Integrated molecular and clinical analysis of AKT activation in metastatic melanoma

Michael A. Davies, Katherine Stemke-Hale, E. Lin, Carmen Tellez, Wanleng Deng, Yennu N. Gopal, Scott E. Woodman, Tiffany C. Calderone, Zhenlin Ju, Alexander J. Lazar, Victor G. Prieto, Kenneth Aldape, Gordon Mills, Jeffrey E. Gershenwald

Research output: Contribution to journalArticle

146 Citations (Scopus)

Abstract

Purpose: Activation of the phosphoinositide 3-kinase (PI3K)-AKT pathway has been implicated in melanoma based primarily on the prevalence of mutations in PTEN and NRAS. To improve our understanding of the regulation and clinical significance of the PI3K-AKT pathway in melanoma, we quantitatively measured the levels of phosphorylated AKT, its substrate GSK3α/β, and its negative regulator PTEN in clinical metastases. Results were compared with mutational status, clinical outcomes, and sites of metastasis. Experimental Design: DNA and protein were isolated from dissected frozen melanoma metastases (n = 96). Activating mutations of BRAF, NRAS, AKT, PIK3CA, and KIT were detected by mass spectroscopy genotyping. Phosphorylated AKT (Ser473 and Thr308), P-GSK3α/β, and PTEN protein expression were measured by reverse-phase protein array. A panel of human melanoma cells lines (n = 58) was analyzed for comparison. Results: BRAF-mutant tumors had higher levels of P-AKT-Ser473 (P = 0.01), P-AKTThr308 (P = 0.002), and P-GSK3α/β (P = 0.08) than NRAS-mutant tumors. Analysis of individual tumors showed that almost all tumors with elevated P-AKT had low PTEN levels; NRAS-mutant tumors had normal PTEN and lower P-AKT. Similar results were observed in melanoma cell lines. Stage III melanoma patients did not differ in overall survival based on activation status of the PI3K-AKT pathway. Brain metastases had significantly higher P-AKT and lower PTEN than lung or liver metastases. Conclusions: Quantitative interrogation of the PI3K-AKT pathway in melanoma reveals unexpected significant differences in AKT activation by NRAS mutation and PTEN loss, and hyperactivation of AKT in brain metastases. These findings have implications for the rational development of targeted therapy for this disease.

Original languageEnglish (US)
Pages (from-to)7538-7546
Number of pages9
JournalClinical Cancer Research
Volume15
Issue number24
DOIs
StatePublished - Dec 15 2009
Externally publishedYes

Fingerprint

Activation Analysis
Melanoma
1-Phosphatidylinositol 4-Kinase
Neoplasm Metastasis
Neoplasms
Mutation
PTEN Phosphohydrolase
Cell Line
Protein Array Analysis
Brain
Mass Spectrometry
Research Design
Lung
Survival
Liver
DNA

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Davies, M. A., Stemke-Hale, K., Lin, E., Tellez, C., Deng, W., Gopal, Y. N., ... Gershenwald, J. E. (2009). Integrated molecular and clinical analysis of AKT activation in metastatic melanoma. Clinical Cancer Research, 15(24), 7538-7546. https://doi.org/10.1158/1078-0432.CCR-09-1985

Integrated molecular and clinical analysis of AKT activation in metastatic melanoma. / Davies, Michael A.; Stemke-Hale, Katherine; Lin, E.; Tellez, Carmen; Deng, Wanleng; Gopal, Yennu N.; Woodman, Scott E.; Calderone, Tiffany C.; Ju, Zhenlin; Lazar, Alexander J.; Prieto, Victor G.; Aldape, Kenneth; Mills, Gordon; Gershenwald, Jeffrey E.

In: Clinical Cancer Research, Vol. 15, No. 24, 15.12.2009, p. 7538-7546.

