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

TY - JOUR

T1 - High-throughput time-resolved FRET reveals Akt/PKB activation as a poor prognostic marker in breast cancer

AU - Veeriah, Selvaraju

AU - Leboucher, Pierre

AU - de Naurois, Julien

AU - Jethwa, Nirmal

AU - Nye, Emma

AU - Bunting, Tamara

AU - Stone, Richard

AU - Stamp, Gordon

AU - Calleja, Veronique

AU - Jeffrey, Stefanie S

AU - Parker, Peter J.

AU - Larijani, Banafshe

PY - 2014/9/15

Y1 - 2014/9/15

N2 - 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, timeresolved Forster 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.

AB - 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, timeresolved Forster 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.

U2 - 10.1158/0008-5472.CAN-13-3382

DO - 10.1158/0008-5472.CAN-13-3382

M3 - Article

SN - 0008-5472

VL - 74

SP - 4983

EP - 4995

JO - Cancer Research

JF - Cancer Research

IS - 18

ER -