The role of c-MET/HGF signaling as a critical mediator of an invasive and resistant phenotype in colorectal cancer

Conor Bradley, Philip Dunne, Darragh McArt, Ken Arthur, Stephen McQuaid, Manuel Salto-Tellez, Patrick Johnston, Sandra Van Schaeybroeck

Research output: Chapter in Book/Report/Conference proceedingConference contribution

Abstract

The c-MET proto-oncogene is frequently overexpressed (50-60%), amplified (5-10%), and mutated (5%) in colorectal cancer (CRC). Hepatocyte growth factor (HGF)-induced c-MET activation has been linked with migration, survival, invasion, and resistance to targeted therapy, and has been suggested as a possible prognostic biomarker for CRC. We are currently investigating the role of c-MET and microenvironment derived HGF in the acquisition of an invasive and migratory phenotype, and resistance to targeted therapy in CRC.
In order to model CRC tumour cell invasion and metastasis, we have generated invasive subpopulations from CRC cells using Boyden Invasion chambers. Invasive cell lines were characterised for protein expression/activity by Western blotting, and analysed for migratory and invasive potential using the xCELLigence System (Roche). To model the CRC microenvironment, we have utilised a range of co-culture techniques with CRC cell lines and colon fibroblasts. c-MET expression in FFPE tissues was measured using IHC in a tissue microarray (TMA) derived from early stage CRC patients.
HCT116 and LoVo invasive subpopulations showed an EMT-like, mesenchymal, migratory/invasive phenotype. In addition, increased expression and activation levels of c-MET were found in these sublines, which was determined to be ligand independent. Inhibition of c-MET using RNAi abrogated both basal and HGF-induced migration and invasion in CRC cell lines. Co-culture of CRC cells with HGF-expressing colon-derived fibroblasts leads to activation of c-MET, inducing CRC migration, invasion, and resistance to MEK inhibition, and this phenotype could be diminished using a HGF neutralising antibody. Significant increased expression levels of c-MET were also found in a CRC TMA compared to matched normal tissues, and also at the invasive edge of some CRC tumours, where it may be driving invasive biology.
The identification of key pathways driving metastasis has a huge potential to change treatment strategies in CRC. We intend to further investigate the role of c-MET in regulation of migration, invasion, and resistance to therapy in CRC. We hypothesise that patients with high tumoural c-MET at the invasive front may benefit in particular from anti- c-MET therapies.
Original languageEnglish
Title of host publicationThe role of c-MET/HGF signaling as a critical mediator of an invasive and resistant phenotype in colorectal cancer
PublisherProceedings: AACR Annual Meeting 2015; April 18-22, Philadelphia
VolumeAbstract 4018
Publication statusPublished - 2015

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Hepatocyte Growth Factor
Colorectal Neoplasms
Phenotype
Coculture Techniques
Cell Line
Colon
Fibroblasts
Neoplasm Metastasis
Therapeutics
Culture Techniques
Tumor Microenvironment
Proto-Oncogenes
Mitogen-Activated Protein Kinase Kinases
RNA Interference
Neutralizing Antibodies

