Highly sensitive proximity mediated immunoassay reveals HER2 status conversion in the circulating tumor cells of metastatic breast cancer patients

<p>Abstract</p> <p>Background</p> <p>The clinical benefits associated with targeted oncology agents are generally limited to subsets of patients. Even with favorable biomarker profiles, many patients do not respond or acquire resistance. Existing technologies are ineffe...

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Main Authors: Kim Phillip, Liu Xinjun, Lee Tani, Liu Limin, Barham Robert, Kirkland Richard, Leesman Glen, Kuller Anne, Ybarrondo Belen, Ng Shi-Chung, Singh Sharat
Format: Article
Language:English
Published: BMC 2011-12-01
Series:Proteome Science
Subjects:
Online Access:http://www.proteomesci.com/content/9/1/75
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spelling doaj-680021ef8a544097974e08b65fdc82372020-11-24T22:18:46ZengBMCProteome Science1477-59562011-12-01917510.1186/1477-5956-9-75Highly sensitive proximity mediated immunoassay reveals HER2 status conversion in the circulating tumor cells of metastatic breast cancer patientsKim PhillipLiu XinjunLee TaniLiu LiminBarham RobertKirkland RichardLeesman GlenKuller AnneYbarrondo BelenNg Shi-ChungSingh Sharat<p>Abstract</p> <p>Background</p> <p>The clinical benefits associated with targeted oncology agents are generally limited to subsets of patients. Even with favorable biomarker profiles, many patients do not respond or acquire resistance. Existing technologies are ineffective for treatment monitoring as they provide only static and limited information and require substantial amounts of tissue. Therefore, there is an urgent need to develop methods that can profile potential therapeutic targets with limited clinical specimens during the course of treatment.</p> <p>Methods</p> <p>We have developed a novel proteomics-based assay, Collaborative Enzyme Enhanced Reactive-immunoassay (CEER) that can be used for analyzing clinical samples. CEER utilizes the formation of unique immuno-complex between capture-antibodies and two additional detector-Abs on a microarray surface. One of the detector-Abs is conjugated to glucose oxidase (GO), and the other is conjugated to Horse Radish Peroxidase (HRP). Target detection requires the presence of both detector-Abs because the enzyme channeling event between GO and HRP will not occur unless both Abs are in close proximity.</p> <p>Results</p> <p>CEER was able to detect single-cell level expression and phosphorylation of human epidermal growth factor receptor 2 (HER2) and human epidermal growth factor receptor 1 (HER1) in breast cancer (BCa) systems. The shift in phosphorylation profiles of receptor tyrosine kinases (RTKs) and other signal transduction proteins upon differential ligand stimulation further demonstrated extreme assay specificity in a multiplexed array format. HER2 analysis by CEER in 227 BCa tissues showed superior accuracy when compared to the outcome from immunohistochemistry (IHC) (83% vs. 96%). A significant incidence of HER2 status alteration with recurrent disease was observed via circulating tumor cell (CTC) analysis, suggesting an evolving and dynamic disease progression. HER2-positive CTCs were found in 41% (7/17) while CTCs with significant HER2-activation without apparent over-expression were found in 18% (3/17) of relapsed BCa patients with HER2-negative primary tumors. The apparent 'HER2 status conversion' observed in recurrent BCa may have significant implications on understanding breast cancer metastasis and associated therapeutic development.</p> <p>Conclusion</p> <p>CEER can be multiplexed to analyze pathway proteins in a comprehensive manner with extreme specificity and sensitivity. This format is ideal for analyzing clinical samples with limited availability.</p> http://www.proteomesci.com/content/9/1/75Companion diagnosticsCollaborative enzyme enhanced reactive-immunoassayMetastatic breast cancer, Circulating tumor cellsHER2 conversion
collection DOAJ
language English
format Article
sources DOAJ
author Kim Phillip
Liu Xinjun
Lee Tani
Liu Limin
Barham Robert
Kirkland Richard
Leesman Glen
Kuller Anne
Ybarrondo Belen
Ng Shi-Chung
Singh Sharat
spellingShingle Kim Phillip
Liu Xinjun
Lee Tani
Liu Limin
Barham Robert
Kirkland Richard
Leesman Glen
Kuller Anne
Ybarrondo Belen
Ng Shi-Chung
