High-throughput time-resolved FRET reveals Akt/PKB activation as a poor prognostic marker in breast cancer

Selvaraju Veeriah, Pierre Leboucher, Julien de Naurois, Nirmal Jethwa, Emma Nye, Tamara Bunting, Richard Stone, Gordon Stamp, Véronique Calleja, Stefanie S Jeffrey, Peter J Parker, Banafshé Larijani

Research output: Contribution to journalArticlepeer-review

21 Citations (SciVal)


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.

Original languageEnglish
Pages (from-to)4983-4995
Number of pages13
JournalCancer Research
Issue number18
Publication statusPublished - 15 Sept 2014


  • Biomarkers, Tumor/analysis
  • Breast Neoplasms/enzymology
  • Cell Line, Tumor
  • Disease-Free Survival
  • Female
  • Fluorescence Resonance Energy Transfer/methods
  • High-Throughput Screening Assays/methods
  • Humans
  • Prognosis
  • Proto-Oncogene Proteins c-akt/analysis


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