worse disease-free survival. Our findings support the use of amplified FRET to determine pAkt status in cancer tissues as candidate biomarker for the identification of high-risk patients.
authors: Cell Biophysics Laboratory, Cancer Research UK, London Research Institute, London, United Kingdom.
authors: Selvaraju
authors: S
authors: Veeriah
authors: Cell Biophysics Laboratory, Cancer Research UK, London Research Institute, London, United Kingdom. Centre Emotion, Hôpital de la Pitié Salpetriere, Paris, France.
authors: Pierre
authors: P
authors: Leboucher
authors: Cell Biophysics Laboratory, Cancer Research UK, London Research Institute, London, United Kingdom. Protein Phosphorylation Laboratory, Cancer Research UK, London Research Institute, London, United Kingdom.
authors: Julien
authors: J
authors: de Naurois
authors: Cell Biophysics Laboratory, Cancer Research UK, London Research Institute, London, United Kingdom.
authors: Nirmal
authors: N
authors: Jethwa
authors: Experimental Histopathology Laboratory, Cancer Research UK, London Research Institute, London, United Kingdom.
authors: Emma
authors: E
authors: Nye
authors: Experimental Histopathology Laboratory, Cancer Research UK, London Research Institute, London, United Kingdom.
authors: Tamara
authors: T
authors: Bunting
authors: Experimental Histopathology Laboratory, Cancer Research UK, London Research Institute, London, United Kingdom.
authors: Richard
authors: R
authors: Stone
authors: Experimental Histopathology Laboratory, Cancer Research UK, London Research Institute, London, United Kingdom.
authors: Gordon
authors: G
authors: Stamp
authors: Cell Biophysics Laboratory, Cancer Research UK, London Research Institute, London, United Kingdom.
authors: Véronique
authors: V
authors: Calleja
authors: Stanford University School of Medicine, Stanford, California.
authors: Stefanie S
authors: SS
authors: Jeffrey
authors: Protein Phosphorylation Laboratory, Cancer Research UK, London Research Institute, London, United Kingdom. Cancer Division King's College London, London, United Kingdom.
authors: Peter J
authors: PJ
authors: Parker
authors: Cell Biophysics Laboratory, Cancer Research UK, London Research Institute, London, United Kingdom. Cell Biophysics Laboratory, Unidad de Biofísica (CSIC-UPV/EHU), Sarriena s/n, Leioa, Spain. Ikerbasque, Basque Foundation for Science, University of the Basque Country, Sarriena s/n, Leioa, Spain. banafshe.larijani@ikerbasque.org.
authors: Banafshé
authors: B
authors: Larijani
mesh_terms: D014408:Biomarkers, Tumor / Q000032:analysis* / Q000378:metabolism; D001943:Breast Neoplasms / Q000201:enzymology* / Q000473:pathology; D045744:Cell Line, Tumor; D018572:Disease-Free Survival; D005260:Female; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D011379:Prognosis; D051057:Proto-Oncogene Proteins c-akt / Q000032:analysis* / Q000378:metabolism
chemical_list: D014408:Biomarkers, Tumor; D051057:Proto-Oncogene Proteins c-akt