Swartz Justin E, Smits Hilde J G, Philippens Marielle E P, de Bree Remco, H A M Kaanders Johannes, Willems Stefan M
Department of Otorhinolaryngology - Head and Neck Surgery, University Medical Center Utrecht, Utrecht, the Netherlands; Department of Head and Neck Surgical Oncology, University Medical Center Utrecht, Utrecht, the Netherlands.
Department of Radiotherapy, University Medical Center Utrecht, Utrecht, the Netherlands.
Oral Oncol. 2022 May;128:105862. doi: 10.1016/j.oraloncology.2022.105862. Epub 2022 Apr 18.
Tumor hypoxia results in worse local control and patient survival. We performed a digital, single-cell-based analysis to compare two biomarkers for hypoxia (hypoxia-inducible factor 1-alpha [HIF-1α] and pimonidazole [PIMO]) and their effect on outcome in laryngeal cancer patients treated with accelerated radiotherapy with or without carbogen breathing and nicotinamide (AR versus ARCON).
Immunohistochemical staining was performed for HIF-1α and PIMO in consecutive sections of 44 laryngeal cancer patients randomized between AR and ARCON. HIF-1α expression and PIMO-binding were correlated using digital image analysis in QuPath. High-density areas for each biomarker were automatically annotated and staining overlap was analyzed. Kaplan-Meier survival analyses for local control, regional control and disease-free survival were performed to predict a response benefit of ARCON over AR alone for each biomarker.
106 Tissue fragments of 44 patients were analyzed. A weak, significant positive correlation was observed between HIF-1α and PIMO positivity on fragment level, but not on patient level. A moderate strength correlation (r = 0.705, p < 0.001) was observed between the number of high-density staining areas for both biomarkers. Staining overlap was poor. HIF-1α expression, PIMO-binding or a combination could not predict a response benefit of ARCON over AR.
Digital image analysis to compare positive cell fractions and staining overlap between two hypoxia biomarkers using open-source software is feasible. Our results highlight that there are distinct differences between HIF-1α and PIMO as hypoxia biomarkers and therefore suggest co-existence of different forms of hypoxia within a single tumor.
肿瘤缺氧会导致局部控制不佳和患者生存率降低。我们进行了一项基于数字单细胞的分析，以比较两种缺氧生物标志物（缺氧诱导因子1α [HIF-1α] 和匹莫硝唑 [PIMO]）及其对接受加速放疗联合或不联合卡波金呼吸及烟酰胺治疗的喉癌患者（AR与ARCON）预后的影响。
对44例在AR和ARCON之间随机分组的喉癌患者的连续切片进行HIF-1α和PIMO的免疫组织化学染色。在QuPath中使用数字图像分析将HIF-1α表达和PIMO结合情况进行关联。自动标注每种生物标志物的高密度区域并分析染色重叠情况。进行局部控制、区域控制和无病生存的Kaplan-Meier生存分析，以预测ARCON相对于单独AR对每种生物标志物的反应益处。
分析了44例患者的106个组织切片。在切片水平上观察到HIF-1α和PIMO阳性之间存在微弱的显著正相关，但在患者水平上未观察到。两种生物标志物的高密度染色区域数量之间观察到中等强度的相关性（r = 0.705，p < 0.001）。染色重叠较差。HIF-1α表达、PIMO结合或两者结合均无法预测ARCON相对于AR的反应益处。
使用开源软件通过数字图像分析比较两种缺氧生物标志物的阳性细胞分数和染色重叠是可行的。我们的结果表明，HIF-1α和PIMO作为缺氧生物标志物存在明显差异，因此提示单个肿瘤内存在不同形式的缺氧共存情况。
