Fidelity of Next-Generation Sequencing to Predict Human Epidermal Growth Factor 2 Overexpression Across Solid Tumors

PURPOSE Human epidermal growth factor 2 (HER2/ERBB2) has recently become a pan-tumor–agnostic target after the approval of trastuzumab deruxtecan for solid tumors with HER2 overexpression (3+). HER2 positivity is currently assessed by immunohistochemistry (IHC) and/or HER2 amplification through in situ hybridization (ISH), owing to the fact that HER2 overexpression is secondary to HER2 gene amplification in many cases. Outside of breast/gastroesophageal cancer, the optimal IHC scoring method to assess overexpression across solid tumors remains undefined. Next-generation sequencing (NGS) is frequently used in practice and could be leveraged to predict HER2 positivity. However, less is known regarding the correlation between these methodologies. We sought to evaluate the ability of NGS to predict HER2 overexpression. METHODS We compared the concordance of HER2 IHC and/or ISH results with HER2 amplification status detected by NGS in 1,009 patients with solid tumors who were referred to the Indiana University Precision Genomics program between September 2021 and October 2024. RESULTS HER2 3+ by IHC was identified in 4.3% (43 of 1,009) of cases. When including HER2 2+/ISH-positive (ISH+), the overall incidence of HER2 positivity was 5.1% (51 of 1,009). HER2 amplification was not detected by NGS in 20.9% (9 of 43) of HER2 3+ and 75.0% (6 of 8) of HER2 2+/ISH+ cases. The rate of discordance varies across the type of NGS testing platform and sample status, with the highest incidence seen in liquid NGS (80.3%) and the lowest incidence observed when the same sample was used for testing (26.7%). CONCLUSION Relying solely on NGS-based HER2 amplification could result in missing cases of HER2 positivity and thus deprive patients of anti-HER2 therapy options. NGS, IHC, and/or ISH should be used as complementary tools for optimal detection of HER2 positivity.