Quantitative assessment of the cell surface proteome to identify novel therapeutic targets in cholangiocarcinoma

Barney Stephenson; Neil Shimwell; Elizabeth Humphreys; Douglas Ward; David Adams; Ashley Martin; Simon Afford

doi:10.1016/S0140-6736(15)60409-3

Quantitative assessment of the cell surface proteome to identify novel therapeutic targets in cholangiocarcinoma

Barney Stephenson, Neil Shimwell, Elizabeth Humphreys, Douglas Ward, David Adams, Ashley Martin, Simon Afford

Research output: Chapter in Book/Report/Conference proceeding › Conference contribution

Abstract

BACKGROUND: Cholangiocarcinoma has a high mortality and morbidity. Median survival is less than 6 months. Surgical resection is appropriate in certain circumstances. Because distal cholangiocarcinoma is difficult to distinguish from pancreatic cancers, patients might not receive optimum therapy. Proteomics is the study of complex cellular proteins using mass spectrometry. The aim of this study was to determine the constituent proteins on the cell surface of a model of cholangiocarcinoma.

METHODS: A sample preparation technique to enrich for cell surface proteins of the intrahepatic cholangiocarcinoma cell line CC-SW-1 was developed by modifying a NeutrAvidin-biotin system. After isolation, trypsin digestion, and purification, peptides were fractionated for tandem mass spectrometry before being analysed with the NCBInr database and the Mascot search algorithm. Results were confirmed by immunohistochemistry using a peroxidase detection technique on paraffin-embedded sections from resected specimens.

FINDINGS: Peptide enrichment was confirmed by electrophoresis. 862 proteins were consistently expressed between samples (n=3). 271 of these proteins were attributed only to the cell surface. They included proteins used clinically for staging disease (cytokeratin 19 [CK19]), identifying cancer stem cells (epithelial cell adhesion molecule [EpCAM], neural cell adhesion molecule [NCAM], epithelial growth factor receptor [EGFR]), and indicating potential for differentiation (Frozzled receptor, Notch pathway). Novel markers from the tumour necrosis factor (TNF) receptor superfamily were also identified. Immunohistochemistry confirmed these findings.

INTERPRETATION: The results from this surface proteomic profiling could help to identify novel therapeutic targets in cholangiocarcinoma. Further development of this technique could be translated to distinguish between distal cholangiocarcinoma and pancreatic cancers.

FUNDING: UK Medical Research Council.

Original language	English
Title of host publication	The Lancet
Subtitle of host publication	Spring Meeting for Clinician Scientists in Training 2015
Pages	S94
Volume	385 Suppl 1
DOIs	https://doi.org/10.1016/S0140-6736(15)60409-3
Publication status	Published - 26 Feb 2015
Event	Spring Meeting for Clinician Scientists in Training 2015 - Royal College of Physicians, London, United Kingdom Duration: 26 Feb 2015 → 26 Feb 2015

Conference

Conference	Spring Meeting for Clinician Scientists in Training 2015
Country/Territory	United Kingdom
City	London
Period	26/02/15 → 26/02/15

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/S0140-6736(15)60409-3

The Functional Significance of the Fn14/TWEAK Receptor-Ligand System in Ductular Reactive Cell Differentiation and Bile Ductule Neogenesis
Afford, S. & Adams, D.
Medical Research Council
1/05/12 → 30/04/15
Project: Research Councils

Cite this

@inproceedings{5e34de2ec3f14549ba5fc3160f56ffc6,

title = "Quantitative assessment of the cell surface proteome to identify novel therapeutic targets in cholangiocarcinoma",

abstract = "BACKGROUND: Cholangiocarcinoma has a high mortality and morbidity. Median survival is less than 6 months. Surgical resection is appropriate in certain circumstances. Because distal cholangiocarcinoma is difficult to distinguish from pancreatic cancers, patients might not receive optimum therapy. Proteomics is the study of complex cellular proteins using mass spectrometry. The aim of this study was to determine the constituent proteins on the cell surface of a model of cholangiocarcinoma.METHODS: A sample preparation technique to enrich for cell surface proteins of the intrahepatic cholangiocarcinoma cell line CC-SW-1 was developed by modifying a NeutrAvidin-biotin system. After isolation, trypsin digestion, and purification, peptides were fractionated for tandem mass spectrometry before being analysed with the NCBInr database and the Mascot search algorithm. Results were confirmed by immunohistochemistry using a peroxidase detection technique on paraffin-embedded sections from resected specimens.FINDINGS: Peptide enrichment was confirmed by electrophoresis. 862 proteins were consistently expressed between samples (n=3). 271 of these proteins were attributed only to the cell surface. They included proteins used clinically for staging disease (cytokeratin 19 [CK19]), identifying cancer stem cells (epithelial cell adhesion molecule [EpCAM], neural cell adhesion molecule [NCAM], epithelial growth factor receptor [EGFR]), and indicating potential for differentiation (Frozzled receptor, Notch pathway). Novel markers from the tumour necrosis factor (TNF) receptor superfamily were also identified. Immunohistochemistry confirmed these findings.INTERPRETATION: The results from this surface proteomic profiling could help to identify novel therapeutic targets in cholangiocarcinoma. Further development of this technique could be translated to distinguish between distal cholangiocarcinoma and pancreatic cancers.FUNDING: UK Medical Research Council.",

author = "Barney Stephenson and Neil Shimwell and Elizabeth Humphreys and Douglas Ward and David Adams and Ashley Martin and Simon Afford",

year = "2015",

month = feb,

day = "26",

doi = "10.1016/S0140-6736(15)60409-3",

language = "English",

volume = "385 Suppl 1",

pages = "S94",

booktitle = "The Lancet",

note = "Spring Meeting for Clinician Scientists in Training 2015 ; Conference date: 26-02-2015 Through 26-02-2015",