Research output: Contribution to journalArticle

Davies, MA, Stemke-Hale, K, Lin, E, Tellez, C, Deng, W, Gopal, YN, Woodman, SE, Calderone, TC, Ju, Z, Lazar, AJ, Prieto, VG, Aldape, K, Mills, G & Gershenwald, JE 2009, 'Integrated molecular and clinical analysis of AKT activation in metastatic melanoma', Clinical Cancer Research, vol. 15, no. 24, pp. 7538-7546. https://doi.org/10.1158/1078-0432.CCR-09-1985
Davies, Michael A. ; Stemke-Hale, Katherine ; Lin, E. ; Tellez, Carmen ; Deng, Wanleng ; Gopal, Yennu N. ; Woodman, Scott E. ; Calderone, Tiffany C. ; Ju, Zhenlin ; Lazar, Alexander J. ; Prieto, Victor G. ; Aldape, Kenneth ; Mills, Gordon ; Gershenwald, Jeffrey E. / Integrated molecular and clinical analysis of AKT activation in metastatic melanoma. In: Clinical Cancer Research. 2009 ; Vol. 15, No. 24. pp. 7538-7546.
@article{0557bfee3ae4479aacf503e01979f8ec,
title = "Integrated molecular and clinical analysis of AKT activation in metastatic melanoma",
abstract = "Purpose: Activation of the phosphoinositide 3-kinase (PI3K)-AKT pathway has been implicated in melanoma based primarily on the prevalence of mutations in PTEN and NRAS. To improve our understanding of the regulation and clinical significance of the PI3K-AKT pathway in melanoma, we quantitatively measured the levels of phosphorylated AKT, its substrate GSK3α/β, and its negative regulator PTEN in clinical metastases. Results were compared with mutational status, clinical outcomes, and sites of metastasis. Experimental Design: DNA and protein were isolated from dissected frozen melanoma metastases (n = 96). Activating mutations of BRAF, NRAS, AKT, PIK3CA, and KIT were detected by mass spectroscopy genotyping. Phosphorylated AKT (Ser473 and Thr308), P-GSK3α/β, and PTEN protein expression were measured by reverse-phase protein array. A panel of human melanoma cells lines (n = 58) was analyzed for comparison. Results: BRAF-mutant tumors had higher levels of P-AKT-Ser473 (P = 0.01), P-AKTThr308 (P = 0.002), and P-GSK3α/β (P = 0.08) than NRAS-mutant tumors. Analysis of individual tumors showed that almost all tumors with elevated P-AKT had low PTEN levels; NRAS-mutant tumors had normal PTEN and lower P-AKT. Similar results were observed in melanoma cell lines. Stage III melanoma patients did not differ in overall survival based on activation status of the PI3K-AKT pathway. Brain metastases had significantly higher P-AKT and lower PTEN than lung or liver metastases. Conclusions: Quantitative interrogation of the PI3K-AKT pathway in melanoma reveals unexpected significant differences in AKT activation by NRAS mutation and PTEN loss, and hyperactivation of AKT in brain metastases. These findings have implications for the rational development of targeted therapy for this disease.",
author = "Davies, {Michael A.} and Katherine Stemke-Hale and E. Lin and Carmen Tellez and Wanleng Deng and Gopal, {Yennu N.} and Woodman, {Scott E.} and Calderone, {Tiffany C.} and Zhenlin Ju and Lazar, {Alexander J.} and Prieto, {Victor G.} and Kenneth Aldape and Gordon Mills and Gershenwald, {Jeffrey E.}",
year = "2009",
month = "12",
day = "15",
doi = "10.1158/1078-0432.CCR-09-1985",
language = "English (US)",
volume = "15",
pages = "7538--7546",
journal = "Clinical Cancer Research",
issn = "1078-0432",
publisher = "American Association for Cancer Research Inc.",
number = "24",

}

TY - JOUR

T1 - Integrated molecular and clinical analysis of AKT activation in metastatic melanoma

AU - Davies, Michael A.

AU - Stemke-Hale, Katherine

AU - Lin, E.

AU - Tellez, Carmen

AU - Deng, Wanleng

AU - Gopal, Yennu N.

AU - Woodman, Scott E.

AU - Calderone, Tiffany C.

AU - Ju, Zhenlin

AU - Lazar, Alexander J.

AU - Prieto, Victor G.

AU - Aldape, Kenneth

AU - Mills, Gordon

AU - Gershenwald, Jeffrey E.

