Identification and validation of DOCK4 as a potential biomarker for risk of bone metastasis development in patients with early breast cancer

Jules A Westbrook, Steven L Wood, David Cairns, Kathryn McMahon, Renu Gahlaut, Helene Thygesen, Mike Shires, Stephanie Roberts, Helen Marshall, Maria R Oliva, Mark J Dunning, Andrew M Hanby, Peter J Selby, Valerie Speirs, Georgia Mavria, Robert E. Coleman, Janet E Brown (Corresponding Author)

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Skeletal metastasis occurs in around 75% of advanced breast cancers, with the disease incurable once cancer cells disseminate to bone, but there remains an unmet need for biomarkers to identify patients at high risk of bone recurrence. This study aimed to identify such a biomarker and to assess its utility in predicting response to adjuvant zoledronic acid.

We used quantitative proteomics (SILAC‐MS), to compare protein expression in a bone‐homed variant (BM1) of the human breast cancer cell line MDA‐MB‐231 with parental non‐bone‐homing cells to identify novel biomarkers for risk of subsequent bone metastasis in early breast cancer. SILAC‐MS showed that Dedicator of cytokinesis protein 4 (DOCK4) was upregulated in bone‐homing BM1 cells, confirmed by Western blotting. BM1 cells also had enhanced invasive ability compared with parental cells which could be reduced by DOCK4‐shRNA.

In a training Tissue Microarray (TMA) comprising 345 patients with early breast cancer, immunohistochemistry followed by Cox regression revealed that high DOCK4 expression correlated with histological grade (p=0.004) but not oestrogen receptor status (p=0.19) or lymph node involvement (p=0.15). A clinical validation TMA used tissue samples and the clinical database from the large AZURE adjuvant study (n=689). Adjusted Cox regression analyses showed that high DOCK4 expression in the control arm (no zoledronic acid) was significantly prognostic for first recurrence in bone (HR 2.13, 95%CI 1.06‐4.30, p=0.034). No corresponding association was found in patients who received zoledronic acid (HR 0.812, 95%CI 0.176‐3.76, p=0.790), suggesting that treatment with zoledronic acid may counteract the higher risk for bone relapse from high DOCK4‐expressing tumours.

High DOCK4 expression was not associated with metastasis to non‐skeletal sites when these were assessed collectively. In conclusion, high DOCK4 in early breast cancer is significantly associated with aggressive disease and with future bone metastasis and is a potentially useful biomarker for subsequent bone metastasis risk.

This article is protected by copyright. All rights reserved.
Original languageEnglish
Pages (from-to)381-391
Number of pages11
JournalThe Journal of pathology
Volume247
Issue number3
Early online date14 Nov 2018
DOIs
Publication statusPublished - Mar 2019

Fingerprint

zoledronic acid
Cytokinesis
Bone Development
Biomarkers
Breast Neoplasms
Neoplasm Metastasis
Bone and Bones
Proteins
Recurrence
Estrogen Receptors
Proteomics
Neoplasms
Lymph Nodes
Western Blotting
Immunohistochemistry
Regression Analysis
Databases
Cell Line

Keywords

  • DOCK4
  • bone metastasis
  • breast cancer
  • biomarker
  • proteomics
  • AMINO-ACIDS
  • CELL-MIGRATION
  • TUMOR
  • PROTEINS
  • CULTURE
  • EXPRESSION
  • ADJUVANT ZOLEDRONIC ACID

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Identification and validation of DOCK4 as a potential biomarker for risk of bone metastasis development in patients with early breast cancer. / Westbrook, Jules A; Wood, Steven L; Cairns, David; McMahon, Kathryn; Gahlaut, Renu ; Thygesen, Helene; Shires, Mike; Roberts, Stephanie ; Marshall, Helen ; Oliva, Maria R; Dunning, Mark J; Hanby, Andrew M; Selby, Peter J; Speirs, Valerie; Mavria, Georgia; Coleman, Robert E.; Brown, Janet E (Corresponding Author).

In: The Journal of pathology, Vol. 247, No. 3, 03.2019, p. 381-391.

