Single-Cell Quantitative HER2 Measurement Identifies Heterogeneity and Distinct Subgroups within Traditionally Defined HER2-Positive Patients
Single-Cell Quantitative HER2 Measurement Identifies Heterogeneity and Distinct Subgroups within Traditionally Defined HER2-Positive Patients
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DOI：
10.1016/j.ajpath.2013.07.015
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发表时间：
2013-11-01
影响因子：
6
通讯作者：
Hendriks, Bart S.
中科院分区：
文献类型：
--
作者：
Onsum, Matthew D.;Geretti, Elena;Hendriks, Bart S.
人表皮生长因子受体 2 (HER2) 是乳腺癌和胃癌预后和患者治疗决策的重要生物标志物。根据 IHC 或荧光原位杂交检测的定义，HER2 阳性仍然是患者对 HER2 靶向治疗反应的不精确预测指标。纠正 HER2 评估和患者分层的挑战包括瘤内异质性、缺乏定量和/或客观测定，以及在蛋白质水平和基因水平测量 HER2 扩增之间的差异。我们开发了一种新型免疫荧光方法，用于在 FFPE 患者样本的单细胞水平上定量 HER2 蛋白表达。我们的检测使用自动图像分析来识别和分类肿瘤细胞与非肿瘤细胞，并对每个肿瘤细胞的 HER2 染色进行定量。使用与组织样本平行染色的标准细胞沉淀阵列将 HER2 染色水平转换为 HER2 蛋白表达。这种方法可以在单细胞水平上评估组织切片内的 HER2 表达和异质性。通过使用这种测定，我们在乳腺癌和胃癌中 HER2 阳性的传统定义中确定了 HER2 异质性的不同亚组。肿瘤内 HER2 异质性的定量评估可能为改善对可能对 HER2 靶向治疗产生反应的患者的识别提供机会。通过测量多种肿瘤类型以及正常和患病心脏组织的 HER2 表达谱，证明了该测定法的广泛适用性。
Human epidermal growth factor receptor 2 (HER2) is an important biomarker for breast and gastric cancer prognosis and patient treatment decisions. HER2 positivity, as defined by IHC or fluorescent in situ hybridization testing, remains an imprecise predictor of patient response to HER2-targeted therapies. Challenges to correct HER2 assessment and patient stratification include intratumoral heterogeneity, lack of quantitative and/or objective assays, and differences between measuring HER2 amplification at the protein versus gene level. We developed a novel immunofluorescence method for quantitation of HER2 protein expression at the single-cell level on FFPE patient samples. Our assay uses automated image analysis to identify and classify tumor versus non-tumor cells, as well as quantitate the HER2 staining for each tumor cell. The HER2 staining level is converted to HER2 protein expression using a standard cell pellet array stained in parallel with the tissue sample. This approach allows assessment of HER2 expression and heterogeneity within a tissue section at the single-cell level. By using this assay, we identified distinct subgroups of HER2 heterogeneity within traditional definitions of HER2 positivity in both breast and gastric cancers. Quantitative assessment of intratumoral HER2 heterogeneity may offer an opportunity to improve the identification of patients likely to respond to HER2-targeted therapies. The broad applicability of the assay was demonstrated by measuring HER2 expression profiles on multiple tumor types, and on normal and diseased heart tissues.