Cite this

Bradley, C., Dunne, P., McArt, D., Arthur, K., McQuaid, S., Salto-Tellez, M., ... Van Schaeybroeck, S. (2015). The role of c-MET/HGF signaling as a critical mediator of an invasive and resistant phenotype in colorectal cancer. In The role of c-MET/HGF signaling as a critical mediator of an invasive and resistant phenotype in colorectal cancer (Vol. Abstract 4018). Proceedings: AACR Annual Meeting 2015; April 18-22, Philadelphia.
Bradley, Conor ; Dunne, Philip ; McArt, Darragh ; Arthur, Ken ; McQuaid, Stephen ; Salto-Tellez, Manuel ; Johnston, Patrick ; Van Schaeybroeck, Sandra. / The role of c-MET/HGF signaling as a critical mediator of an invasive and resistant phenotype in colorectal cancer. The role of c-MET/HGF signaling as a critical mediator of an invasive and resistant phenotype in colorectal cancer. Vol. Abstract 4018 Proceedings: AACR Annual Meeting 2015; April 18-22, Philadelphia, 2015.
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title = "The role of c-MET/HGF signaling as a critical mediator of an invasive and resistant phenotype in colorectal cancer",
abstract = "The c-MET proto-oncogene is frequently overexpressed (50-60{\%}), amplified (5-10{\%}), and mutated (5{\%}) in colorectal cancer (CRC). Hepatocyte growth factor (HGF)-induced c-MET activation has been linked with migration, survival, invasion, and resistance to targeted therapy, and has been suggested as a possible prognostic biomarker for CRC. We are currently investigating the role of c-MET and microenvironment derived HGF in the acquisition of an invasive and migratory phenotype, and resistance to targeted therapy in CRC.In order to model CRC tumour cell invasion and metastasis, we have generated invasive subpopulations from CRC cells using Boyden Invasion chambers. Invasive cell lines were characterised for protein expression/activity by Western blotting, and analysed for migratory and invasive potential using the xCELLigence System (Roche). To model the CRC microenvironment, we have utilised a range of co-culture techniques with CRC cell lines and colon fibroblasts. c-MET expression in FFPE tissues was measured using IHC in a tissue microarray (TMA) derived from early stage CRC patients.HCT116 and LoVo invasive subpopulations showed an EMT-like, mesenchymal, migratory/invasive phenotype. In addition, increased expression and activation levels of c-MET were found in these sublines, which was determined to be ligand independent. Inhibition of c-MET using RNAi abrogated both basal and HGF-induced migration and invasion in CRC cell lines. Co-culture of CRC cells with HGF-expressing colon-derived fibroblasts leads to activation of c-MET, inducing CRC migration, invasion, and resistance to MEK inhibition, and this phenotype could be diminished using a HGF neutralising antibody. Significant increased expression levels of c-MET were also found in a CRC TMA compared to matched normal tissues, and also at the invasive edge of some CRC tumours, where it may be driving invasive biology.The identification of key pathways driving metastasis has a huge potential to change treatment strategies in CRC. We intend to further investigate the role of c-MET in regulation of migration, invasion, and resistance to therapy in CRC. We hypothesise that patients with high tumoural c-MET at the invasive front may benefit in particular from anti- c-MET therapies.",
author = "Conor Bradley and Philip Dunne and Darragh McArt and Ken Arthur and Stephen McQuaid and Manuel Salto-Tellez and Patrick Johnston and {Van Schaeybroeck}, Sandra",
year = "2015",
language = "English",
volume = "Abstract 4018",
booktitle = "The role of c-MET/HGF signaling as a critical mediator of an invasive and resistant phenotype in colorectal cancer",
publisher = "Proceedings: AACR Annual Meeting 2015; April 18-22, Philadelphia",

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Bradley, C, Dunne, P, McArt, D, Arthur, K, McQuaid, S, Salto-Tellez, M, Johnston, P & Van Schaeybroeck, S 2015, The role of c-MET/HGF signaling as a critical mediator of an invasive and resistant phenotype in colorectal cancer. in The role of c-MET/HGF signaling as a critical mediator of an invasive and resistant phenotype in colorectal cancer. vol. Abstract 4018, Proceedings: AACR Annual Meeting 2015; April 18-22, Philadelphia.

The role of c-MET/HGF signaling as a critical mediator of an invasive and resistant phenotype in colorectal cancer. / Bradley, Conor; Dunne, Philip; McArt, Darragh; Arthur, Ken; McQuaid, Stephen; Salto-Tellez, Manuel; Johnston, Patrick; Van Schaeybroeck, Sandra.

The role of c-MET/HGF signaling as a critical mediator of an invasive and resistant phenotype in colorectal cancer. Vol. Abstract 4018 Proceedings: AACR Annual Meeting 2015; April 18-22, Philadelphia, 2015.

Research output: Chapter in Book/Report/Conference proceedingConference contribution

TY - GEN

T1 - The role of c-MET/HGF signaling as a critical mediator of an invasive and resistant phenotype in colorectal cancer

