Marius Ilie1,2,3, Véronique Hofman1,2,3, Sylvie Leroy4, Charlotte Cohen5, Simon Heeke3, Florian Cattet6, Coraline Bence1, Salomé Lalvée1, Jérôme Mouroux5, Charles-Hugo Marquette3,4, Paul Hofman1,2,3. 1. Université Côte d'Azur, CHU de Nice, University Hospital Federation OncoAge, Laboratory of Clinical and Experimental Pathology and Liquid Biopsy Laboratory, Nice, France. 2. Université Côte d'Azur, CHU de Nice, University Hospital Federation OncoAge, Hospital-Integrated Biobank (BB-0033-00025), Nice, France. 3. Université Côte d'Azur, CHU de Nice, Institute of Research on Cancer and Ageing of Nice (IRCAN), Inserm U1081, CNRS UMR7284, Team 4, Nice, France. 4. Université Côte d'Azur, CHU de Nice, University Hospital Federation OncoAge, Department of Pulmonary Medicine and Oncology, Nice, France. 5. Université Côte d'Azur, CHU de Nice, University Hospital Federation OncoAge, Department of Thoracic Surgery, Nice, France. 6. Université Côte d'Azur, CHU de Nice, Department of Anesthesiology and Critical Care, Nice, France.
Abstract
BACKGROUND: Circulating tumor cells (CTCs) hold potential for noninvasive diagnosis, prognosis and prediction testing in non-small cell lung cancer (NSCLC) patients. Minimizing degradation or loss of CTCs is pivotal for detection and profiling of the low abundance and fragile CTCs, particularly in clinical trials. We prospectively investigated (NCT02372448) whether a new blood collection device performed better compared to commonly used K3EDTA tubes, when subjected to long-term sample storage. METHODS: Blood samples were drawn into K3EDTA and blood collection tubes (BCT) (Streck), and filtered by the Isolation by SizE of Tumor/Trophoblastic Cells (ISET® system), for CTC detection in two study populations of NSCLC patients; the training set of 14 patients with stage II/IV NSCLC, and the validation set of 36 patients with stage IV NSCLC). MET expression was evaluated by immunocytochemistry (ICC) and anaplastic lymphoma kinase (ALK) gene rearrangement by break-apart fluorescence in situ hybridization (FISH) on ISET-enriched CTCs. RESULTS: Blood processed after 24 h and 48 h in BCT tubes showed stable CTCs counts and integrity, whereas CTCs in K3EDTA tubes showed an altered morphology in all patients. CTCs recovered in BCT or K3EDTA tubes at 24 and 48 h were evaluable by ICC for MET expression and by FISH for ALK rearrangement. CONCLUSIONS: The BCT tubes gave a high yield and preserved the integrity of CTCs after 24 and 48 h of storage at room temperature, which facilitate their molecular characterization in NSCLC patients entering clinical trials.
BACKGROUND: Circulating tumor cells (CTCs) hold potential for noninvasive diagnosis, prognosis and prediction testing in non-small cell lung cancer (NSCLC) patients. Minimizing degradation or loss of CTCs is pivotal for detection and profiling of the low abundance and fragile CTCs, particularly in clinical trials. We prospectively investigated (NCT02372448) whether a new blood collection device performed better compared to commonly used K3EDTA tubes, when subjected to long-term sample storage. METHODS: Blood samples were drawn into K3EDTA and blood collection tubes (BCT) (Streck), and filtered by the Isolation by SizE of Tumor/Trophoblastic Cells (ISET® system), for CTC detection in two study populations of NSCLCpatients; the training set of 14 patients with stage II/IV NSCLC, and the validation set of 36 patients with stage IV NSCLC). MET expression was evaluated by immunocytochemistry (ICC) and anaplastic lymphoma kinase (ALK) gene rearrangement by break-apart fluorescence in situ hybridization (FISH) on ISET-enriched CTCs. RESULTS: Blood processed after 24 h and 48 h in BCT tubes showed stable CTCs counts and integrity, whereas CTCs in K3EDTA tubes showed an altered morphology in all patients. CTCs recovered in BCT or K3EDTA tubes at 24 and 48 h were evaluable by ICC for MET expression and by FISH for ALK rearrangement. CONCLUSIONS: The BCT tubes gave a high yield and preserved the integrity of CTCs after 24 and 48 h of storage at room temperature, which facilitate their molecular characterization in NSCLCpatients entering clinical trials.
Entities:
Keywords:
BCT; Isolation by SizE of Tumor/Trophoblastic Cells (ISET); K3EDTA; circulating tumor cells; non-small cell lung cancer (NSCLC); stability