Epidermal Growth Factor Receptor (EGFR) High Gene Copy Number and Activating Mutations in Lung Adenocarcinomas Are Not Consistently Accompanied by Positivity for EGFR Protein by Standard Immunohistochemistry

Pintér, Ferenc; Pápay, Judit; Almási, Andrea; Szepes, Zoltán; Szabó, Edit; Kánya, Melinda; Tamási, Anna; Jóri, Balázs; Várkondi, Edit; Moldvay, Judit; Szondy, Klára; Kéri, Gÿorgý; Dominici, Massimo; Conte, Pierfranco; Eckhardt, S.; Kopper, László; Schwab, Richárd; Peták, István

doi:10.2353/jmoldx.2008.070125

Cited by 58 publications

(48 citation statements)

References 31 publications

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“…10,34 However, in the present study, total EGFR was detected in all patients who were proved to harbor mutated EGFR. A novel antibody for total EGFR was used that has not been used in previous studies.…”

Section: Discussioncontrasting

confidence: 68%

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Standardization of Epidermal Growth Factor Receptor (EGFR) Measurement by Quantitative Immunofluorescence and Impact on Antibody-Based Mutation Detection in Non–Small Cell Lung Cancer

Dimou

Agarwal

Anagnostou

et al. 2011

The American Journal of Pathology

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Challenges in measurement of epidermal growth factor receptor (EGFR) protein expression have led to conflicting data on its prognostic value and discontinuation of its use for prediction of response. Herein is described a quantitative standardized assay for EGFR and its use in a series of retrospective cohorts of patients with nonsmall cell lung cancer (NSCLC). The AQUA technology of quantitative immunofluorescence was used in conjunction with Western blot analysis to calculate the absolute concentration of EGFR in two independent NSCLC cohorts (170 from Yale New Haven Hospital and 335 from Sotiria and Patras Hospitals in Greece). EGFR and mutated EGFR were measured using D38B1 antibody and two mutation-specific antibodies. All patients positive or borderline for mutation-specific antibody were genotyped. A threshold for reproducible detection of EGFR was defined as 0.85 ng/ g total protein. EGFR expression demonstrated no prognostic value in either cohort. The mutation rate was 1.79% in the Yale cohort, and 1.52% in the Sotiria/Patras cohort, with no antibody detection-based false-positive cases. No mutations were detected for EGFR concentrations <1.46 ng/ g total protein. In summary, accurate measurement of EGFR still shows no prognostic value in NSCLC. In these two population-based cohorts, the antibody-based EGFR mutation rate was lower than has been frequently reported.

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Section: Discussioncontrasting

confidence: 68%

“…Clones 31G7 and 18C9 were used in previous studies. 10,34 Both detect the extracellular domain of EGFR and require protease-mediated antigen retrieval. In our previous work, these and other EGFR antibodies have been historically difficult to validate.…”

Section: Discussionmentioning

confidence: 99%

Standardization of Epidermal Growth Factor Receptor (EGFR) Measurement by Quantitative Immunofluorescence and Impact on Antibody-Based Mutation Detection in Non–Small Cell Lung Cancer

Dimou

Agarwal

Anagnostou

et al. 2011

The American Journal of Pathology

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“…46 Another study confi rmed that mutational status is the most valuable predictor of responsiveness, independent of copy number or protein expression. 47 Several phase 2 and 3 trials have been conducted to answer this question, but the results have been inconsistent. 48,49 Two meta-analyses confi rmed that EGFR mutational analysis, not FISH, is the assay of choice to predict response to TKI therapy.…”

