Synergistic activation upon MET and ALK coamplification sustains targeted therapy in sarcomatoid Carcinoma, a deadly subtype of lung cancer

Giuseppe Pelosi, Patrizia Gasparini, Davide Conte, Alessandra Fabbri, Federica Perrone, Elena Tamborini, Serenella Pupa, Valentina Ciravolo, Roberto Caserini, Giulio Rossi, Alberto Cavazza, M. Papotti, Yukio Nakatani, Patrick Maisonneuve, Ugo Pastorino, Gabriella Sozzi

Research output: Contribution to journalArticle

Abstract

Introduction: Genetic alterations suitable for targeted therapy are poorly known issues in pulmonary sarcomatoid carcinoma (PSC), an uncommon and life-threatening family of non-small cell lung cancers. Methods: Ninety-eight PSCs were assessed for MNNG HOS Transforming gene (MET) and anaplastic lymphoma receptor tyrosine kinase gene (ALK) status by fluorescence in situ hybridization (FISH) and for relevant protein expression by immunohistochemical analysis, also taking advantage of phosphorylated (p-) antibodies. Moreover, levels of ALK and MET mRNA were also determined by real-time polymerase chain reaction and Western blot analysis for downstream activation pathways involving p-MET, p-protein kinase B, p-mitogen-activated protein kinase, p-SRC proto-oncogene tyrosine-protein kinase, and p-focal adhesion kinase (p-FAK). Results: MET amplification was detected by FISH in 25 of 98 PSCs (25.6%) and ALK amplification (but not the relevant rearrangement) was found in 16 of 98 (16.3%), with all ALKamplified tumors also showing MET amplification (p <0.0001). Nine PSCs, however, showed MET amplification without any ALK gene alteration. ALK protein expression was always lacking, whereas MET and p-MET were confined to the relevant amplified tumors only. Increased levels of ALK and MET mRNA were detectable in tumors with no direct relationship between mRNA content, protein expression, or alterations detected by FISH. Western blot assays showed complete activation of downstream signal pathways up to p-SRC proto-oncogene tyrosine-protein kinase, and p-focal adhesion kinase recruitment in MET and ALK-coamplified tumors only, whereas isolated MET amplification, MET and ALK borderline amplification (5%-10% of tumor cells with 15 copies of the relevant gene), or negative tumors showing eusomy or chromosome polysomy were confined to p-mitogen-activated protein kinase, p-protein kinase B, and/or p-MET activation. Multivariate survival analysis pushed a higher percentage of MET altered cells or a higher value of MET copy gain per cell to marginally emerge for overall survival (p = 0.140) and disease-free survival (p = 0.060), respectively. Conclusions: ALK and MET seemed to act as synergistic, nonrandom coactivators of downstream signal when coamplified in a subset of patients with PSC, thus likely suggesting a combined mechanism of oncogene addiction. These alterations could be a suitable target for therapy based on specific inhibitors.

Original languageEnglish
Pages (from-to)718-728
Number of pages11
JournalJournal of Thoracic Oncology
Volume11
Issue number5
DOIs
Publication statusPublished - 2016

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Lung Neoplasms
Carcinoma
Proto-Oncogene Proteins
Fluorescence In Situ Hybridization
Focal Adhesion Protein-Tyrosine Kinases
Neoplasms
Proto-Oncogene Proteins c-akt
Mitogen-Activated Protein Kinases
Oncogenes
Protein-Tyrosine Kinases
Messenger RNA
Therapeutics
Western Blotting
Genes
Activation Analysis
Methylnitronitrosoguanidine
Lung
Proteins
Survival Analysis
Non-Small Cell Lung Carcinoma