}

Stephenson, B, Shimwell, N, Humphreys, E, Ward, D , Adams, D, Martin, A & Afford, S 2015, Quantitative assessment of the cell surface proteome to identify novel therapeutic targets in cholangiocarcinoma. in The Lancet: Spring Meeting for Clinician Scientists in Training 2015. vol. 385 Suppl 1, pp. S94, Spring Meeting for Clinician Scientists in Training 2015, London, United Kingdom, 26/02/15. https://doi.org/10.1016/S0140-6736(15)60409-3

TY - GEN

T1 - Quantitative assessment of the cell surface proteome to identify novel therapeutic targets in cholangiocarcinoma

AU - Stephenson, Barney

AU - Shimwell, Neil

AU - Humphreys, Elizabeth

AU - Ward, Douglas

AU - Adams, David

AU - Martin, Ashley

AU - Afford, Simon

PY - 2015/2/26

Y1 - 2015/2/26

N2 - BACKGROUND: Cholangiocarcinoma has a high mortality and morbidity. Median survival is less than 6 months. Surgical resection is appropriate in certain circumstances. Because distal cholangiocarcinoma is difficult to distinguish from pancreatic cancers, patients might not receive optimum therapy. Proteomics is the study of complex cellular proteins using mass spectrometry. The aim of this study was to determine the constituent proteins on the cell surface of a model of cholangiocarcinoma.METHODS: A sample preparation technique to enrich for cell surface proteins of the intrahepatic cholangiocarcinoma cell line CC-SW-1 was developed by modifying a NeutrAvidin-biotin system. After isolation, trypsin digestion, and purification, peptides were fractionated for tandem mass spectrometry before being analysed with the NCBInr database and the Mascot search algorithm. Results were confirmed by immunohistochemistry using a peroxidase detection technique on paraffin-embedded sections from resected specimens.FINDINGS: Peptide enrichment was confirmed by electrophoresis. 862 proteins were consistently expressed between samples (n=3). 271 of these proteins were attributed only to the cell surface. They included proteins used clinically for staging disease (cytokeratin 19 [CK19]), identifying cancer stem cells (epithelial cell adhesion molecule [EpCAM], neural cell adhesion molecule [NCAM], epithelial growth factor receptor [EGFR]), and indicating potential for differentiation (Frozzled receptor, Notch pathway). Novel markers from the tumour necrosis factor (TNF) receptor superfamily were also identified. Immunohistochemistry confirmed these findings.INTERPRETATION: The results from this surface proteomic profiling could help to identify novel therapeutic targets in cholangiocarcinoma. Further development of this technique could be translated to distinguish between distal cholangiocarcinoma and pancreatic cancers.FUNDING: UK Medical Research Council.

AB - BACKGROUND: Cholangiocarcinoma has a high mortality and morbidity. Median survival is less than 6 months. Surgical resection is appropriate in certain circumstances. Because distal cholangiocarcinoma is difficult to distinguish from pancreatic cancers, patients might not receive optimum therapy. Proteomics is the study of complex cellular proteins using mass spectrometry. The aim of this study was to determine the constituent proteins on the cell surface of a model of cholangiocarcinoma.METHODS: A sample preparation technique to enrich for cell surface proteins of the intrahepatic cholangiocarcinoma cell line CC-SW-1 was developed by modifying a NeutrAvidin-biotin system. After isolation, trypsin digestion, and purification, peptides were fractionated for tandem mass spectrometry before being analysed with the NCBInr database and the Mascot search algorithm. Results were confirmed by immunohistochemistry using a peroxidase detection technique on paraffin-embedded sections from resected specimens.FINDINGS: Peptide enrichment was confirmed by electrophoresis. 862 proteins were consistently expressed between samples (n=3). 271 of these proteins were attributed only to the cell surface. They included proteins used clinically for staging disease (cytokeratin 19 [CK19]), identifying cancer stem cells (epithelial cell adhesion molecule [EpCAM], neural cell adhesion molecule [NCAM], epithelial growth factor receptor [EGFR]), and indicating potential for differentiation (Frozzled receptor, Notch pathway). Novel markers from the tumour necrosis factor (TNF) receptor superfamily were also identified. Immunohistochemistry confirmed these findings.INTERPRETATION: The results from this surface proteomic profiling could help to identify novel therapeutic targets in cholangiocarcinoma. Further development of this technique could be translated to distinguish between distal cholangiocarcinoma and pancreatic cancers.FUNDING: UK Medical Research Council.

U2 - 10.1016/S0140-6736(15)60409-3

DO - 10.1016/S0140-6736(15)60409-3

M3 - Conference contribution

C2 - 26312917

VL - 385 Suppl 1

SP - S94

BT - The Lancet

T2 - Spring Meeting for Clinician Scientists in Training 2015

Y2 - 26 February 2015 through 26 February 2015

ER -

Quantitative assessment of the cell surface proteome to identify novel therapeutic targets in cholangiocarcinoma

Abstract

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UN SDGs

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The Functional Significance of the Fn14/TWEAK Receptor-Ligand System in Ductular Reactive Cell Differentiation and Bile Ductule Neogenesis

Cite this