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Inter-Assay Variability of TROP2 Immunohistochemistry in Triple-Negative Breast Cancer
Giulia Cursano1,2, Alberto Concardi1,2, Mariia Ivanova1
1Division of Pathology, IEO, European Institute of Oncology IRCCS, Via G. Ripamonti 435, 20141, Milan, Italy.
Molecular Diagnosis & Therapy
|September 6, 2025
Summary
This study found significant variability in TROP2 expression analysis across different immunohistochemistry assays for triple-negative breast cancer. Standardizing these assays is crucial for reliable TROP2 biomarker assessment.
Area of Science:
- Oncology
- Immunohistochemistry
- Biomarker Analysis
Background:
- Sacituzumab govitecan targets TROP2 in metastatic triple-negative breast cancer (TNBC).
- TROP2 expression, assessed by immunohistochemistry (IHC), correlates with improved outcomes but lacks standardized assays.
- This variability impacts the reliable clinical utility of TROP2 as a biomarker.
Purpose of the Study:
- To evaluate the consistency of TROP2 expression analysis across three different IHC assays.
- To assess the concordance of TROP2 quantification using the H-score method.
- To identify the need for standardized IHC protocols in TNBC TROP2 biomarker assessment.
Main Methods:
- Analyzed 26 TNBC samples using three distinct IHC assays (ENZO-ABS380-0100, Abcam SP295, Santa Cruz B9-sc-376746) on a Dako Omnis platform.
- Quantified TROP2 membrane expression using the H-score, categorizing results as low, intermediate, or high.
- Evaluated assay agreement using Cohen's κ and Gwet's AC2 statistics.
Main Results:
- Significant variability in TROP2 expression classification was observed across the three assays.
- Assays B and C showed substantial agreement (80.8% concordance, κ = 0.81), while overall concordance between all three assays was fair to moderate (AC2 = 0.35).
- Assay A demonstrated a broader range of TROP2 expression compared to assays B and C.
Conclusions:
- There are significant inter-assay variations in TROP2 detection, impacting its reliability as a biomarker in TNBC.
- TROP2 expression is heterogeneous and dynamic, necessitating methodological improvements in IHC testing.
- Standardized IHC protocols and quantitative scoring systems, potentially integrated with computational pathology, are needed to enhance TROP2's clinical utility.
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