Phosphoproteomic mass spectrometry profiling links Src family kinases to escape from HER2 tyrosine kinase inhibition

B. N. Rexer, A. J.L. Ham, C. Rinehart, S. Hill, N. De Matos Granja-Ingram, A. M. González-Angulo, Gordon Mills, B. Dave, J. C. Chang, D. C. Liebler, C. L. Arteaga

Research output: Contribution to journalArticle

91 Citations (Scopus)

Abstract

Despite the initial effectiveness of the tyrosine kinase inhibitor lapatinib against HER2 gene-amplified breast cancers, most patients eventually relapse after treatment, implying that tumors acquire mechanisms of drug resistance. To discover these mechanisms, we generated six lapatinib-resistant HER2-overexpressing human breast cancer cell lines. In cells that grew in the presence of lapatinib, HER2 autophosphorylation was undetectable, whereas active phosphoinositide-3 kinase (PI3K)-Akt and mitogen-activated protein kinase (MAPK) were maintained. To identify networks maintaining these signaling pathways, we profiled the tyrosine phosphoproteome of sensitive and resistant cells using an immunoaffinity-enriched mass spectrometry method. We found increased phosphorylation of Src family kinases (SFKs) and putative Src substrates in several resistant cell lines. Treatment of these resistant cells with Src kinase inhibitors partially blocked PI3K-Akt signaling and restored lapatinib sensitivity. Further, SFK mRNA expression was upregulated in primary HER2 tumors treated with lapatinib. Finally, the combination of lapatinib and the Src inhibitor AZD0530 was more effective than lapatinib alone at inhibiting pAkt and growth of established HER2-positive BT-474 xenografts in athymic mice. These data suggest that increased Src kinase activity is a mechanism of lapatinib resistance and support the combination of HER2 antagonists with Src inhibitors early in the treatment of HER2 breast cancers in order to prevent or overcome resistance to HER2 inhibitors.

Original languageEnglish (US)
Pages (from-to)4163-4174
Number of pages12
JournalOncogene
Volume30
Issue number40
DOIs
StatePublished - Oct 6 2011
Externally publishedYes

Fingerprint

src-Family Kinases
Protein-Tyrosine Kinases
Mass Spectrometry
1-Phosphatidylinositol 4-Kinase
Breast Neoplasms
erbB-2 Genes
Cell Line
lapatinib
Mitogen-Activated Protein Kinases
Drug Resistance
Heterografts
Nude Mice
Tyrosine
Neoplasms
Therapeutics
Phosphorylation
Recurrence
Messenger RNA
Growth

Keywords

  • breast cancer
  • HER2
  • lapatinib
  • Src kinases
  • tyrosine phosphorylation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cancer Research

Cite this

Rexer, B. N., Ham, A. J. L., Rinehart, C., Hill, S., De Matos Granja-Ingram, N., González-Angulo, A. M., ... Arteaga, C. L. (2011). Phosphoproteomic mass spectrometry profiling links Src family kinases to escape from HER2 tyrosine kinase inhibition. Oncogene, 30(40), 4163-4174. https://doi.org/10.1038/onc.2011.130

Phosphoproteomic mass spectrometry profiling links Src family kinases to escape from HER2 tyrosine kinase inhibition. / Rexer, B. N.; Ham, A. J.L.; Rinehart, C.; Hill, S.; De Matos Granja-Ingram, N.; González-Angulo, A. M.; Mills, Gordon; Dave, B.; Chang, J. C.; Liebler, D. C.; Arteaga, C. L.

In: Oncogene, Vol. 30, No. 40, 06.10.2011, p. 4163-4174.

Research output: Contribution to journalArticle

Rexer, BN, Ham, AJL, Rinehart, C, Hill, S, De Matos Granja-Ingram, N, González-Angulo, AM, Mills, G, Dave, B, Chang, JC, Liebler, DC & Arteaga, CL 2011, 'Phosphoproteomic mass spectrometry profiling links Src family kinases to escape from HER2 tyrosine kinase inhibition', Oncogene, vol. 30, no. 40, pp. 4163-4174. https://doi.org/10.1038/onc.2011.130
Rexer BN, Ham AJL, Rinehart C, Hill S, De Matos Granja-Ingram N, González-Angulo AM et al. Phosphoproteomic mass spectrometry profiling links Src family kinases to escape from HER2 tyrosine kinase inhibition. Oncogene. 2011 Oct 6;30(40):4163-4174. https://doi.org/10.1038/onc.2011.130
Rexer, B. N. ; Ham, A. J.L. ; Rinehart, C. ; Hill, S. ; De Matos Granja-Ingram, N. ; González-Angulo, A. M. ; Mills, Gordon ; Dave, B. ; Chang, J. C. ; Liebler, D. C. ; Arteaga, C. L. / Phosphoproteomic mass spectrometry profiling links Src family kinases to escape from HER2 tyrosine kinase inhibition. In: Oncogene. 2011 ; Vol. 30, No. 40. pp. 4163-4174.
@article{1de266091aa043e79aed7d0afa01bcd2,
title = "Phosphoproteomic mass spectrometry profiling links Src family kinases to escape from HER2 tyrosine kinase inhibition",
abstract = "Despite the initial effectiveness of the tyrosine kinase inhibitor lapatinib against HER2 gene-amplified breast cancers, most patients eventually relapse after treatment, implying that tumors acquire mechanisms of drug resistance. To discover these mechanisms, we generated six lapatinib-resistant HER2-overexpressing human breast cancer cell lines. In cells that grew in the presence of lapatinib, HER2 autophosphorylation was undetectable, whereas active phosphoinositide-3 kinase (PI3K)-Akt and mitogen-activated protein kinase (MAPK) were maintained. To identify networks maintaining these signaling pathways, we profiled the tyrosine phosphoproteome of sensitive and resistant cells using an immunoaffinity-enriched mass spectrometry method. We found increased phosphorylation of Src family kinases (SFKs) and putative Src substrates in several resistant cell lines. Treatment of these resistant cells with Src kinase inhibitors partially blocked PI3K-Akt signaling and restored lapatinib sensitivity. Further, SFK mRNA expression was upregulated in primary HER2 tumors treated with lapatinib. Finally, the combination of lapatinib and the Src inhibitor AZD0530 was more effective than lapatinib alone at inhibiting pAkt and growth of established HER2-positive BT-474 xenografts in athymic mice. These data suggest that increased Src kinase activity is a mechanism of lapatinib resistance and support the combination of HER2 antagonists with Src inhibitors early in the treatment of HER2 breast cancers in order to prevent or overcome resistance to HER2 inhibitors.",
keywords = "breast cancer, HER2, lapatinib, Src kinases, tyrosine phosphorylation",
author = "Rexer, {B. N.} and Ham, {A. J.L.} and C. Rinehart and S. Hill and {De Matos Granja-Ingram}, N. and Gonz{\'a}lez-Angulo, {A. M.} and Gordon Mills and B. Dave and Chang, {J. C.} and Liebler, {D. C.} and Arteaga, {C. L.}",
year = "2011",
month = "10",
day = "6",
doi = "10.1038/onc.2011.130",
language = "English (US)",
volume = "30",
pages = "4163--4174",
journal = "Oncogene",
issn = "0950-9232",
publisher = "Nature Publishing Group",
number = "40",

