Single tube liquid biopsy for advanced non‐small cell lung cancer

Abstract

The need for a liquid biopsy in non‐small cell lung cancer (NSCLC) patients is rapidly increasing. We studied the relation between overall survival (OS) and the presence of four cancer biomarkers from a single blood draw in advanced NSCLC patients: EpCAMhigh circulating tumor cells (CTC), EpCAMlow CTC, tumor‐derived extracellular vesicles (tdEV) and cell‐free circulating tumor DNA (ctDNA). EpCAMhigh CTC were detected with CellSearch, tdEV in the CellSearch images and EpCAMlow CTC with filtration after CellSearch. ctDNA was isolated from plasma and mutations present in the primary tumor were tracked with deep sequencing methods. In 97 patients, 21% had ≥2 EpCAMhigh CTC, 15% had ≥2 EpCAMlow CTC, 27% had ≥18 tdEV and 19% had ctDNA with ≥10% mutant allele frequency. Either one of these four biomarkers could be detected in 45% of the patients and all biomarkers were present in 2%. In 11 out of 16 patients (69%) mutations were detected in the ctDNA. Two or more unfavorable biomarkers were associated with poor OS. The presence of EpCAMhigh CTC and elevated levels of tdEV and ctDNA was associated with a poor OS; however, the presence of EpCAMlow CTC was not. This single tube approach enables simultaneous analysis of multiple biomarkers to explore their potential as a liquid biopsy.

Detection of Circulating Tumor Cells in the Diagnostic Leukapheresis Product of Non-Small-Cell Lung Cancer Patients Comparing CellSearch® and ISET

“…EpCAM+ CTCs were detected in similar proportions of patients and in similar concentrations by CellSearch and ISET. As EpCAM+ CTCs are possibly more strongly associated with clinical outcome, both CellSearch and ISET function well for CTCs that have been proven to be both predictive and prognostic [5]. However, due to the larger volume processed by ISET, this procedure can isolate a larger number of CTCs for further functional or genomic analysis.…”

“…When detected in sufficient numbers, they can be used for molecular analysis. Unfortunately, CTCs are only detected in 30% of NSCLC patients and usually in low numbers, hampering their clinical application [5,6]. It is likely that the majority of metastatic patients have CTCs in circulation, but that the volume of blood screened for CTCs (7.5 mL) is insufficient for a reliable detection [7].…”

Tumour-derived extracellular vesicles in blood of metastatic cancer patients associate with overall survival

“…One hundred and ninety CRPC (IMMC38 clinical trial, NCT00133900), 30 450 mCRC (CAIRO II clinical trial, NCT00208546), 31,32 179 MBC (IMMC01 clinical trial), 33 and 137 NSCLC patient samples, [34][35][36] along with 93 healthy control samples (IMMC06 clinical trial, NCT00133913), 37 were included. The included samples corresponded to patients before the initiation of a new treatment.…”

“…All individuals provided written informed consent prior to participation in the trial protocols approved by institutional review boards at the participating centres of the studies. [31][32][33][34][35][36][37][38] Isolation and identification of CTCs and tdEVs Digitally stored CellSearch® (Menarini Silicon Biosystems, Huntingdon Valley, PA, USA) image files from the abovementioned CTC studies were re-analysed. Briefly, CTCs and tdEVs were immunomagnetically isolated from 7.5 mL of peripheral blood collected in Cell Save tubes using the CellSearch system.…”

“…Ethics approval and consent to participate All individuals provided written informed consent prior to participation in the trial protocols approved by institutional review boards at the participating centres of the studies. [31][32][33][34][35][36][37][38] The studies were performed in accordance with the Declaration of Helsinki.…”