Show simple item record

dc.contributor.authorO´Leary, Patrick C
dc.contributor.authorPenny, Sarah A
dc.contributor.authorDolan, Roisin T
dc.contributor.authorKelly, Catherine M
dc.contributor.authorMadden, Stephen F
dc.contributor.authorRexhepaj, Elton
dc.contributor.authorBrennan, Donal J
dc.contributor.authorMcCann, Amanda H
dc.contributor.authorPontén, Fredrik
dc.contributor.authorUhlén, Mathias
dc.contributor.authorZagozdzon, Radoslaw
dc.contributor.authorDuffy, Michael J
dc.contributor.authorKell, Malcolm R
dc.contributor.authorJirström, Karin
dc.contributor.authorGallagher, William M
dc.date.accessioned2013-06-24T18:13:34Z
dc.date.available2013-06-24T18:13:34Z
dc.date.issued2013-04-02
dc.identifier.citationBMC Cancer. 2013 Apr 02;13(1):175
dc.identifier.urihttp://dx.doi.org/10.1186/1471-2407-13-175
dc.identifier.urihttp://hdl.handle.net/10147/294476
dc.description.abstractAbstract Background Although omic-based discovery approaches can provide powerful tools for biomarker identification, several reservations have been raised regarding the clinical applicability of gene expression studies, such as their prohibitive cost. However, the limited availability of antibodies is a key barrier to the development of a lower cost alternative, namely a discrete collection of immunohistochemistry (IHC)-based biomarkers. The aim of this study was to use a systematic approach to generate and screen affinity-purified, mono-specific antibodies targeting progression-related biomarkers, with a view towards developing a clinically applicable IHC-based prognostic biomarker panel for breast cancer. Methods We examined both in-house and publicly available breast cancer DNA microarray datasets relating to invasion and metastasis, thus identifying a cohort of candidate progression-associated biomarkers. Of these, 18 antibodies were released for extended analysis. Validated antibodies were screened against a tissue microarray (TMA) constructed from a cohort of consecutive breast cancer cases (n = 512) to test the immunohistochemical surrogate signature. Results Antibody screening revealed 3 candidate prognostic markers: the cell cycle regulator, Anillin (ANLN); the mitogen-activated protein kinase, PDZ-Binding Kinase (PBK); and the estrogen response gene, PDZ-Domain Containing 1 (PDZK1). Increased expression of ANLN and PBK was associated with poor prognosis, whilst increased expression of PDZK1 was associated with good prognosis. A 3-marker signature comprised of high PBK, high ANLN and low PDZK1 expression was associated with decreased recurrence-free survival (p < 0.001) and breast cancer-specific survival (BCSS) (p < 0.001). This novel signature was associated with high tumour grade (p < 0.001), positive nodal status (p = 0.029), ER-negativity (p = 0.006), Her2-positivity (p = 0.036) and high Ki67 status (p < 0.001). However, multivariate Cox regression demonstrated that the signature was not a significant predictor of BCSS (HR = 6.38; 95% CI = 0.79-51.26, p = 0.082). Conclusions We have developed a comprehensive biomarker pathway that extends from discovery through to validation on a TMA platform. This proof-of-concept study has resulted in the identification of a novel 3-protein prognostic panel. Additional biochemical markers, interrogated using this high-throughput platform, may further augment the prognostic accuracy of this panel to a point that may allow implementation into routine clinical practice.
dc.titleSystematic antibody generation and validation via tissue microarray technology leading to identification of a novel protein prognostic panel in breast cancer
dc.typeJournal Article
dc.language.rfc3066en
dc.rights.holderPatrick C O´Leary et al.; licensee BioMed Central Ltd.
dc.description.statusPeer Reviewed
dc.date.updated2013-05-30T15:07:12Z
refterms.dateFOA2018-08-23T05:49:01Z
html.description.abstractAbstract Background Although omic-based discovery approaches can provide powerful tools for biomarker identification, several reservations have been raised regarding the clinical applicability of gene expression studies, such as their prohibitive cost. However, the limited availability of antibodies is a key barrier to the development of a lower cost alternative, namely a discrete collection of immunohistochemistry (IHC)-based biomarkers. The aim of this study was to use a systematic approach to generate and screen affinity-purified, mono-specific antibodies targeting progression-related biomarkers, with a view towards developing a clinically applicable IHC-based prognostic biomarker panel for breast cancer. Methods We examined both in-house and publicly available breast cancer DNA microarray datasets relating to invasion and metastasis, thus identifying a cohort of candidate progression-associated biomarkers. Of these, 18 antibodies were released for extended analysis. Validated antibodies were screened against a tissue microarray (TMA) constructed from a cohort of consecutive breast cancer cases (n&#8201;=&#8201;512) to test the immunohistochemical surrogate signature. Results Antibody screening revealed 3 candidate prognostic markers: the cell cycle regulator, Anillin (ANLN); the mitogen-activated protein kinase, PDZ-Binding Kinase (PBK); and the estrogen response gene, PDZ-Domain Containing 1 (PDZK1). Increased expression of ANLN and PBK was associated with poor prognosis, whilst increased expression of PDZK1 was associated with good prognosis. A 3-marker signature comprised of high PBK, high ANLN and low PDZK1 expression was associated with decreased recurrence-free survival (p&#8201;&lt;&#8201;0.001) and breast cancer-specific survival (BCSS) (p&#8201;&lt;&#8201;0.001). This novel signature was associated with high tumour grade (p&#8201;&lt;&#8201;0.001), positive nodal status (p&#8201;=&#8201;0.029), ER-negativity (p&#8201;=&#8201;0.006), Her2-positivity (p&#8201;=&#8201;0.036) and high Ki67 status (p&#8201;&lt;&#8201;0.001). However, multivariate Cox regression demonstrated that the signature was not a significant predictor of BCSS (HR&#8201;=&#8201;6.38; 95% CI&#8201;=&#8201;0.79-51.26, p&#8201;=&#8201;0.082). Conclusions We have developed a comprehensive biomarker pathway that extends from discovery through to validation on a TMA platform. This proof-of-concept study has resulted in the identification of a novel 3-protein prognostic panel. Additional biochemical markers, interrogated using this high-throughput platform, may further augment the prognostic accuracy of this panel to a point that may allow implementation into routine clinical practice.


Files in this item

Thumbnail
Name:
1471-2407-13-175.xml
Size:
121.9Kb
Format:
XML
Thumbnail
Name:
1471-2407-13-175.pdf
Size:
3.139Mb
Format:
PDF
Thumbnail
Name:
1471-2407-13-175-S1.PDF
Size:
370.0Kb
Format:
PDF
Thumbnail
Name:
1471-2407-13-175-S2.JPEG
Size:
1.021Mb
Format:
JPEG image

This item appears in the following Collection(s)

Show simple item record