Related Experiment Video
Updated: May 13, 2025
Validated Immunochemical Assay for Comprehensive Determination of the Human Epidermal Growth Factor Receptor 2 Released from and Bound to Cells
Published on: May 9, 2025
Testing and Interpretation of Human Epidermal Growth Factor Receptor 2 Protein Expression and ERBB2 Gene
Vadim S Koshkin1, Johanna M Schafer2, Emilie Scherrer3
1Division of Hematology/Oncology, Department of Medicine, University of California San Francisco, Helen Diller Family Comprehensive Cancer Center, San Francisco, CA.
Purpose:
The aim of this study was to assess the prevalence of human epidermal growth factor receptor 2 (HER2) protein expression and ERBB2 gene amplification in a large cohort of tumor samples from patients with advanced urothelial carcinoma (UC).
Materials And Methods:
Testing was performed on formalin-fixed, paraffin-embedded tissue from commercially sourced primary advanced or metastatic UC using an anti-HER2/neu (4B5) rabbit monoclonal antibody immunohistochemistry (IHC) assay (detects HER2 protein) and HER2 dual in situ hybridization (DISH) DNA probe cocktail assay (detects ERBB2 gene and the centromere of its residing chromosome 17). The HER2 clinical status was classified as HER2-zero (IHC 0), HER2-low (IHC 1+ or IHC 2+/DISH nonamplified), or HER2-positive (IHC 2+/DISH amplified or IHC 3+).
Results:
Of the 2,024 UC samples, HER2 protein expression was IHC 0 for 962 (47.5%), IHC 1+ for 297 (14.7%), IHC 2+ for 498 (24.6%), and IHC 3+ for 267 (13.2%). The percentage of HER2-expressing tumors (IHC 1+, 2+, and 3+) was similar between primary (52.2%, 1,028/1,968) and metastatic UC samples (60.7%, 34/56; P = .26). The ERBB2 gene was amplified in 235 cases (11.6%), including 2.7%/3.7%/9.6%/56.2% scored as IHC 0/1+/2+/3+, respectively. Overall, HER2 clinical status was HER2-zero for 962 UC tissue samples (47.5%; 95% CI, 45.4 to 49.7), HER2-low for 747 (36.9%; 95% CI, 34.8 to 39.0), and HER2-positive for 315 (15.6%; 95% CI, 14.0 to 17.2).
Conclusion:
We observed that more than 50% of 2,024 advanced UC tumors demonstrated some degree of HER2 protein expression detected using a standardized IHC method, whereas about 12% of specimens had ERBB2 gene amplification, including about 3% of those scored as either IHC 0 or 1+. Continued development of optimized and standardized HER2 testing methods is warranted to identify patients with HER2-expressing UC who may benefit from novel HER2-targeting therapies.
