Abstract P1-02-02: Examination of low Her2 expression in breast cancer
Authors: Fernandez Aileen I , Liu Matthew , Bellizzi Andrew , Brock Jane , Fadare Oluwole , Hanley Krisztina , Harigopal Malini , Jorns Julie M. , Kuba M. Gabriela , Ly Amy , Podoll Mirna , Rabe Kimmie , Sanders Mary Ann , Singh Kamaljeet , Snir Olivia L , Soong Rinda , Wei Shi , Wen Hannah , Wong Serena , Yoon Esther , Pusztai Lajos , Reisenbichler Emily , Rimm David L. 
Published: March 2022

Abstract
Introduction: Antibody-drug conjugates (ADC) are designed to effectively deliver cytotoxic agents directly to malignant cells. Trastuzumab deruxtecan (Tdx), an ADC of trastuzumab, an enzyme-cleavable linker, and a cytotoxic topoisomerase I inhibitor, has been shown to have antitumor activity in patients with breast cancer with low levels of HER2. However, current companion diagnostic tests for HER2-targetting therapies, immunohistochemistry (IHC) and fluorescent in situ hybridization (FISH), were optimized for high (gene amplified) levels of HER2. We hypothesize that the current common assays used in clinic do not efficiently differentiate between patients whose cancers have “0” HER2 expression and “1+” HER2 expression and thus could miss patients who would benefit from treatment with this drug. Methods: Here, we evaluated two years of HER2 surveys from the College of American Pathologists’ (CAP) Proficiency Testing Surveys for HER2 expression in breast cancer from 2019 and 2020. Participating laboratories received two tissue microarrays (TMAs) of 10 breast cancer cores, each Laboratories stained for HER2 using the standard IHC assay used in their labs. The scores were returned to the CAP as part of their quality assessment program. Each survey dataset covers the scores from around 1400 labs of 20 cores each as well as supplemental questions regarding the methodology used. We summarized the relative frequency and distribution of each score given to every core. A second independent analytic dataset was selected from the archives of the Department of Pathology at Yale, from breast biopsies in 2018. The set, enriched in HER2 2+ and 3+ cases, was read by eighteen board-certified pathologists, most with over 5 years’ experience, participating in an interobserver variability study. Hematoxylin and eosin (H&E) and HER2 IHC digitally scanned images of 170 independent cases were provided. Pathologists scored the cases as 0,1, 2, or 3+. Fisher’s exact test was used to compare the 0/1+ concordant cases to 2+/3+ cases. All tests were two-sided at a significant level 0.05. Statistical analysis was performed using Graphpad Prism Version 9.0.1 and the dplyr package in R Version 1.0.143. This study was approved by Yale Human Investigation IRB protocol ID 9505008219. Results: We found that 65% of the 80 cores evaluated in the CAP survey (52/80) had a concordance rate ≥90%. This high concordance was limited to scores of 0 and 3+. The lowest concordance was found between 0 versus 1+. Of the 80 cores, 56 were considered negative (HER2 score of 0 or 1). In 25% of those cores there was &lt; 70% concordance (n=15; 6 in 2019 and 9 in 2020). Analytic concordance was assessed in the independent, Yale cohort where we found that of the 170 cases, 92 were read as 0 by at least one pathologist. Of these 92, 24 were concordant (26%), defined as a ≥90% agreement. In comparison, 45/170 were read as 3+ by at least one pathologist. Again,. using a 90%definition of concordance, 26 of 45 cases (58%) were concordant. Comparison of 0/1+ concordant cases versus 2+/3+ concordant cases showed a significant difference (χ2 = 12.07, p&lt;0.0005). Conclusions: Assessment of laboratory performance of around 1400 CAP labs using common current HER2 assays on CAP survey specimens, there is significant discordance in the evaluation of 0 vs. 1+ cases. In a separate selected breast biopsy cohort examined by 18 breast pathologists, we showed that discordance between scores of 0 vs 1+ is significantly larger than that between 2+ and 3+. Given the efficacy of T-DXd, we believe patients may be mis-assigned for treatment or no treatment if the decision depends on performance of the standard current HER2 assays.
Citation Format: Aileen I Fernandez, Matthew Liu, Andrew Bellizzi, Jane Brock, Oluwole Fadare, Krisztina Hanley, Malini Harigopal, Julie M. Jorns, M. Gabriela Kuba, Amy Ly, Mirna Podoll, Kimmie Rabe, Mary Ann Sanders, Kamaljeet Singh, Olivia L Snir, Rinda Soong, Shi Wei, Hannah Wen, Serena Wong, Esther Yoon, Lajos Pusztai, Emily Reisenbichler, David L. Rimm. Examination of low Her2 expression in breast cancer [abstract]. In: Proceedings of the 2021 San Antonio Breast Cancer Symposium; 2021 Dec 7-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2022;82(4 Suppl):Abstract nr P1-02-02.