Automated chromogenic multiplexed immunohistochemistry assay for diagnosis and predictive biomarker testing in non-small cell lung cancer
Automated chromogenic multiplexed immunohistochemistry assay for diagnosis and predictive biomarker testing in non-small cell lung cancer
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DOI：
10.1016/j.lungcan.2018.07.037
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发表时间：
2018-10-01
期刊：
影响因子：
5.3
通讯作者：
Hofman, Paul
中科院分区：
文献类型：
--
作者：
Ilie, Marius;Beaulande, Melanie;Hofman, Paul
目的：目前在病理学实验室中管理非小细胞肺癌（NSCLC）的挑战是在越来越小的活检上结合联合收割机免疫组织化学（IHC）和分子方法，并且需要保留相当数量的肿瘤材料用于越来越大的面板的分子分析。最新的肺癌分类，特别是在低分化肿瘤的情况下，需要进行IHC检查，以进行准确的诊断，并保留尽可能多的组织用于分子检测。因此，建议尽可能减少使用术语NSCLC，并根据其特定的组织学亚型对肿瘤进行分类。这意味着限制组织载玻片的数量，尽管存在特异性和敏感的生物标志物（ALK，ROS 1，BRAF V600 E，PD-L1）和区分肺腺癌（TTF-1阳性）和鳞状细胞癌（p40阳性）的义务。材料和方法：包括来自18例NSCLC患者的样本，这些患者先前以组织学和基因组/免疫学特征为特征。开发了两种多重免疫组化检测方法，用于诊断和免疫分型，包括TTF 1，p40，PD-L1和泛角蛋白抗体，以及用于分子谱分析面板，包括ALK，ROS 1和BRAF V600 E抗体。结果：我们开发了两种敏感的多重免疫组化检测方法，以全面表征主要NSCLC组织型和FDA批准的预测生物标志物，没有抗原性损失，空间干扰或交叉反应性增加。该检测依赖于标准的抗原检索和自动染色协议，限制了验证strategies.Conclusion的需要：我们的多重IHC方法提供了一个独特的样本保留工具，以表征有限的组织样本在肺肿瘤，并使其成为一种替代方法，在临床环境中的晚期NSCLC的治疗决策，提供了在更大的人口进行验证。
Objectives: The current challenge in the management of non-small cell lung cancer (NSCLC) in pathology laboratories is to combine immunohistochemistry (IHC) and molecular approaches on increasingly smaller biopsies and the need to reserve a fair amount of tumor material for molecular analyses with increasingly larger panels. The latest lung cancer classification, especially in the setting of poorly differentiated tumors, requires an IHC workup to allow for accurate diagnosis and also to preserve as much tissue as possible for molecular testing. Thus, it is recommended to reduce use of the term NSCLC not otherwise specified as much as possible and classify tumors according to their specific histologic subtype. This implies limiting the number of tissue slides despite the existence of specific and sensitive biomarkers (ALK, ROS1, BRAF V600E, PD-L1) and the obligation to distinguish lung adenocarcinoma (TTF-1 positive) from squamous cell carcinoma (p40 positive).Materials and Methods: Samples from 18 patients with NSCLC, previously characterized for histologic and genomic/immune features, were included. Two multiplexed IHC assays were developed, for diagnosis and immunophenotyping including TTF1, p40, PD-L1, and pan-Keratin antibodies, and for molecular profiling panel including ALK, ROS1 and BRAF V600E antibodies.Results: We developed two sensitive multiplexed IHC assays to comprehensively characterize major NSCLC histotypes and FDA-cleared predictive biomarkers, without antigenicity loss, steric interference or increased cross-reactivity. The assays rely on standard antigen retrieval and automated staining protocols, limiting the need for validation strategies.Conclusion: Our multiplexed IHC approach provides a unique sample-sparing tool to characterize limited tissue samples in lung oncology and making it an alternative method in the clinical setting for therapeutic decision making of advanced NSCLC, provided that validation in a larger population is performed.