Keywords

  • ALK
  • FISH
  • Lung
  • MET
  • Sarcomatoid carcinoma
  • Survival

ASJC Scopus subject areas

  • Oncology
  • Pulmonary and Respiratory Medicine

Cite this

@article{5b1570564d9c4b24a60d0ae59382c6c1,
title = "Synergistic activation upon MET and ALK coamplification sustains targeted therapy in sarcomatoid Carcinoma, a deadly subtype of lung cancer",
abstract = "Introduction: Genetic alterations suitable for targeted therapy are poorly known issues in pulmonary sarcomatoid carcinoma (PSC), an uncommon and life-threatening family of non-small cell lung cancers. Methods: Ninety-eight PSCs were assessed for MNNG HOS Transforming gene (MET) and anaplastic lymphoma receptor tyrosine kinase gene (ALK) status by fluorescence in situ hybridization (FISH) and for relevant protein expression by immunohistochemical analysis, also taking advantage of phosphorylated (p-) antibodies. Moreover, levels of ALK and MET mRNA were also determined by real-time polymerase chain reaction and Western blot analysis for downstream activation pathways involving p-MET, p-protein kinase B, p-mitogen-activated protein kinase, p-SRC proto-oncogene tyrosine-protein kinase, and p-focal adhesion kinase (p-FAK). Results: MET amplification was detected by FISH in 25 of 98 PSCs (25.6{\%}) and ALK amplification (but not the relevant rearrangement) was found in 16 of 98 (16.3{\%}), with all ALKamplified tumors also showing MET amplification (p <0.0001). Nine PSCs, however, showed MET amplification without any ALK gene alteration. ALK protein expression was always lacking, whereas MET and p-MET were confined to the relevant amplified tumors only. Increased levels of ALK and MET mRNA were detectable in tumors with no direct relationship between mRNA content, protein expression, or alterations detected by FISH. Western blot assays showed complete activation of downstream signal pathways up to p-SRC proto-oncogene tyrosine-protein kinase, and p-focal adhesion kinase recruitment in MET and ALK-coamplified tumors only, whereas isolated MET amplification, MET and ALK borderline amplification (5{\%}-10{\%} of tumor cells with 15 copies of the relevant gene), or negative tumors showing eusomy or chromosome polysomy were confined to p-mitogen-activated protein kinase, p-protein kinase B, and/or p-MET activation. Multivariate survival analysis pushed a higher percentage of MET altered cells or a higher value of MET copy gain per cell to marginally emerge for overall survival (p = 0.140) and disease-free survival (p = 0.060), respectively. Conclusions: ALK and MET seemed to act as synergistic, nonrandom coactivators of downstream signal when coamplified in a subset of patients with PSC, thus likely suggesting a combined mechanism of oncogene addiction. These alterations could be a suitable target for therapy based on specific inhibitors.",
keywords = "ALK, FISH, Lung, MET, Sarcomatoid carcinoma, Survival",
author = "Giuseppe Pelosi and Patrizia Gasparini and Davide Conte and Alessandra Fabbri and Federica Perrone and Elena Tamborini and Serenella Pupa and Valentina Ciravolo and Roberto Caserini and Giulio Rossi and Alberto Cavazza and M. Papotti and Yukio Nakatani and Patrick Maisonneuve and Ugo Pastorino and Gabriella Sozzi",
year = "2016",
doi = "10.1016/j.jtho.2016.01.009",
language = "English",
volume = "11",
pages = "718--728",
journal = "Journal of Thoracic Oncology",
issn = "1556-0864",
publisher = "Elsevier Inc.",
number = "5",

}

TY - JOUR

T1 - Synergistic activation upon MET and ALK coamplification sustains targeted therapy in sarcomatoid Carcinoma, a deadly subtype of lung cancer

AU - Pelosi, Giuseppe

AU - Gasparini, Patrizia

AU - Conte, Davide

AU - Fabbri, Alessandra

AU - Perrone, Federica

AU - Tamborini, Elena

AU - Pupa, Serenella

AU - Ciravolo, Valentina

AU - Caserini, Roberto

AU - Rossi, Giulio

AU - Cavazza, Alberto

AU - Papotti, M.