PY - 2009/12/15

Y1 - 2009/12/15

N2 - Purpose: Activation of the phosphoinositide 3-kinase (PI3K)-AKT pathway has been implicated in melanoma based primarily on the prevalence of mutations in PTEN and NRAS. To improve our understanding of the regulation and clinical significance of the PI3K-AKT pathway in melanoma, we quantitatively measured the levels of phosphorylated AKT, its substrate GSK3α/β, and its negative regulator PTEN in clinical metastases. Results were compared with mutational status, clinical outcomes, and sites of metastasis. Experimental Design: DNA and protein were isolated from dissected frozen melanoma metastases (n = 96). Activating mutations of BRAF, NRAS, AKT, PIK3CA, and KIT were detected by mass spectroscopy genotyping. Phosphorylated AKT (Ser473 and Thr308), P-GSK3α/β, and PTEN protein expression were measured by reverse-phase protein array. A panel of human melanoma cells lines (n = 58) was analyzed for comparison. Results: BRAF-mutant tumors had higher levels of P-AKT-Ser473 (P = 0.01), P-AKTThr308 (P = 0.002), and P-GSK3α/β (P = 0.08) than NRAS-mutant tumors. Analysis of individual tumors showed that almost all tumors with elevated P-AKT had low PTEN levels; NRAS-mutant tumors had normal PTEN and lower P-AKT. Similar results were observed in melanoma cell lines. Stage III melanoma patients did not differ in overall survival based on activation status of the PI3K-AKT pathway. Brain metastases had significantly higher P-AKT and lower PTEN than lung or liver metastases. Conclusions: Quantitative interrogation of the PI3K-AKT pathway in melanoma reveals unexpected significant differences in AKT activation by NRAS mutation and PTEN loss, and hyperactivation of AKT in brain metastases. These findings have implications for the rational development of targeted therapy for this disease.

AB - Purpose: Activation of the phosphoinositide 3-kinase (PI3K)-AKT pathway has been implicated in melanoma based primarily on the prevalence of mutations in PTEN and NRAS. To improve our understanding of the regulation and clinical significance of the PI3K-AKT pathway in melanoma, we quantitatively measured the levels of phosphorylated AKT, its substrate GSK3α/β, and its negative regulator PTEN in clinical metastases. Results were compared with mutational status, clinical outcomes, and sites of metastasis. Experimental Design: DNA and protein were isolated from dissected frozen melanoma metastases (n = 96). Activating mutations of BRAF, NRAS, AKT, PIK3CA, and KIT were detected by mass spectroscopy genotyping. Phosphorylated AKT (Ser473 and Thr308), P-GSK3α/β, and PTEN protein expression were measured by reverse-phase protein array. A panel of human melanoma cells lines (n = 58) was analyzed for comparison. Results: BRAF-mutant tumors had higher levels of P-AKT-Ser473 (P = 0.01), P-AKTThr308 (P = 0.002), and P-GSK3α/β (P = 0.08) than NRAS-mutant tumors. Analysis of individual tumors showed that almost all tumors with elevated P-AKT had low PTEN levels; NRAS-mutant tumors had normal PTEN and lower P-AKT. Similar results were observed in melanoma cell lines. Stage III melanoma patients did not differ in overall survival based on activation status of the PI3K-AKT pathway. Brain metastases had significantly higher P-AKT and lower PTEN than lung or liver metastases. Conclusions: Quantitative interrogation of the PI3K-AKT pathway in melanoma reveals unexpected significant differences in AKT activation by NRAS mutation and PTEN loss, and hyperactivation of AKT in brain metastases. These findings have implications for the rational development of targeted therapy for this disease.

UR - http://www.scopus.com/inward/record.url?scp=73349128709&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=73349128709&partnerID=8YFLogxK

U2 - 10.1158/1078-0432.CCR-09-1985

DO - 10.1158/1078-0432.CCR-09-1985

M3 - Article

VL - 15

SP - 7538

EP - 7546

JO - Clinical Cancer Research

JF - Clinical Cancer Research

SN - 1078-0432

IS - 24

ER -