Journal of Clinical Oncology, Vol 19, Issue 10
(May), 2001: 2714-2721
© 2001 American Society for Clinical Oncology
Discrepancies in Clinical Laboratory Testing of Eligibility for Trastuzumab Therapy: Apparent Immunohistochemical False-Positives Do Not Get the Message
By R.R. Tubbs,
J.D. Pettay,
P.C. Roche,
M.H. Stoler,
R.B. Jenkins,
T.M. Grogan
From the Department of Clinical Pathology, Cleveland Clinic Foundation, Cleveland, OH; Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN; Robert E. Fechner Laboratory of Surgical Pathology, University of Virginia Health System, Richmond, VA; and Department of Pathology, University of Arizona School of Medicine, Tucson, AZ.
Address reprint requests to Raymond R. Tubbs, DO, Cleveland Clinic Foundation, 9500 Euclid Ave, L11, Cleveland, OH 44195; email: TubbsR{at}ccf.org
BACKGROUND: Several studies have reported what seem to be false-positive results using the Food and Drug Administration (FDA)–approved HercepTest (Dako Corp, Carpinteria, CA) to profile Her-2/neu amplification and overproduction in breast carcinoma. False-positive status has been based on comparisons with gene copy enumeration by fluorescence in situ hybridization (FISH) and with comparisons to immunohistochemistry (IMH) results using a monoclonal antibody. However, simple overexpression by tumor cells that have normal gene copy has not been evaluated by profiling mRNA expression, ie, such cases could simply represent true-positive, transcriptionally upregulated overexpression.
MATERIALS AND METHODS: Four hundred infiltrating ductal carcinomas of breast were evaluated by IMH using monoclonal (CB11; Ventana Medical Systems, Inc, Tucson, AZ) and polyclonal (HercepTest; Dako) antibodies after antigen retrieval (AR). A polyclonal antibody sans AR (PCA/SAR) was also used. All IMH stains were evaluated and scored according to the guidelines for the FDA-approved HercepTest. A total of 145 of 400 carcinomas were subsequently evaluated by direct and digoxigenin-labeled (Dig) FISH, and 144 of 400 were evaluated by detection of mRNA overexpression via autoradiographic RNA:RNA in situ hybridization.
RESULTS: Overall HercepTest/CB11 IMH discordance was 12%. Expression of mRNA was highly concordant with FISH and DigFISH amplification and with CB11 and PCA/SAR immunohistology. IMH false-positive cases (no Her-2/neu gene amplification) occurred with both HercepTest (23%) and CB11 (17%), and the majority of false-positive results (34 of 44) were scored as 2+. All 2+ false-positive cases were mRNA-negative. Combined results of HercepTest and CB11 showed that 79% (38 of 48) of 3+ cases were Her-2/neu gene amplified, but only 17% (seven of 41) of 2+ cases had increased gene copy.
CONCLUSION: Discordant HercepTest/FISH results, and to a lesser extent discordance with CB11 IMH, are most commonly false-positive results with a score of 2+. The 2+ score as defined in the guidelines for the FDA-approved HercepTest should not be used as a criterion for trastuzumab therapy unless confirmed by FISH. Determination of Her-2 gene copy number by FISH may be a more accurate and reliable method for selecting patients eligible for trastuzumab therapy.
T.M.G. is also a consultant and director of Ventana Medical Systems, Inc, Tucson, AZ.
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Am J Clin Pathol,
July 1, 2009;
132(1):
133 - 138.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J Zustin, K Boddin, M C Tsourlakis, E Burandt, M Mirlacher, F Jaenicke, J Izbicki, W Ruether, J M Rueger, C Bokemeyer, et al.
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J. Clin. Pathol.,
June 1, 2009;
62(6):
542 - 546.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. S. Ross, E. A. Slodkowska, W. F. Symmans, L. Pusztai, P. M. Ravdin, and G. N. Hortobagyi
The HER-2 Receptor and Breast Cancer: Ten Years of Targeted Anti-HER-2 Therapy and Personalized Medicine
Oncologist,
April 1, 2009;
14(4):
320 - 368.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
G. Sauter, J. Lee, J. M.S. Bartlett, D. J. Slamon, and M. F. Press
Guidelines for Human Epidermal Growth Factor Receptor 2 Testing: Biologic and Methodologic Considerations
J. Clin. Oncol.,
March 10, 2009;
27(8):
1323 - 1333.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. A. Iverson, C. Gillett, P. Cane, C. D. Santini, T. M. Vess, L. Kam-Morgan, A. Wang, M. Eisenberg, C. M. Rowland, J. J. Hessling, et al.
