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 A 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

doi:10.1002/path.5197

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 A 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)

^*Corresponding author for this work

Medical Sciences

Research output: Contribution to journal › Article › peer-review

34 Citations (Scopus)

10 Downloads (Pure)

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.

Original language	English
Pages (from-to)	381-391
Number of pages	11
Journal	The Journal of pathology
Volume	247
Issue number	3
Early online date	25 Jan 2019
DOIs	https://doi.org/10.1002/path.5197
Publication status	Published - 1 Mar 2019

Bibliographical 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

Keywords

DOCK4
bone metastasis
breast cancer
biomarker
proteomics

UN SDGs

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

Access to Document

10.1002/path.5197Licence: CC BY

Accepted Manuscript
This is the peer reviewed version of the following article: Westbrook, J. A., Wood, S. L., Cairns, D. A., McMahon, K. , Gahlaut, R. , Thygesen, H. , Shires, M. , Roberts, S. , Marshall, H. , Oliva, M. R., Dunning, M. J., Hanby, A. M., Selby, P. J., Speirs, V. , Mavria, G. , Coleman, R. E. and Brown, J. E. (2019), Identification and validation of DOCK4 as a potential biomarker for risk of bone metastasis development in patients with early breast cancer. J. Pathol., 247: 381-391., which has been published in final form at https://doi.org/10.1002/path.5197. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.
Accepted author manuscript, 2.07 MBLicence: Unspecified
Westbrook_et_al_JofPat_IdentificationAndValidationOfDOCK4_VOR
This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. https://creativecommons.org/licenses/by/4.0/
Final published version, 2 MBLicence: CC BY

Cite this

Westbrook, J. A., Wood, S. L., Cairns, D. A., McMahon, K., Gahlaut, R., Thygesen, H., Shires, M., Roberts, S., Marshall, H., Oliva, M. R., Dunning, M. J., Hanby, A. M., Selby, P. J., Speirs, V., Mavria, G., Coleman, R. E., & Brown, J. E. (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, 247(3), 381-391. https://doi.org/10.1002/path.5197

Westbrook, JA, Wood, SL, Cairns, DA, 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

@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.",

keywords = "DOCK4, bone metastasis, breast cancer, biomarker, proteomics",

author = "Westbrook, {Jules A} and Wood, {Steven L} and Cairns, {David A} 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 = mar,

day = "1",

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 A

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 - 2019/3/1

Y1 - 2019/3/1

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.

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.

KW - DOCK4

KW - bone metastasis

KW - breast cancer

KW - biomarker

KW - proteomics

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

C2 - 30426503

SN - 0022-3417

VL - 247

SP - 381

EP - 391

JO - The Journal of pathology

JF - The Journal of pathology

IS - 3

ER -

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

Abstract

Bibliographical note

Keywords

UN SDGs

Access to Document

Other files and links

Fingerprint

Valerie Speirs

Cite this

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

Abstract

Bibliographical note

Keywords

UN SDGs

Access to Document

Other files and links

Fingerprint

Profiles

Valerie Speirs

Cite this