Introduction: Phosphatase and tensin homolog (PTEN) loss is frequently observed in NSCLC and associated with both phosphoinositide 3‐kinase activation and tumoral immunosuppression. PTEN immunohistochemistry is a valuable readout, but lacks standardized staining protocol and cutoff value. Methods: After an external quality assessment using SP218, 138G6 and 6H2.1 anti‐PTEN antibodies, scored on webbook and tissue microarray, the European Thoracic Oncology Platform cohort samples (n = 2245 NSCLC patients, 8980 tissue microarray cores) were stained with SP218. All cores were H‐scored by pathologists and by computerized pixel‐based intensity measurements calibrated by pathologists. Results: All three antibodies differentiated six PTEN+ versus six PTEN‐ cases on external quality assessment. For 138G6 and SP218, high sensitivity and specificity was found for all H‐score threshold values including prospectively defined 0, calculated 8 (pathologists), and calculated 5 (computer). High concordance among pathologists in setting computer‐based intensities and between pathologists and computer in H‐scoring was observed. Because of over‐integration of the human eye, pixel‐based computer H‐scores were overall 54% lower. For all cutoff values, PTEN‐ was associated with smoking history, squamous cell histology, and higher tumor stage (p < 0.001). In adenocarcinomas, PTEN‐ was associated with poor survival. Conclusion: Calibration of immunoreactivity intensities by pathologists following computerized H‐score measurements has the potential to improve reproducibility and homogeneity of biomarker detection regarding epitope validation in multicenter studies.

中文翻译：

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基于计算机的强度测量可帮助病理学家对 PTEN 免疫组织化学进行评分 - ETOP Lungscape 队列中 NSCLC 患者的临床关联

简介：在 NSCLC 中经常观察到磷酸酶和张力蛋白同源物 (PTEN) 的丢失，并且与磷酸肌醇 3-激酶激活和肿瘤免疫抑制有关。PTEN 免疫组织化学是一种有价值的读数，但缺乏标准化的染色方案和临界值。方法：在使用 SP218、138G6 和 6H2.1 抗 PTEN 抗体进行外部质量评估后，在网络书和组织微阵列上评分后，欧洲胸科肿瘤平台队列样本（n = 2245 名 NSCLC 患者，8980 个组织微阵列核心）用 SP218 染色. 所有核心均由病理学家和病理学家校准的基于计算机像素的强度测量进行 H 评分。结果：在外部质量评估中，所有三种抗体都区分了 6 个 PTEN+ 和 6 个 PTEN- 病例。对于 138G6 和 SP218，所有 H 分数阈值都具有高灵敏度和特异性，包括前瞻性定义的 0、计算值 8（病理学家）和计算值 5（计算机）。观察到病理学家在设置基于计算机的强度以及病理学家和计算机之间在 H 评分方面的高度一致性。由于人眼的过度整合，基于像素的计算机 H 分数总体上降低了 54%。对于所有临界值，PTEN-与吸烟史、鳞状细胞组织学和较高的肿瘤分期相关（p < 0.001）。在腺癌中，PTEN-与较差的存活率相关。结论：病理学家在计算机化 H 分数测量后校准免疫反应性强度有可能提高多中心研究中表位验证生物标志物检测的可重复性和同质性。