Research output: Contribution to journalArticle

Westbrook, JA, Wood, SL, Cairns, D, McMahon, K, Gahlaut, R, Thygesen, H, Shires, M, Roberts, S, Marshall, H, Oliva, MR, Dunning, MJ, Hanby, AM, Selby, PJ, Speirs, V, Mavria, G, Coleman, RE & Brown, JE 2019, 'Identification and validation of DOCK4 as a potential biomarker for risk of bone metastasis development in patients with early breast cancer', The Journal of pathology, vol. 247, no. 3, pp. 381-391. https://doi.org/10.1002/path.5197
Westbrook, Jules A ; Wood, Steven L ; Cairns, David ; McMahon, Kathryn ; Gahlaut, Renu ; Thygesen, Helene ; Shires, Mike ; Roberts, Stephanie ; Marshall, Helen ; Oliva, Maria R ; Dunning, Mark J ; Hanby, Andrew M ; Selby, Peter J ; Speirs, Valerie ; Mavria, Georgia ; Coleman, Robert E. ; Brown, Janet E. / Identification and validation of DOCK4 as a potential biomarker for risk of bone metastasis development in patients with early breast cancer. In: The Journal of pathology. 2019 ; Vol. 247, No. 3. pp. 381-391.
@article{95c031f5beef435f95df7091e05885fa,
title = "Identification and validation of DOCK4 as a potential biomarker for risk of bone metastasis development in patients with early breast cancer",
abstract = "Skeletal metastasis occurs in around 75{\%} of advanced breast cancers, with the disease incurable once cancer cells disseminate to bone, but there remains an unmet need for biomarkers to identify patients at high risk of bone recurrence. This study aimed to identify such a biomarker and to assess its utility in predicting response to adjuvant zoledronic acid.We used quantitative proteomics (SILAC‐MS), to compare protein expression in a bone‐homed variant (BM1) of the human breast cancer cell line MDA‐MB‐231 with parental non‐bone‐homing cells to identify novel biomarkers for risk of subsequent bone metastasis in early breast cancer. SILAC‐MS showed that Dedicator of cytokinesis protein 4 (DOCK4) was upregulated in bone‐homing BM1 cells, confirmed by Western blotting. BM1 cells also had enhanced invasive ability compared with parental cells which could be reduced by DOCK4‐shRNA.In a training Tissue Microarray (TMA) comprising 345 patients with early breast cancer, immunohistochemistry followed by Cox regression revealed that high DOCK4 expression correlated with histological grade (p=0.004) but not oestrogen receptor status (p=0.19) or lymph node involvement (p=0.15). A clinical validation TMA used tissue samples and the clinical database from the large AZURE adjuvant study (n=689). Adjusted Cox regression analyses showed that high DOCK4 expression in the control arm (no zoledronic acid) was significantly prognostic for first recurrence in bone (HR 2.13, 95{\%}CI 1.06‐4.30, p=0.034). No corresponding association was found in patients who received zoledronic acid (HR 0.812, 95{\%}CI 0.176‐3.76, p=0.790), suggesting that treatment with zoledronic acid may counteract the higher risk for bone relapse from high DOCK4‐expressing tumours.High DOCK4 expression was not associated with metastasis to non‐skeletal sites when these were assessed collectively. In conclusion, high DOCK4 in early breast cancer is significantly associated with aggressive disease and with future bone metastasis and is a potentially useful biomarker for subsequent bone metastasis risk.This article is protected by copyright. All rights reserved.",
keywords = "DOCK4, bone metastasis, breast cancer, biomarker, proteomics, AMINO-ACIDS, CELL-MIGRATION, TUMOR, PROTEINS, CULTURE, EXPRESSION, ADJUVANT ZOLEDRONIC ACID",
author = "Westbrook, {Jules A} and Wood, {Steven L} and David Cairns and Kathryn McMahon and Renu Gahlaut and Helene Thygesen and Mike Shires and Stephanie Roberts and Helen Marshall and Oliva, {Maria R} and Dunning, {Mark J} and Hanby, {Andrew M} and Selby, {Peter J} and Valerie Speirs and Georgia Mavria and Coleman, {Robert E.} and Brown, {Janet E}",
note = "Funding Information Cancer Research UK. Grant Number: C18605/A 10048 Yorkshire Cancer Research. Grant Number: S315 Breast Cancer Now. Grant Numbers: 2012MaySP047, 2016MayPR74 UK Medical Research Council. Grant Number: G0802416",
year = "2019",
month = "3",
doi = "10.1002/path.5197",
language = "English",
volume = "247",
pages = "381--391",
journal = "The Journal of pathology",
issn = "0022-3417",
publisher = "Wiley",
number = "3",

}

TY - JOUR

T1 - Identification and validation of DOCK4 as a potential biomarker for risk of bone metastasis development in patients with early breast cancer

AU - Westbrook, Jules A

AU - Wood, Steven L

AU - Cairns, David

AU - McMahon, Kathryn

AU - Gahlaut, Renu

AU - Thygesen, Helene

AU - Shires, Mike

AU - Roberts, Stephanie

AU - Marshall, Helen

AU - Oliva, Maria R

AU - Dunning, Mark J

AU - Hanby, Andrew M

AU - Selby, Peter J

AU - Speirs, Valerie

AU - Mavria, Georgia

AU - Coleman, Robert E.