AU - Nakatani, Yukio

AU - Maisonneuve, Patrick

AU - Pastorino, Ugo

AU - Sozzi, Gabriella

PY - 2016

Y1 - 2016

N2 - Introduction: Genetic alterations suitable for targeted therapy are poorly known issues in pulmonary sarcomatoid carcinoma (PSC), an uncommon and life-threatening family of non-small cell lung cancers. Methods: Ninety-eight PSCs were assessed for MNNG HOS Transforming gene (MET) and anaplastic lymphoma receptor tyrosine kinase gene (ALK) status by fluorescence in situ hybridization (FISH) and for relevant protein expression by immunohistochemical analysis, also taking advantage of phosphorylated (p-) antibodies. Moreover, levels of ALK and MET mRNA were also determined by real-time polymerase chain reaction and Western blot analysis for downstream activation pathways involving p-MET, p-protein kinase B, p-mitogen-activated protein kinase, p-SRC proto-oncogene tyrosine-protein kinase, and p-focal adhesion kinase (p-FAK). Results: MET amplification was detected by FISH in 25 of 98 PSCs (25.6%) and ALK amplification (but not the relevant rearrangement) was found in 16 of 98 (16.3%), with all ALKamplified tumors also showing MET amplification (p <0.0001). Nine PSCs, however, showed MET amplification without any ALK gene alteration. ALK protein expression was always lacking, whereas MET and p-MET were confined to the relevant amplified tumors only. Increased levels of ALK and MET mRNA were detectable in tumors with no direct relationship between mRNA content, protein expression, or alterations detected by FISH. Western blot assays showed complete activation of downstream signal pathways up to p-SRC proto-oncogene tyrosine-protein kinase, and p-focal adhesion kinase recruitment in MET and ALK-coamplified tumors only, whereas isolated MET amplification, MET and ALK borderline amplification (5%-10% of tumor cells with 15 copies of the relevant gene), or negative tumors showing eusomy or chromosome polysomy were confined to p-mitogen-activated protein kinase, p-protein kinase B, and/or p-MET activation. Multivariate survival analysis pushed a higher percentage of MET altered cells or a higher value of MET copy gain per cell to marginally emerge for overall survival (p = 0.140) and disease-free survival (p = 0.060), respectively. Conclusions: ALK and MET seemed to act as synergistic, nonrandom coactivators of downstream signal when coamplified in a subset of patients with PSC, thus likely suggesting a combined mechanism of oncogene addiction. These alterations could be a suitable target for therapy based on specific inhibitors.

AB - Introduction: Genetic alterations suitable for targeted therapy are poorly known issues in pulmonary sarcomatoid carcinoma (PSC), an uncommon and life-threatening family of non-small cell lung cancers. Methods: Ninety-eight PSCs were assessed for MNNG HOS Transforming gene (MET) and anaplastic lymphoma receptor tyrosine kinase gene (ALK) status by fluorescence in situ hybridization (FISH) and for relevant protein expression by immunohistochemical analysis, also taking advantage of phosphorylated (p-) antibodies. Moreover, levels of ALK and MET mRNA were also determined by real-time polymerase chain reaction and Western blot analysis for downstream activation pathways involving p-MET, p-protein kinase B, p-mitogen-activated protein kinase, p-SRC proto-oncogene tyrosine-protein kinase, and p-focal adhesion kinase (p-FAK). Results: MET amplification was detected by FISH in 25 of 98 PSCs (25.6%) and ALK amplification (but not the relevant rearrangement) was found in 16 of 98 (16.3%), with all ALKamplified tumors also showing MET amplification (p <0.0001). Nine PSCs, however, showed MET amplification without any ALK gene alteration. ALK protein expression was always lacking, whereas MET and p-MET were confined to the relevant amplified tumors only. Increased levels of ALK and MET mRNA were detectable in tumors with no direct relationship between mRNA content, protein expression, or alterations detected by FISH. Western blot assays showed complete activation of downstream signal pathways up to p-SRC proto-oncogene tyrosine-protein kinase, and p-focal adhesion kinase recruitment in MET and ALK-coamplified tumors only, whereas isolated MET amplification, MET and ALK borderline amplification (5%-10% of tumor cells with 15 copies of the relevant gene), or negative tumors showing eusomy or chromosome polysomy were confined to p-mitogen-activated protein kinase, p-protein kinase B, and/or p-MET activation. Multivariate survival analysis pushed a higher percentage of MET altered cells or a higher value of MET copy gain per cell to marginally emerge for overall survival (p = 0.140) and disease-free survival (p = 0.060), respectively. Conclusions: ALK and MET seemed to act as synergistic, nonrandom coactivators of downstream signal when coamplified in a subset of patients with PSC, thus likely suggesting a combined mechanism of oncogene addiction. These alterations could be a suitable target for therapy based on specific inhibitors.

KW - ALK

KW - FISH

KW - Lung

KW - MET

KW - Sarcomatoid carcinoma

KW - Survival

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UR - http://www.scopus.com/inward/citedby.url?scp=84969749877&partnerID=8YFLogxK

U2 - 10.1016/j.jtho.2016.01.009

DO - 10.1016/j.jtho.2016.01.009

M3 - Article

VL - 11

SP - 718

EP - 728

JO - Journal of Thoracic Oncology

JF - Journal of Thoracic Oncology

SN - 1556-0864

IS - 5

ER -