A Single-Tube Quantitative Assay for mRNA Levels of Hormonal and Growth Factor Receptors in Breast Cancer Specimens
J. Mol. Diagn.,
March 1, 2009;
11(2):
117 - 130.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S Di Palma, N Collins, M Bilous, A Sapino, M Mottolese, N Kapranos, F Schmitt, and J Isola
A quality assurance exercise to evaluate the accuracy and reproducibility of chromogenic in situ hybridisation for HER2 analysis in breast cancer
J. Clin. Pathol.,
June 1, 2008;
61(6):
757 - 760.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. C. Wolff, M. E. H. Hammond, J. N. Schwartz, D. F. Hayes, and On behalf of the ASCO/CAP Panel on HER-2 Testing i
In Reply
J. Clin. Oncol.,
September 1, 2007;
25(25):
4021 - 4023.
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. L. Costantini, C. Chan, Z. Cai, K. A. Vallis, and R. M. Reilly
111In-Labeled Trastuzumab (Herceptin) Modified with Nuclear Localization Sequences (NLS): An Auger Electron-Emitting Radiotherapeutic Agent for HER2/neu-Amplified Breast Cancer
J. Nucl. Med.,
August 1, 2007;
48(8):
1357 - 1368.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. Barrett, H. Magee, D. O'Toole, S. Daly, and M. Jeffers
Amplification of the HER2 gene in breast cancers testing 2+ weak positive by HercepTest immunohistochemistry: false-positive or false-negative immunohistochemistry?
J. Clin. Pathol.,
June 1, 2007;
60(6):
690 - 693.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. S. Ross, W. F. Symmans, L. Pusztai, and G. N. Hortobagyi
Standardizing Slide-Based Assays in Breast Cancer: Hormone Receptors, HER2, and Sentinel Lymph Nodes
Clin. Cancer Res.,
May 15, 2007;
13(10):
2831 - 2835.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Glazyrin, X. Shen, V. Blanc, and J. F. Eliason
Direct Detection of Herceptin/Trastuzumab Binding on Breast Tissue Sections
J. Histochem. Cytochem.,
January 1, 2007;
55(1):
25 - 33.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. C. Wolff, M. E. H. Hammond, J. N. Schwartz, K. L. Hagerty, D. C. Allred, R. J. Cote, M. Dowsett, P. L. Fitzgibbons, W. M. Hanna, A. Langer, et al.
American Society of Clinical Oncology/College of American Pathologists Guideline Recommendations for Human Epidermal Growth Factor Receptor 2 Testing in Breast Cancer
J. Clin. Oncol.,
January 1, 2007;
25(1):
118 - 145.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. Dal Lago, V. Durbecq, C. Desmedt, R. Salgado, T. Verjat, L. Lespagnard, Y. Ma, I. Veys, A. Di Leo, C. Sotiriou, et al.
Correction for chromosome-17 is critical for the determination of true Her-2/neu gene amplification status in breast cancer.
Mol. Cancer Ther.,
October 1, 2006;
5(10):
2572 - 2579.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
E. Li-Ning-T, R. Ronchetti, C. Torres-Cabala, and M. J. Merino
Role of Chromogenic in Situ Hybridization (CISHTM) in the Evaluation of HER2 Status in Breast Carcinoma: Comparison with Immunohistochemistry and Fish
International Journal of Surgical Pathology,
October 1, 2005;
13(4):
343 - 351.
[Abstract]
[PDF]
|
|
|
|
|
|
|
M. F. Press, G. Sauter, L. Bernstein, I. E. Villalobos, M. Mirlacher, J.-Y. Zhou, R. Wardeh, Y.-T. Li, R. Guzman, Y. Ma, et al.