AU - Brown, Janet E

N1 - Funding Information Cancer Research UK. Grant Number: C18605/A 10048 Yorkshire Cancer Research. Grant Number: S315 Breast Cancer Now. Grant Numbers: 2012MaySP047, 2016MayPR74 UK Medical Research Council. Grant Number: G0802416

PY - 2019/3

Y1 - 2019/3

N2 - Skeletal metastasis occurs in around 75% of advanced breast cancers, with the disease incurable once cancer cells disseminate to bone, but there remains an unmet need for biomarkers to identify patients at high risk of bone recurrence. This study aimed to identify such a biomarker and to assess its utility in predicting response to adjuvant zoledronic acid.We used quantitative proteomics (SILAC‐MS), to compare protein expression in a bone‐homed variant (BM1) of the human breast cancer cell line MDA‐MB‐231 with parental non‐bone‐homing cells to identify novel biomarkers for risk of subsequent bone metastasis in early breast cancer. SILAC‐MS showed that Dedicator of cytokinesis protein 4 (DOCK4) was upregulated in bone‐homing BM1 cells, confirmed by Western blotting. BM1 cells also had enhanced invasive ability compared with parental cells which could be reduced by DOCK4‐shRNA.In a training Tissue Microarray (TMA) comprising 345 patients with early breast cancer, immunohistochemistry followed by Cox regression revealed that high DOCK4 expression correlated with histological grade (p=0.004) but not oestrogen receptor status (p=0.19) or lymph node involvement (p=0.15). A clinical validation TMA used tissue samples and the clinical database from the large AZURE adjuvant study (n=689). Adjusted Cox regression analyses showed that high DOCK4 expression in the control arm (no zoledronic acid) was significantly prognostic for first recurrence in bone (HR 2.13, 95%CI 1.06‐4.30, p=0.034). No corresponding association was found in patients who received zoledronic acid (HR 0.812, 95%CI 0.176‐3.76, p=0.790), suggesting that treatment with zoledronic acid may counteract the higher risk for bone relapse from high DOCK4‐expressing tumours.High DOCK4 expression was not associated with metastasis to non‐skeletal sites when these were assessed collectively. In conclusion, high DOCK4 in early breast cancer is significantly associated with aggressive disease and with future bone metastasis and is a potentially useful biomarker for subsequent bone metastasis risk.This article is protected by copyright. All rights reserved.

AB - Skeletal metastasis occurs in around 75% of advanced breast cancers, with the disease incurable once cancer cells disseminate to bone, but there remains an unmet need for biomarkers to identify patients at high risk of bone recurrence. This study aimed to identify such a biomarker and to assess its utility in predicting response to adjuvant zoledronic acid.We used quantitative proteomics (SILAC‐MS), to compare protein expression in a bone‐homed variant (BM1) of the human breast cancer cell line MDA‐MB‐231 with parental non‐bone‐homing cells to identify novel biomarkers for risk of subsequent bone metastasis in early breast cancer. SILAC‐MS showed that Dedicator of cytokinesis protein 4 (DOCK4) was upregulated in bone‐homing BM1 cells, confirmed by Western blotting. BM1 cells also had enhanced invasive ability compared with parental cells which could be reduced by DOCK4‐shRNA.In a training Tissue Microarray (TMA) comprising 345 patients with early breast cancer, immunohistochemistry followed by Cox regression revealed that high DOCK4 expression correlated with histological grade (p=0.004) but not oestrogen receptor status (p=0.19) or lymph node involvement (p=0.15). A clinical validation TMA used tissue samples and the clinical database from the large AZURE adjuvant study (n=689). Adjusted Cox regression analyses showed that high DOCK4 expression in the control arm (no zoledronic acid) was significantly prognostic for first recurrence in bone (HR 2.13, 95%CI 1.06‐4.30, p=0.034). No corresponding association was found in patients who received zoledronic acid (HR 0.812, 95%CI 0.176‐3.76, p=0.790), suggesting that treatment with zoledronic acid may counteract the higher risk for bone relapse from high DOCK4‐expressing tumours.High DOCK4 expression was not associated with metastasis to non‐skeletal sites when these were assessed collectively. In conclusion, high DOCK4 in early breast cancer is significantly associated with aggressive disease and with future bone metastasis and is a potentially useful biomarker for subsequent bone metastasis risk.This article is protected by copyright. All rights reserved.

KW - DOCK4

KW - bone metastasis

KW - breast cancer

KW - biomarker

KW - proteomics

KW - AMINO-ACIDS

KW - CELL-MIGRATION

KW - TUMOR

KW - PROTEINS

KW - CULTURE

KW - EXPRESSION

KW - ADJUVANT ZOLEDRONIC ACID

UR - http://www.scopus.com/inward/record.url?scp=85060606747&partnerID=8YFLogxK

UR - http://www.mendeley.com/research/identification-validation-dock4-potential-biomarker-risk-bone-metastasis-development-patients-early

U2 - 10.1002/path.5197

DO - 10.1002/path.5197

M3 - Article

VL - 247

SP - 381

EP - 391

JO - The Journal of pathology

JF - The Journal of pathology

SN - 0022-3417

IS - 3

ER -