Antibody-based detection of ERG rearrangement-positive prostate cancer.
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Journal: 2010/November - Neoplasia
ISSN: 1476-5586
PUBMED: 20651988
Abstract:
TMPRSS2-ERG gene fusions occur in 50% of prostate cancers and result in the overexpression of a chimeric fusion transcript that encodes a truncated ERG product. Previous attempts to detect truncated ERG products have been hindered by a lack of specific antibodies. Here, we characterize a rabbit anti-ERG monoclonal antibody (clone EPR 3864; Epitomics, Burlingame, CA) using immunoblot analysis on prostate cancer cell lines, synthetic TMPRSS2-ERG constructs, chromatin immunoprecipitation, and immunofluorescence. We correlated ERG protein expression with the presence of ERG gene rearrangements in prostate cancer tissues using a combined immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) analysis. We independently evaluated two patient cohorts and observed ERG expression confined to prostate cancer cells and high-grade prostatic intraepithelial neoplasia associated with ERG-positive cancer, as well as vessels and lymphocytes (where ERG has a known biologic role). Image analysis of 131 cases demonstrated nearly 100% sensitivity for detecting ERG rearrangement prostate cancer, with only 2 (1.5%) of 131 cases demonstrating strong ERG protein expression without any known ERG gene fusion. The combined pathology evaluation of 207 patient tumors for ERG protein expression had 95.7% sensitivity and 96.5% specificity for determining ERG rearrangement prostate cancer. In conclusion, this study qualifies a specific anti-ERG antibody and demonstrates exquisite association between ERG gene rearrangement and truncated ERG protein product expression. Given the ease of performing IHC versus FISH, ERG protein expression may be useful for molecularly subtyping prostate cancer based on ERG rearrangement status and suggests clinical utility in prostate needle biopsy evaluation.
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Neoplasia 12(7): 590-598
PMID: 20651988

Antibody-Based Detection of <em>ERG</em> Rearrangement-Positive Prostate Cancer<sup><a href="#FN1" rid="FN1" class=" fn">1</a>,</sup><sup><a href="#FN2" rid="FN2" class=" fn">2</a></sup>

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Supplementary Figures and Tables:
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Department of Pathology and Laboratory Medicine, Weill Cornell Medical College, New York, NY, USA
Michigan Center for Translational Pathology, Ann Arbor, MI, USA
Department of Pathology, University of Michigan, Ann Arbor, MI, USA
Division of Biostatistics and Epidemiology, Department of Public Health, Weill Cornell Medical College, New York, NY, USA
Department of Urology, Weill Cornell Medical College, New York, NY, USA
Ventana, a member of the Roche Group, Tucson, AZ, USA
Institute for Computational Biomedicine, Weill Cornell Medical College, New York, NY, USA
Howard Hughes Medical Institute, Chevy Chase, MD, USA
Department of Urology, University of Michigan, Ann Arbor, MI, USA
Comprehensive Cancer Center, University of Michigan, Ann Arbor, MI, USA
These authors equally contributed to this work.
These authors share senior authorship.
Address all correspondence to: Mark A. Rubin, MD, Department of Pathology and Laboratory Medicine, 1300 York Ave, Room C 410-A, New York, NY 10065. E-mail: ude.llenroc.dem@amnibur
Address all correspondence to: Mark A. Rubin, MD, Department of Pathology and Laboratory Medicine, 1300 York Ave, Room C 410-A, New York, NY 10065. E-mail: ude.llenroc.dem@amnibur
Received 2010 May 23; Revised 2010 Jun 23; Accepted 2010 Jun 23.
Copyright © 2010 Neoplasia Press, Inc. All rights reserved

Abstract

TMPRSS2-ERG gene fusions occur in 50% of prostate cancers and result in the overexpression of a chimeric fusion transcript that encodes a truncated ERG product. Previous attempts to detect truncated ERG products have been hindered by a lack of specific antibodies. Here, we characterize a rabbit anti-ERG monoclonal antibody (clone EPR 3864; Epitomics, Burlingame, CA) using immunoblot analysis on prostate cancer cell lines, synthetic TMPRSS2-ERG constructs, chromatin immunoprecipitation, and immunofluorescence. We correlated ERG protein expression with the presence of ERG gene rearrangements in prostate cancer tissues using a combined immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) analysis. We independently evaluated two patient cohorts and observed ERG expression confined to prostate cancer cells and high-grade prostatic intraepithelial neoplasia associated with ERG-positive cancer, as well as vessels and lymphocytes (where ERG has a known biologic role). Image analysis of 131 cases demonstrated nearly 100% sensitivity for detecting ERG rearrangement prostate cancer, with only 2 (1.5%) of 131 cases demonstrating strong ERG protein expression without any known ERG gene fusion. The combined pathology evaluation of 207 patient tumors for ERG protein expression had 95.7% sensitivity and 96.5% specificity for determining ERG rearrangement prostate cancer. In conclusion, this study qualifies a specific anti-ERG antibody and demonstrates exquisite association between ERG gene rearrangement and truncated ERG protein product expression. Given the ease of performing IHC versus FISH, ERG protein expression may be useful for molecularly subtyping prostate cancer based on ERG rearrangement status and suggests clinical utility in prostate needle biopsy evaluation.

Abstract

Acknowledgments

The authors thank Robert Kim of the WCMC Translational Research Program of the Department of Pathology and Laboratory Medicine and Yifang Liu for technical expertise and Ashley Santa Cruz at Ventana/Roche for optimizing the final protocol for ERG IHC. The authors also thank Javed Siddiqui of the UM Prostate SPORE Tissue/Informatics Core and Nalla Palanisamy for managing previous FISH probe preparations at UM.

Acknowledgments

Abbreviations

ERGv-ets erythroblastosis virus E26 oncogene homolog (avian)
TMAtissue microarray
FISHfluorescence in situ hybridization
ChIPchromatin immunoprecipitation
Abbreviations

Footnotes

This study was supported by National Cancer Institute (NCI) grants CA125612 (F.D. and M.A.R.), U01CA111275-06 Early Detection Research Network (S.A.T., M.A.R., and A.M.C.), and R01CA132874 (A.M.C.) and Prostate SPORE P50CA69568 (A.M.C. to M.A.R.). S.A.T. is supported by a Young Investigator Award from the Prostate Cancer Foundation. A.M.C. is supported by the Doris Duke Charitable Foundation Clinical Scientist Award, Burroughs Welcome Foundation Award in Clinical Translational Research, and the Prostate Cancer Foundation. A.M.C. is an American Cancer Society Research Professor. F.D. is supported by a Developmental Project Award from Dana-Farber Harvard Cancer Center Prostate Cancer SPORE.

This article refers to supplementary materials, which are designated by Figures W1 to W5 and are available online at www.neoplasia.com.

Footnotes

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