Singh Sharat
Highly sensitive proximity mediated immunoassay reveals HER2 status conversion in the circulating tumor cells of metastatic breast cancer patients
Proteome Science
Companion diagnostics
Collaborative enzyme enhanced reactive-immunoassay
Metastatic breast cancer, Circulating tumor cells
HER2 conversion
author_facet Kim Phillip
Liu Xinjun
Lee Tani
Liu Limin
Barham Robert
Kirkland Richard
Leesman Glen
Kuller Anne
Ybarrondo Belen
Ng Shi-Chung
Singh Sharat
author_sort Kim Phillip
title Highly sensitive proximity mediated immunoassay reveals HER2 status conversion in the circulating tumor cells of metastatic breast cancer patients
title_short Highly sensitive proximity mediated immunoassay reveals HER2 status conversion in the circulating tumor cells of metastatic breast cancer patients
title_full Highly sensitive proximity mediated immunoassay reveals HER2 status conversion in the circulating tumor cells of metastatic breast cancer patients
title_fullStr Highly sensitive proximity mediated immunoassay reveals HER2 status conversion in the circulating tumor cells of metastatic breast cancer patients
title_full_unstemmed Highly sensitive proximity mediated immunoassay reveals HER2 status conversion in the circulating tumor cells of metastatic breast cancer patients
title_sort highly sensitive proximity mediated immunoassay reveals her2 status conversion in the circulating tumor cells of metastatic breast cancer patients
publisher BMC
series Proteome Science
issn 1477-5956
publishDate 2011-12-01
description <p>Abstract</p> <p>Background</p> <p>The clinical benefits associated with targeted oncology agents are generally limited to subsets of patients. Even with favorable biomarker profiles, many patients do not respond or acquire resistance. Existing technologies are ineffective for treatment monitoring as they provide only static and limited information and require substantial amounts of tissue. Therefore, there is an urgent need to develop methods that can profile potential therapeutic targets with limited clinical specimens during the course of treatment.</p> <p>Methods</p> <p>We have developed a novel proteomics-based assay, Collaborative Enzyme Enhanced Reactive-immunoassay (CEER) that can be used for analyzing clinical samples. CEER utilizes the formation of unique immuno-complex between capture-antibodies and two additional detector-Abs on a microarray surface. One of the detector-Abs is conjugated to glucose oxidase (GO), and the other is conjugated to Horse Radish Peroxidase (HRP). Target detection requires the presence of both detector-Abs because the enzyme channeling event between GO and HRP will not occur unless both Abs are in close proximity.</p> <p>Results</p> <p>CEER was able to detect single-cell level expression and phosphorylation of human epidermal growth factor receptor 2 (HER2) and human epidermal growth factor receptor 1 (HER1) in breast cancer (BCa) systems. The shift in phosphorylation profiles of receptor tyrosine kinases (RTKs) and other signal transduction proteins upon differential ligand stimulation further demonstrated extreme assay specificity in a multiplexed array format. HER2 analysis by CEER in 227 BCa tissues showed superior accuracy when compared to the outcome from immunohistochemistry (IHC) (83% vs. 96%). A significant incidence of HER2 status alteration with recurrent disease was observed via circulating tumor cell (CTC) analysis, suggesting an evolving and dynamic disease progression. HER2-positive CTCs were found in 41% (7/17) while CTCs with significant HER2-activation without apparent over-expression were found in 18% (3/17) of relapsed BCa patients with HER2-negative primary tumors. The apparent 'HER2 status conversion' observed in recurrent BCa may have significant implications on understanding breast cancer metastasis and associated therapeutic development.</p> <p>Conclusion</p> <p>CEER can be multiplexed to analyze pathway proteins in a comprehensive manner with extreme specificity and sensitivity. This format is ideal for analyzing clinical samples with limited availability.</p>
topic Companion diagnostics
Collaborative enzyme enhanced reactive-immunoassay
Metastatic breast cancer, Circulating tumor cells
HER2 conversion
url http://www.proteomesci.com/content/9/1/75
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