Section: Methylationmentioning

confidence: 99%

Molecular Biology of Lung Cancer

Nana‐Sinkam

Powell

2013

Chest

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Lung cancer is the leading cause of cancer death in the United States, with 157,000 deaths annually, and in the world, with 1,375,000 deaths annually. 2 The overall 5-year survival for lung cancer remains at 16%, which is signifi cantly lower than the mortality rate for other common cancers, including colon, breast, and prostate cancer. The discrepancy in the mortality rate of lung cancer compared with other cancers has several explanations. First, lung cancers are typically detected at an advanced stage. The anticipated implementation of population-based screening for lung cancer is expected to decrease mortality. Second, compared with the other common cancers that share adenocarcinoma as the histology, lung cancer is heterogenous histologically and biologically. Until recently, lung cancer was treated as a homogenous disease with all nonsmall cell lung cancers (NSCLCs) treated identically, solely on the basis of clinical stage. Based on bench and clinical research, the treatment of lung cancer has been refi ned, with treatments allocated according to histology and specifi c molecular features. For example, targeting mutations such as epidermal growth factor receptor (EGFR) with tyrosine kinase inhibitors (TKIs) has been particularly successful as a treatment modality, demonstrating response rates in patients with EGFR-mutated adenocarcinoma that are significantly higher than those for conventional chemotherapy. However, the development of new targeted therapies is, in part, highly dependent on an improved understanding of the molecular underpinnings of tumor initiation and progression, knowledge of the role of molecular aberrations in disease progression, Based on recent bench and clinical research, the treatment of lung cancer has been refi ned, with treatments allocated according to histology and specifi c molecular features. For example, targeting mutations such as epidermal growth factor receptor (EGFR) with tyrosine kinase inhibitors has been particularly successful as a treatment modality, demonstrating response rates in selected patients with adenocarcinoma tumors harboring EGFR mutations that are signifi cantly higher than those for conventional chemotherapy. However, the development of new targeted therapies is, in part, highly dependent on an improved understanding of the molecular underpinnings of tumor initiation and progression, knowledge of the role of molecular aberrations in disease progression, and the development of highly reproducible platforms for high-throughput biomarker discovery and testing. In this article, we review clinically relevant research directed toward understanding the biology of lung cancer. The clinical purposes of this research are (1) to identify susceptibility variants and fi eld molecular alterations that will promote the early detection of tumors and (2) to identify tumor molecular alterations that serve as therapeutic targets, prognostic biomarkers, or predictors of tumor response. We focus on research developments in the understanding of lung cancer somatic DNA mutat...

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“…IHC for total EGFR is an especially poor substitute as it correlates poorly or not at all with the presence of mutations. 11,12 Another more challenging IHC strategy is to develop antibodies that react only with the mutant form of a given oncoprotein. Interest in this approach is driven by the fact that IHC is a technology available to essentially all pathology departments, can be automated, and can be performed on samples where the number or proportion of tumor cells poses challenges for molecular tests based on bulk DNA extraction from tissue.…”

mentioning

confidence: 99%

Assessment of EGFR Mutation Status in Lung Adenocarcinoma by Immunohistochemistry Using Antibodies Specific to the Two Major Forms of Mutant EGFR

Brevet

Arcila

Ladanyi

2010

The Journal of Molecular Diagnostics

183

162

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EGFR mutations are the best predictors of response to EGFR kinase inhibitors in lung adenocarcinoma. We evaluated two mutation-specific monoclonal antibodies for the detection of EGFR mutations by immunohistochemistry (IHC) , generated respectively against the L858R mutant and the exon 19 mutant with the common 15bp/5AA deletion. These two mutations account for approximately 90% of all EGFR mutations. IHC staining performed on 218 paraffin-embedded lung adenocarcinomas was assessed on a 0 to 3؉ scale , and positivity cutoffs of 1؉ and 2؉ were compared. All cases were studied by standard molecular methods for these two mutations , and selected cases were also studied using higher sensitivity molecular assays. The EGFR L858R mutant antibody showed a sensitivity of 95% and a positive predictive value (PPV) of 99% with a positivity cutoff of 1؉ and a sensitivity of 76% and a PPV of 100% with a positivity cutoff of 2؉. The EGFR exon 19 mutant-specific antibody showed reduced sensitivity for exon 19 deletions other than 15bp. A positivity cutoff of 1؉ resulted in a sensitivity of 85% and a PPV of 99% , whereas a 2؉ cutoff gave a sensitivity of 67% and a PPV of 100%. IHC with EGFR mutant-specific antibodies could be used as a screen to identify most candidates for EGFR inhibitors. (J Mol Diagn

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Epidermal Growth Factor Receptor (EGFR) High Gene Copy Number and Activating Mutations in Lung Adenocarcinomas Are Not Consistently Accompanied by Positivity for EGFR Protein by Standard Immunohistochemistry

Cited by 58 publications

References 31 publications

Standardization of Epidermal Growth Factor Receptor (EGFR) Measurement by Quantitative Immunofluorescence and Impact on Antibody-Based Mutation Detection in Non–Small Cell Lung Cancer

Standardization of Epidermal Growth Factor Receptor (EGFR) Measurement by Quantitative Immunofluorescence and Impact on Antibody-Based Mutation Detection in Non–Small Cell Lung Cancer

Molecular Biology of Lung Cancer

Assessment of EGFR Mutation Status in Lung Adenocarcinoma by Immunohistochemistry Using Antibodies Specific to the Two Major Forms of Mutant EGFR

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