Diagnostic Evaluation of HER-2 as a Molecular Target: An Assessment of Accuracy and Reproducibility of Laboratory Testing in Large, Prospective, Randomized Clinical Trials
Clin. Cancer Res.,
September 15, 2005;
11(18):
6598 - 6607.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C M Ellis, M J Dyson, T J Stephenson, and E L Maltby
HER2 amplification status in breast cancer: a comparison between immunohistochemical staining and fluorescence in situ hybridisation using manual and automated quantitative image analysis scoring techniques
J. Clin. Pathol.,
July 1, 2005;
58(7):
710 - 714.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H. Yaziji, T. S. Barry, and A. M. Gown
HER-2 Testing in Breast Cancer--Reply
JAMA,
October 20, 2004;
292(15):
1818 - 1818.
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. Zhu, L. W. C. Chow, W. T. Y. Loo, X.-Y. Guan, and M. Toi
Her2/neu Expression Predicts the Response to Antiaromatase Neoadjuvant Therapy in Primary Breast Cancer: Subgroup Analysis from Celecoxib Antiaromatase Neoadjuvant Trial
Clin. Cancer Res.,
July 15, 2004;
10(14):
4639 - 4644.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. Isola, M. Tanner, A. Forsyth, T. G. Cooke, A. D. Watters, and J. M. S. Bartlett
Interlaboratory Comparison of HER-2 Oncogene Amplification as Detected by Chromogenic and Fluorescence in situ Hybridization
Clin. Cancer Res.,
July 15, 2004;
10(14):
4793 - 4798.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
C. T. Dang, A. J. Dannenberg, K. Subbaramaiah, M. N. Dickler, M. M. Moasser, A. D. Seidman, G. M. D'Andrea, M. Theodoulou, K. S. Panageas, L. Norton, et al.
Phase II Study of Celecoxib and Trastuzumab in Metastatic Breast Cancer Patients Who Have Progressed after Prior Trastuzumab-Based Treatments
Clin. Cancer Res.,
June 15, 2004;
10(12):
4062 - 4067.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
H. Yaziji, L. C. Goldstein, T. S. Barry, R. Werling, H. Hwang, G. K. Ellis, J. R. Gralow, R. B. Livingston, and A. M. Gown
HER-2 Testing in Breast Cancer Using Parallel Tissue-Based Methods
JAMA,
April 28, 2004;
291(16):
1972 - 1977.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
E. L. Wiley and L. K. Diaz
High-Quality HER-2 Testing: Setting a Standard for Oncologic Biomarker Assessment
JAMA,
April 28, 2004;
291(16):
2019 - 2020.
[Full Text]
[PDF]
|
|
|
|
|
|
|
L. K. Diaz, R. Gupta, N. Kidwai, N. Sneige, and E. L. Wiley
The Use of TMA for Interlaboratory Validation of FISH Testing for Detection of HER2 Gene Amplification in Breast Cancer
J. Histochem. Cytochem.,
April 1, 2004;
52(4):
501 - 507.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. S. Ross, J. A. Fletcher, K. J. Bloom, G. P. Linette, J. Stec, W. F. Symmans, L. Pusztai, and G. N. Hortobagyi
Targeted Therapy in Breast Cancer: The HER-2/neu Gene and Protein
Mol. Cell. Proteomics,
April 1, 2004;
3(4):
379 - 398.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
E. B. Elkin, M. C. Weinstein, E. P. Winer, K. M. Kuntz, S. J. Schnitt, and J. C. Weeks
HER-2 Testing and Trastuzumab Therapy for Metastatic Breast Cancer: A Cost-Effectiveness Analysis
J. Clin. Oncol.,
March 1, 2004;
22(5):
854 - 863.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. Lal, P. A. Salazar, M. Ladanyi, and B. Chen
Impact of Polysomy 17 on HER-2/neu Immunohistochemistry in Breast Carcinomas without HER-2/neu Gene Amplification
J. Mol. Diagn.,
August 1, 2003;
5(3):
155 - 159.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
J. S. Ross, J. A. Fletcher, G. P. Linette, J. Stec, E. Clark, M. Ayers, W. F. Symmans, L. Pusztai, and K. J. Bloom
The HER-2/neu Gene and Protein in Breast Cancer 2003: Biomarker and Target of Therapy
Oncologist,
August 1, 2003;
8(4):
307 - 325.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
M. Varella-Garcia
Molecular Cytogenetics in Solid Tumors: Laboratorial Tool for Diagnosis, Prognosis, and Therapy
Oncologist,
February 1, 2003;
8(1):
45 - 58.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. L. Lark, C. A. Livasy, B. Calvo, L. Caskey, D. T. Moore, X. Yang, and W. G. Cance
Overexpression of Focal Adhesion Kinase in Primary Colorectal Carcinomas and Colorectal Liver Metastases: Immunohistochemistry and Real-Time PCR Analyses
Clin. Cancer Res.,
January 1, 2003;
9(1):
215 - 222.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. Birner, G. Oberhuber, M. F. Press, L. Bernstein, K. Flom, and D. Slamon
Is Fluorescence In Situ Hybridization Really Superior to HercepTest?