}

TY - JOUR

T1 - Phosphoproteomic mass spectrometry profiling links Src family kinases to escape from HER2 tyrosine kinase inhibition

AU - Rexer, B. N.

AU - Ham, A. J.L.

AU - Rinehart, C.

AU - Hill, S.

AU - De Matos Granja-Ingram, N.

AU - González-Angulo, A. M.

AU - Mills, Gordon

AU - Dave, B.

AU - Chang, J. C.

AU - Liebler, D. C.

AU - Arteaga, C. L.

PY - 2011/10/6

Y1 - 2011/10/6

N2 - Despite the initial effectiveness of the tyrosine kinase inhibitor lapatinib against HER2 gene-amplified breast cancers, most patients eventually relapse after treatment, implying that tumors acquire mechanisms of drug resistance. To discover these mechanisms, we generated six lapatinib-resistant HER2-overexpressing human breast cancer cell lines. In cells that grew in the presence of lapatinib, HER2 autophosphorylation was undetectable, whereas active phosphoinositide-3 kinase (PI3K)-Akt and mitogen-activated protein kinase (MAPK) were maintained. To identify networks maintaining these signaling pathways, we profiled the tyrosine phosphoproteome of sensitive and resistant cells using an immunoaffinity-enriched mass spectrometry method. We found increased phosphorylation of Src family kinases (SFKs) and putative Src substrates in several resistant cell lines. Treatment of these resistant cells with Src kinase inhibitors partially blocked PI3K-Akt signaling and restored lapatinib sensitivity. Further, SFK mRNA expression was upregulated in primary HER2 tumors treated with lapatinib. Finally, the combination of lapatinib and the Src inhibitor AZD0530 was more effective than lapatinib alone at inhibiting pAkt and growth of established HER2-positive BT-474 xenografts in athymic mice. These data suggest that increased Src kinase activity is a mechanism of lapatinib resistance and support the combination of HER2 antagonists with Src inhibitors early in the treatment of HER2 breast cancers in order to prevent or overcome resistance to HER2 inhibitors.

AB - Despite the initial effectiveness of the tyrosine kinase inhibitor lapatinib against HER2 gene-amplified breast cancers, most patients eventually relapse after treatment, implying that tumors acquire mechanisms of drug resistance. To discover these mechanisms, we generated six lapatinib-resistant HER2-overexpressing human breast cancer cell lines. In cells that grew in the presence of lapatinib, HER2 autophosphorylation was undetectable, whereas active phosphoinositide-3 kinase (PI3K)-Akt and mitogen-activated protein kinase (MAPK) were maintained. To identify networks maintaining these signaling pathways, we profiled the tyrosine phosphoproteome of sensitive and resistant cells using an immunoaffinity-enriched mass spectrometry method. We found increased phosphorylation of Src family kinases (SFKs) and putative Src substrates in several resistant cell lines. Treatment of these resistant cells with Src kinase inhibitors partially blocked PI3K-Akt signaling and restored lapatinib sensitivity. Further, SFK mRNA expression was upregulated in primary HER2 tumors treated with lapatinib. Finally, the combination of lapatinib and the Src inhibitor AZD0530 was more effective than lapatinib alone at inhibiting pAkt and growth of established HER2-positive BT-474 xenografts in athymic mice. These data suggest that increased Src kinase activity is a mechanism of lapatinib resistance and support the combination of HER2 antagonists with Src inhibitors early in the treatment of HER2 breast cancers in order to prevent or overcome resistance to HER2 inhibitors.

KW - breast cancer

KW - HER2

KW - lapatinib

KW - Src kinases

KW - tyrosine phosphorylation

UR - http://www.scopus.com/inward/record.url?scp=85027940454&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85027940454&partnerID=8YFLogxK

U2 - 10.1038/onc.2011.130

DO - 10.1038/onc.2011.130

M3 - Article

VL - 30

SP - 4163

EP - 4174

JO - Oncogene

JF - Oncogene

SN - 0950-9232

IS - 40

ER -