Quantitative comparison of immunohistochemical HER2-low detection in an interlaboratory study

IF 3.9 2区医学 Q2 CELL BIOLOGY

Histopathology Pub Date : 2024-07-29 DOI:10.1111/his.15273

Maaike Anna Hempenius, Maran A Eenkhoorn, Henrik Høeg, David J Dabbs, Bert van der Vegt, Seshi R Sompuram, Nils A ‘t Hart

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引用次数: 0

Abstract

Aims

Recently, human epidermal growth factor 2 (HER2)-low (i.e. HER2 score 1+ or 2+ without amplification) breast cancer patients became eligible for trastuzumab–deruxtecan treatment. To improve assay standardisation and detection of HER2-low in a quantitative manner, we conducted an external quality assessment-like study in the Netherlands. Dynamic range cell lines and immunohistochemistry (IHC) calibrators were used to quantify HER2 expression and to assess interlaboratory variability.

Methods and results

Three blank slides with a dynamic range cell line and an IHC calibrator were stained with routine HER2 assays by 35 laboratories. Four different antibody clones were used: 19 (54.3%) 4B5, six (17.1%) A0485, five (14.3%) DG44 (HercepTest) and five (14.3%) SP3. Laboratories used two different detection kits for 4B5 assays: 14 (73.7%) ultraView and five (26.3%) OptiView. Variability of HER2 expression in cell lines, measured with artificial intelligence software, was median (min–max) = negative core 0.5% (0.0–57.0), 1+ core 4.3% (1.6–71.3), 2+ core 42.8% (30.4–92.6) and 3+ core 96.2% (91.8–98.8). The calibrators DG44 and 4B5 OptiView had the highest analytical sensitivity, closely followed by 4B5 ultraView. SP3 was the least sensitive. Calibrators of A0485 assays were not analysable due to background staining.

Conclusions

As assays were validated for detecting HER2-amplified tumours, not all assays and antibodies proved suitable for HER2-low detection. Some tests showed distinct expression in the negative cell line. Dynamic range cell line controls and quantitative analysis using calibrators demonstrated more interlaboratory variability than commonly appreciated. Revalidation of HER2 tests by laboratories is needed to ensure clinical applicable HER2-low assays.

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实验室间研究中免疫组化 HER2 低检测的定量比较。

目的：最近，低人类表皮生长因子 2（HER2）（即 HER2 评分 1+ 或 2+，无扩增）乳腺癌患者开始有资格接受曲妥珠单抗-德鲁司康治疗。为了提高检测的标准化程度和定量检测 HER2-low，我们在荷兰开展了一项类似外部质量评估的研究。我们使用动态范围细胞系和免疫组化（IHC）校准物来定量检测HER2的表达，并评估实验室间的差异性：35 家实验室用常规 HER2 检测法对带有动态范围细胞系和 IHC 校准物的三张空白切片进行染色。使用了四种不同的抗体克隆：19个（54.3%）4B5、6个（17.1%）A0485、5个（14.3%）DG44（HercepTest）和5个（14.3%）SP3。实验室使用两种不同的检测试剂盒进行 4B5 检测：14 种（73.7%）ultraView 和 5 种（26.3%）OptiView。用人工智能软件测量细胞系中 HER2 表达的变异性，中位数（最小值-最大值）= 阴性核心 0.5% (0.0-57.0)，1+ 核心 4.3% (1.6-71.3)，2+ 核心 42.8% (30.4-92.6) 和 3+ 核心 96.2% (91.8-98.8)。校准器 DG44 和 4B5 OptiView 的分析灵敏度最高，紧随其后的是 4B5 ultraView。SP3 的灵敏度最低。A0485 检测方法的校准物因背景染色而无法分析：结论：由于检测HER2-扩增肿瘤的检测方法已经过验证，因此并非所有的检测方法和抗体都适用于HER2-低表达肿瘤的检测。一些检测方法在阴性细胞系中有明显的表达。动态范围细胞系对照和使用校准物进行的定量分析显示，实验室之间的变异性比人们通常认为的要大。实验室需要对 HER2 检测进行重新验证，以确保 HER2 低检测适用于临床。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Histopathology 医学-病理学

CiteScore

10.20

自引率

4.70%

发文量

239

审稿时长

1 months

期刊介绍： Histopathology is an international journal intended to be of practical value to surgical and diagnostic histopathologists, and to investigators of human disease who employ histopathological methods. Our primary purpose is to publish advances in pathology, in particular those applicable to clinical practice and contributing to the better understanding of human disease.