一种用于检测循环肿瘤细胞（CTC）中泛细胞角蛋白和EML4-ALK融合蛋白表达的全自动检测方法可预测非小细胞肺癌（NSCLC）患者的预后。

A fully automated assay to detect the expression of pan-cytokeratins and of EML4-ALK fusion protein in circulating tumour cells (CTCs) predicts outcome of non-small cell lung cancer (NSCLC) patients.

作者信息

Rossi Elisabetta, Aieta Michele, Tartarone Alfredo, Pezzuto Aldo, Facchinetti Antonella, Santini Daniele, Ulivi Paola, Ludovini Vienna, Possidente Luciana, Fiduccia Pasquale, Minicuci Nadia, Zamarchi Rita

机构信息

Department of Surgery, Oncology and Gastroenterology, University of Padova, Padova, Italy.

Veneto Institute of Oncology IOV - IRCCS, Padua, Italy.

出版信息

Transl Lung Cancer Res. 2021 Jan;10(1):80-92. doi: 10.21037/tlcr-20-855.

BACKGROUND

In advanced non-small cell lung cancer (NSCLC) a recent meta-analysis confirms circulating tumour cells (CTCs) as an independent prognostic indicator of progression-free survival (PFS) and overall survival (OS). However, further investigations are necessary to predict and dynamically monitor the therapy in NSCLC patients using CTCs. To this aim, we combined the classical standard assay (SA) with an expanded cytokeratins profile (EA) and quantified the expression of EML4-ALK fusion protein in CTCs.

METHODS

The CellSearch (CS) platform-first marked diagnostic use (IVD) from Food and Drug Administration (FDA), and "gold standard" for quantifying CTCs - detects EpCAM and cytokeratins (CKs) 8, 18, and 19. Since NSCLC shows different CKs profile, we customized the SA, to recognize CK 4, 5, 6, 7, 8, 10, 13, 14, 18, and 19 (EA). Using both assays we designed a prospective, multi-center study, primarily aimed to enumerate CTCs in advanced NSCLC. Secondarily, we developed an integration of the EA to quantify the expression of EML4-ALK fusion protein in CTCs, and correlated them with PFS and OS.

RESULTS

EA identified a significantly much more number of CTC-positive patients (115 out of 180) than SA (103 out of 192; Chi-square =4.0179, with 1 degrees of freedom, P=0.04502). Similar to SA, EA levels were still associated with patient' outcomes. Furthermore, the expression of EML4-ALK on CTCs allowed stratifying NSCLC patients according to a statistically significant difference in PFS.

CONCLUSIONS

We proposed here two novel automated tests, to characterize the expression of specific molecules on CTCs. We demonstrated that these integrated assays are robust and actionable in prospective clinical studies, and in the future could allow clinicians to improve both choice and length of treatment in individual NSCLC patient.

背景

在晚期非小细胞肺癌（NSCLC）中，最近的一项荟萃分析证实循环肿瘤细胞（CTC）是无进展生存期（PFS）和总生存期（OS）的独立预后指标。然而，有必要进行进一步研究，以利用CTC预测和动态监测NSCLC患者的治疗。为此，我们将经典标准检测法（SA）与扩展的细胞角蛋白谱（EA）相结合，并对CTC中EML4-ALK融合蛋白的表达进行定量分析。

方法

CellSearch（CS）平台是美国食品药品监督管理局（FDA）首个获批用于诊断的体外诊断产品（IVD），也是定量CTC的“金标准”，可检测上皮细胞黏附分子（EpCAM）以及细胞角蛋白（CK）8、18和19。由于NSCLC表现出不同的CK谱，我们定制了SA以识别CK 4、5、6、7、8、10、13、14、18和19（EA）。我们使用这两种检测方法设计了一项前瞻性多中心研究，主要目的是对晚期NSCLC患者的CTC进行计数。其次，我们开发了一种整合EA的方法，以定量CTC中EML4-ALK融合蛋白的表达，并将其与PFS和OS相关联。