J. Clin. Oncol.,
December 1, 2002;
20(23):
4607 - 4609.
[Full Text]
[PDF]
|
|
|
|
|
|
|
S Soule, L Baldridge, K Kirkpatrick, L Cheng, J L Gilbert, L R Smith, V C Thurston, G H Vance, L Einhorn, and K Miller
HER-2/neu expression in germ cell tumours
J. Clin. Pathol.,
September 1, 2002;
55(9):
656 - 658.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
P. S. Bernard and C. T. Wittwer
Real-Time PCR Technology for Cancer Diagnostics
Clin. Chem.,
August 1, 2002;
48(8):
1178 - 1185.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
S. Paik, J. Bryant, E. Tan-Chiu, E. Romond, W. Hiller, K. Park, A. Brown, G. Yothers, S. Anderson, R. Smith, et al.
Real-World Performance of HER2 Testing--National Surgical Adjuvant Breast and Bowel Project Experience
J Natl Cancer Inst,
June 5, 2002;
94(11):
852 - 854.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
R. Tubbs, J. Pettay, M. Skacel, R. Powell, M. Stoler, P. Roche, and J. Hainfeld
Gold-Facilitated in Situ Hybridization : A Bright-Field Autometallographic Alternative to Fluorescence in Situ Hybridization for Detection of HER-2/neu Gene Amplification
Am. J. Pathol.,
May 1, 2002;
160(5):
1589 - 1595.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
D. G. Thomas, T. J. Giordano, D. Sanders, J. S. Biermann, and L. Baker
Absence of HER2/neu Gene Expression in Osteosarcoma and Skeletal Ewing's Sarcoma
Clin. Cancer Res.,
March 1, 2002;
8(3):
788 - 793.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
E. A. Perez, P. C. Roche, R. B. Jenkins, C. A. Reynolds, K. C. Halling, J. N. Ingle, and L. E. Wold
HER2 Testing in Patients With Breast Cancer: Poor Correlation Between Weak Positivity by Immunohistochemistry and Gene Amplification by Fluorescence In Situ Hybridization
Mayo Clin. Proc.,
February 1, 2002;
77(2):
148 - 154.
[Abstract]
[PDF]
|
|
|
|
|
|
|
D. W. Nichols, D. J. Wolff, S. Self, J. S. Metcalf, D. Jacobs, R. Kneuper-Hall, and J. C. Cate IV
A Testing Algorithm for Determination of HER2 Status in Patients with Breast Cancer
Ann. Clin. Lab. Sci.,
January 1, 2002;
32(1):
3 - 11.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
W. K. Funkhouser and K. Kaiser-Rogers
Significance of, and Optimal Screening for, HER-2 Gene Amplification and Protein Overexpression in Breast Carcinoma
Ann. Clin. Lab. Sci.,
October 1, 2001;
31(4):
349 - 358.
[Abstract]
[Full Text]
[PDF]
|
|
|
|
|
|
|
A. Thor
Are Patterns of HER-2/neu Amplification and Expression Among Primary Tumors and Regional Metastases Indicative of Those in Distant Metastases and Predictive of Herceptin Response?
J Natl Cancer Inst,
August 1, 2001;
93(15):
1120 - 1121.
[Full Text]
[PDF]
|
|
|
|
