■口腔鳞状细胞癌(OSCC)约占口腔恶性肿瘤的90%,5年死亡率接近50%。所有口腔癌的一致部分(70%)在晚期被诊断,因为可用的筛查技术是无效的。因此,迫切需要改进它们。诊断金标准是具有组织学和免疫组织化学评估的组织活检。这种方法存在一些局限性。活检是侵入性的,组织病理学评估是半定量的,并且不能可靠地确定目标的绝对丰度。此外,组织是高度加工的,可能导致自然状态的信息丢失。在用细胞刷收集的组织/细胞碎片上寻找经典和新的临床生物标志物是一种非常有希望的早期检测和诊断OSCC的技术。由于其无创采样和易于收集的方法。
■在这里,我们通过应用创新的高灵敏度ELISA技术,分析了从15例已经诊断出OSCC的患者口腔中收集的细胞刷活检样本,为了验证这种方法是否可以为检测提供有用的信息,诊断,OSCC的预后。为此,我们选择了六种生物标志物,已经在临床实践中用于诊断OSCC(EGFR,Ki67,p53)或根据最近的科学和临床数据进行选择,这些数据表明它们在正在发生转化的细胞中的存在或过表达,以及它们在免疫刺激点阻断疗法中可能作为分子靶标的作用(PD-L1,HLA-E,B7-H6).
■所选择的肿瘤生物标志物在肿瘤核心中高表达,而从同一患者收集的健康组织几乎为阴性。这些差异具有高度统计学意义,并且与使用黄金标准测试获得的差异一致,清楚地表明所提出的方法,即通过定制ELISA技术分析生物标志物,非常可靠。
■这些初步数据表明,这种非侵入性快速表型鉴定技术可用作口腔病变表型鉴定的筛查工具,并通过对病变特征的精确指示来支持临床实践,还着眼于新的抗肿瘤治疗方法的应用,比如免疫疗法,针对OSCC患者。
Oral squamous cell carcinoma (OSCC) accounts for approximately 90% of oral malignancies and has a 5-year mortality rate close to 50%. A consistent part (70%) of all oral cancers is diagnosed at an advanced stage since available screening techniques are ineffective. Therefore, it would be urgent to improve them. The diagnostic gold standard is tissue biopsy with histological and immunohistochemical assessment. This method presents some limitations. Biopsy is invasive and the histopathological evaluation is semi-quantitative, and the absolute abundance of the target cannot be reliably determined. In addition, tissue is highly processed and may lead to loss of information of the natural state. The search for classical and new clinical biomarkers on fragments of tissue/cells collected with a cytobrush is a highly hopeful technique for early detection and diagnosis of OSCC, because of its non-invasive sampling and easy collection method.
Here we analyzed cytobrush biopsies samples collected from the oral cavity of 15 patients with already diagnosed OSCC by applying an innovative high-sensitivity ELISA technique, in order to verify if this approach may provide useful information for detection, diagnosis, and prognosis of OSCC. To this end, we selected six biomarkers, already used in clinical practice for the diagnosis of OSCC (EGFR, Ki67, p53) or selected based on recent scientific and clinical data which indicate their presence or over-expression in cells undergoing transformation and their role as possible molecular targets in immunecheckpoints blockade therapies (PD-L1, HLA-E, B7-H6).
The selected tumor biomarkers were highly expressed in the tumor core, while were virtually negative in healthy tissue collected from the same patients. These differences were highly statistically significant and consistent with those obtained using the gold standard test clearly indicating that the proposed approach, i.e. analysis of biomarkers by a custom ELISA technique, is strongly reliable.
These preliminary data suggest that this non-invasive rapid phenotyping technique could be useful as a screening tool for phenotyping oral lesions and support clinical practice by precise indications on the characteristics of the lesion, also with a view to the application of new anti-tumor treatments, such as immunotherapy, aimed at OSCC patients.