AU - Bradley, Conor

AU - Dunne, Philip

AU - McArt, Darragh

AU - Arthur, Ken

AU - McQuaid, Stephen

AU - Salto-Tellez, Manuel

AU - Johnston, Patrick

AU - Van Schaeybroeck, Sandra

PY - 2015

Y1 - 2015

N2 - The c-MET proto-oncogene is frequently overexpressed (50-60%), amplified (5-10%), and mutated (5%) in colorectal cancer (CRC). Hepatocyte growth factor (HGF)-induced c-MET activation has been linked with migration, survival, invasion, and resistance to targeted therapy, and has been suggested as a possible prognostic biomarker for CRC. We are currently investigating the role of c-MET and microenvironment derived HGF in the acquisition of an invasive and migratory phenotype, and resistance to targeted therapy in CRC.In order to model CRC tumour cell invasion and metastasis, we have generated invasive subpopulations from CRC cells using Boyden Invasion chambers. Invasive cell lines were characterised for protein expression/activity by Western blotting, and analysed for migratory and invasive potential using the xCELLigence System (Roche). To model the CRC microenvironment, we have utilised a range of co-culture techniques with CRC cell lines and colon fibroblasts. c-MET expression in FFPE tissues was measured using IHC in a tissue microarray (TMA) derived from early stage CRC patients.HCT116 and LoVo invasive subpopulations showed an EMT-like, mesenchymal, migratory/invasive phenotype. In addition, increased expression and activation levels of c-MET were found in these sublines, which was determined to be ligand independent. Inhibition of c-MET using RNAi abrogated both basal and HGF-induced migration and invasion in CRC cell lines. Co-culture of CRC cells with HGF-expressing colon-derived fibroblasts leads to activation of c-MET, inducing CRC migration, invasion, and resistance to MEK inhibition, and this phenotype could be diminished using a HGF neutralising antibody. Significant increased expression levels of c-MET were also found in a CRC TMA compared to matched normal tissues, and also at the invasive edge of some CRC tumours, where it may be driving invasive biology.The identification of key pathways driving metastasis has a huge potential to change treatment strategies in CRC. We intend to further investigate the role of c-MET in regulation of migration, invasion, and resistance to therapy in CRC. We hypothesise that patients with high tumoural c-MET at the invasive front may benefit in particular from anti- c-MET therapies.

AB - The c-MET proto-oncogene is frequently overexpressed (50-60%), amplified (5-10%), and mutated (5%) in colorectal cancer (CRC). Hepatocyte growth factor (HGF)-induced c-MET activation has been linked with migration, survival, invasion, and resistance to targeted therapy, and has been suggested as a possible prognostic biomarker for CRC. We are currently investigating the role of c-MET and microenvironment derived HGF in the acquisition of an invasive and migratory phenotype, and resistance to targeted therapy in CRC.In order to model CRC tumour cell invasion and metastasis, we have generated invasive subpopulations from CRC cells using Boyden Invasion chambers. Invasive cell lines were characterised for protein expression/activity by Western blotting, and analysed for migratory and invasive potential using the xCELLigence System (Roche). To model the CRC microenvironment, we have utilised a range of co-culture techniques with CRC cell lines and colon fibroblasts. c-MET expression in FFPE tissues was measured using IHC in a tissue microarray (TMA) derived from early stage CRC patients.HCT116 and LoVo invasive subpopulations showed an EMT-like, mesenchymal, migratory/invasive phenotype. In addition, increased expression and activation levels of c-MET were found in these sublines, which was determined to be ligand independent. Inhibition of c-MET using RNAi abrogated both basal and HGF-induced migration and invasion in CRC cell lines. Co-culture of CRC cells with HGF-expressing colon-derived fibroblasts leads to activation of c-MET, inducing CRC migration, invasion, and resistance to MEK inhibition, and this phenotype could be diminished using a HGF neutralising antibody. Significant increased expression levels of c-MET were also found in a CRC TMA compared to matched normal tissues, and also at the invasive edge of some CRC tumours, where it may be driving invasive biology.The identification of key pathways driving metastasis has a huge potential to change treatment strategies in CRC. We intend to further investigate the role of c-MET in regulation of migration, invasion, and resistance to therapy in CRC. We hypothesise that patients with high tumoural c-MET at the invasive front may benefit in particular from anti- c-MET therapies.

M3 - Conference contribution

VL - Abstract 4018

BT - The role of c-MET/HGF signaling as a critical mediator of an invasive and resistant phenotype in colorectal cancer

PB - Proceedings: AACR Annual Meeting 2015; April 18-22, Philadelphia

ER -

Bradley C, Dunne P, McArt D, Arthur K, McQuaid S, Salto-Tellez M et al. The role of c-MET/HGF signaling as a critical mediator of an invasive and resistant phenotype in colorectal cancer. In The role of c-MET/HGF signaling as a critical mediator of an invasive and resistant phenotype in colorectal cancer. Vol. Abstract 4018. Proceedings: AACR Annual Meeting 2015; April 18-22, Philadelphia. 2015