Time resolved amplified FRET identifies protein kinase B activation state as a marker for poor prognosis in clear cell renal cell carcinoma

James Miles, Christopher Applebee, Pierre Leboucher, Sonia Lopez-Fernandez, Dae-Jin Lee, Rosa Guarch, Stephen Ward, Peter Parker, Jose Lopez, Banafshe Larijani

Research output: Contribution to journalArticle

Abstract

Purpose: Clear cell Renal Cell Carcinomas (ccRCC), the largest group of renal tumours, are resistant to classical therapies. The determination of the functional state of actionable biomarkers for the assessment of these adenocarcinomas is essential. The dysregulation of the oncoprotein, PKB/Akt has been linked with poor prognoses in human cancers. Material & Methods: We analysed the status of the PKB/Akt pathway in a representative tumour tissue microarray obtained from the primary tumours and their metastases in 60 ccRCC with long term follow up. We sought to define the evolution of this pathway from the primary tumour to the metastatic event and to know the impact of its functional state in tumour aggressiveness and patient survival. Two-site time resolved amplified FRET (A-FRET) was utilised for assessing the activation state of PKB/Akt and this was compared to conventional immunohistochemistry measurements. Results: Activation state of PKB/Akt in primary tumours defined by A-FRET correlated with poorer overall survival (hazard ratio 0.228; p=0.002). Whereas, increased protein expression of phosphoPKB/Akt, identified using classical immunohistochemistry, yielded no significant difference (hazard ratio 1.390; p=0.548). Conclusions: Quantitative determination of PKB/Akt activation in ccRCC primary tumours alongside other diagnostics tools could prove key in taking oncologists closer to an efficient personalised therapy in ccRCC patients. General Significance The quantitative imaging technology based on Amplified-FRET can rapidly analyse protein activation states and molecular interactions. It could be used for prognosis and assess drug function during the early cycles of chemotherapy. It enables evaluation of clinical efficiency of personalised cancer treatment.
Original languageEnglish
Pages (from-to)97-102
Number of pages14
JournalBBA Clinical
Volume8
Early online date13 Oct 2017
DOIs
Publication statusPublished - 1 Dec 2017

Fingerprint

Proto-Oncogene Proteins c-akt
Renal Cell Carcinoma
Neoplasms
Immunohistochemistry
Activation Analysis
Survival
Oncogene Proteins
Proteins
Adenocarcinoma
Therapeutics
Biomarkers
Neoplasm Metastasis
Technology
Kidney
Drug Therapy

Cite this

Time resolved amplified FRET identifies protein kinase B activation state as a marker for poor prognosis in clear cell renal cell carcinoma. / Miles, James; Applebee, Christopher ; Leboucher, Pierre; Lopez-Fernandez, Sonia; Lee, Dae-Jin; Guarch, Rosa; Ward, Stephen; Parker, Peter ; Lopez, Jose; Larijani, Banafshe.

In: BBA Clinical, Vol. 8, 01.12.2017, p. 97-102.

Research output: Contribution to journalArticle

Miles, James ; Applebee, Christopher ; Leboucher, Pierre ; Lopez-Fernandez, Sonia ; Lee, Dae-Jin ; Guarch, Rosa ; Ward, Stephen ; Parker, Peter ; Lopez, Jose ; Larijani, Banafshe. / Time resolved amplified FRET identifies protein kinase B activation state as a marker for poor prognosis in clear cell renal cell carcinoma. In: BBA Clinical. 2017 ; Vol. 8. pp. 97-102.
@article{5029b21478314c3fb08ce846e6738be6,
title = "Time resolved amplified FRET identifies protein kinase B activation state as a marker for poor prognosis in clear cell renal cell carcinoma",
abstract = "Purpose: Clear cell Renal Cell Carcinomas (ccRCC), the largest group of renal tumours, are resistant to classical therapies. The determination of the functional state of actionable biomarkers for the assessment of these adenocarcinomas is essential. The dysregulation of the oncoprotein, PKB/Akt has been linked with poor prognoses in human cancers. Material & Methods: We analysed the status of the PKB/Akt pathway in a representative tumour tissue microarray obtained from the primary tumours and their metastases in 60 ccRCC with long term follow up. We sought to define the evolution of this pathway from the primary tumour to the metastatic event and to know the impact of its functional state in tumour aggressiveness and patient survival. Two-site time resolved amplified FRET (A-FRET) was utilised for assessing the activation state of PKB/Akt and this was compared to conventional immunohistochemistry measurements. Results: Activation state of PKB/Akt in primary tumours defined by A-FRET correlated with poorer overall survival (hazard ratio 0.228; p=0.002). Whereas, increased protein expression of phosphoPKB/Akt, identified using classical immunohistochemistry, yielded no significant difference (hazard ratio 1.390; p=0.548). Conclusions: Quantitative determination of PKB/Akt activation in ccRCC primary tumours alongside other diagnostics tools could prove key in taking oncologists closer to an efficient personalised therapy in ccRCC patients. General Significance The quantitative imaging technology based on Amplified-FRET can rapidly analyse protein activation states and molecular interactions. It could be used for prognosis and assess drug function during the early cycles of chemotherapy. It enables evaluation of clinical efficiency of personalised cancer treatment.",
author = "James Miles and Christopher Applebee and Pierre Leboucher and Sonia Lopez-Fernandez and Dae-Jin Lee and Rosa Guarch and Stephen Ward and Peter Parker and Jose Lopez and Banafshe Larijani",
year = "2017",
month = "12",
day = "1",
doi = "10.1016/j.bbacli.2017.10.002",
language = "English",
volume = "8",
pages = "97--102",
journal = "BBA Clinical",
issn = "2214-6474",
publisher = "Elsevier",

}

TY - JOUR

T1 - Time resolved amplified FRET identifies protein kinase B activation state as a marker for poor prognosis in clear cell renal cell carcinoma

AU - Miles, James

AU - Applebee, Christopher

AU - Leboucher, Pierre

AU - Lopez-Fernandez, Sonia

AU - Lee, Dae-Jin

AU - Guarch, Rosa

AU - Ward, Stephen

AU - Parker, Peter

AU - Lopez, Jose

AU - Larijani, Banafshe

PY - 2017/12/1

Y1 - 2017/12/1

N2 - Purpose: Clear cell Renal Cell Carcinomas (ccRCC), the largest group of renal tumours, are resistant to classical therapies. The determination of the functional state of actionable biomarkers for the assessment of these adenocarcinomas is essential. The dysregulation of the oncoprotein, PKB/Akt has been linked with poor prognoses in human cancers. Material & Methods: We analysed the status of the PKB/Akt pathway in a representative tumour tissue microarray obtained from the primary tumours and their metastases in 60 ccRCC with long term follow up. We sought to define the evolution of this pathway from the primary tumour to the metastatic event and to know the impact of its functional state in tumour aggressiveness and patient survival. Two-site time resolved amplified FRET (A-FRET) was utilised for assessing the activation state of PKB/Akt and this was compared to conventional immunohistochemistry measurements. Results: Activation state of PKB/Akt in primary tumours defined by A-FRET correlated with poorer overall survival (hazard ratio 0.228; p=0.002). Whereas, increased protein expression of phosphoPKB/Akt, identified using classical immunohistochemistry, yielded no significant difference (hazard ratio 1.390; p=0.548). Conclusions: Quantitative determination of PKB/Akt activation in ccRCC primary tumours alongside other diagnostics tools could prove key in taking oncologists closer to an efficient personalised therapy in ccRCC patients. General Significance The quantitative imaging technology based on Amplified-FRET can rapidly analyse protein activation states and molecular interactions. It could be used for prognosis and assess drug function during the early cycles of chemotherapy. It enables evaluation of clinical efficiency of personalised cancer treatment.

AB - Purpose: Clear cell Renal Cell Carcinomas (ccRCC), the largest group of renal tumours, are resistant to classical therapies. The determination of the functional state of actionable biomarkers for the assessment of these adenocarcinomas is essential. The dysregulation of the oncoprotein, PKB/Akt has been linked with poor prognoses in human cancers. Material & Methods: We analysed the status of the PKB/Akt pathway in a representative tumour tissue microarray obtained from the primary tumours and their metastases in 60 ccRCC with long term follow up. We sought to define the evolution of this pathway from the primary tumour to the metastatic event and to know the impact of its functional state in tumour aggressiveness and patient survival. Two-site time resolved amplified FRET (A-FRET) was utilised for assessing the activation state of PKB/Akt and this was compared to conventional immunohistochemistry measurements. Results: Activation state of PKB/Akt in primary tumours defined by A-FRET correlated with poorer overall survival (hazard ratio 0.228; p=0.002). Whereas, increased protein expression of phosphoPKB/Akt, identified using classical immunohistochemistry, yielded no significant difference (hazard ratio 1.390; p=0.548). Conclusions: Quantitative determination of PKB/Akt activation in ccRCC primary tumours alongside other diagnostics tools could prove key in taking oncologists closer to an efficient personalised therapy in ccRCC patients. General Significance The quantitative imaging technology based on Amplified-FRET can rapidly analyse protein activation states and molecular interactions. It could be used for prognosis and assess drug function during the early cycles of chemotherapy. It enables evaluation of clinical efficiency of personalised cancer treatment.

U2 - 10.1016/j.bbacli.2017.10.002

DO - 10.1016/j.bbacli.2017.10.002

M3 - Article

VL - 8

SP - 97

EP - 102

JO - BBA Clinical

JF - BBA Clinical

SN - 2214-6474

ER -