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Use of circulating tumor cells in prospective clinical trials for NSCLC patients – standardization of the pre-analytical conditions

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Published/Copyright: February 3, 2018
Clinical Chemistry and Laboratory Medicine (CCLM)
From the journal Clinical Chemistry and Laboratory Medicine (CCLM) Volume 56 Issue 6

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.

Keywords: BCT; circulating tumor cells; Isolation by SizE of Tumor/Trophoblastic Cells (ISET); K3EDTA; non-small cell lung cancer (NSCLC); stability
Corresponding author: Prof. Paul Hofman, Université Côte d’Azur, CHU de Nice, University Hospital Federation OncoAge, Laboratory of Clinical and Experimental Pathology and Liquid Biopsy Laboratory, Pasteur Hospital, BP69, 06001 Nice Cedex, France, Phone: +33 4 92 03 88 55, Fax: +33 4 92 03 88 50
aMembers of the AIR project Study Group: Dominique Israel-Biet MD (Hôpital Européen Georges Pompidou), Christophe Pison MD (Hôpital Universitaire de Grenoble), Pascal Chanez MD (Hôpital Universitaire de Marseille), Francois Chabot MD (Hôpital Universitaire de Nancy), Gaetan Deslee MD (Hôpital Universitaire de Reims), Hervé Mal MD (Hôpital Bichat), Romain Kessler MD (Hôpital Universitaire de Strasbourg), Jean-Michel Vergnon MD, Isabelle Pelissier MD (Hôpital Universitaire de St Etienne), Antoine Cuvelier MD (Hôpital Universitaire de Rouen), Arnaud Bourdin MD (Hôpital Universitaire de Montpellier), Vincent Jounieaux MD (Hôpital Universitaire d’Amiens), Nicolas Roche MD (Hôpital Cochin), Stephane Jouneau MD (Hôpital Universitaire de Rennes), Philippe Bonniaud MD (Hôpital Universitaire de Dijon), Arnaud Scherpereel MD (Hôpital Universitaire de Lille), Jean Francois Mornex MD (Hôpital Universitaire de Lyon), Francois Steenhouwer MD (Hôpital de Roubaix), Sylvie Leroy MD, Charles Hugo Marquette MD, Jonathan Benzaquen, Andrea Mazzette MD, Bernard Padovani MD, Paul Hofman MD, Marius Ilie MD, Veronique Hofman MD, Johanna Pradelli MD, Maureen Fontaine, Jennifer Griffonnet, Ariane Guillemart, Catherine Butori MD, Eric Selva, (Hôpital Universitaire de Nice), Sylvain Marchand-Adam MD, Laurent Plantier, Gaelle Fajolle, Melanie Rayez (Hôpital Universitaire de Tours), Jacques Cadranel MD, Vincent Falle MD, Nouha Chaabane MD, Anne Marie Ruppert MD (Hôpital Tenon), Julien Mazières MD, Damien Rouviere MD, Emilie Bousquet MD (Hôpital Universitaire de Toulouse).

  1. Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.

  2. Research funding: STALKLUNG01 (Institut National du Cancer, PHRC National Cancer 2014-A00417-40, Funder Id: 10.13039/501100006364), Cancéropôle PACA, Ligue départementale 06, Conseil départemental 06 Appel à projet santé.

  3. Employment or leadership: None declared.

  4. Honorarium: None declared.

  5. Competing interests: The funding organization(s) played no role in the study design; in the collection, analysis, and interpretation of data; in the writing of the report; or in the decision to submit the report for publication.

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Supplemental Material:

The online version of this article offers supplementary material (https://doi.org/10.1515/cclm-2017-0764).

Received: 2017-8-26
Accepted: 2017-12-26
Published Online: 2018-2-3
Published in Print: 2018-5-24

©2018 Walter de Gruyter GmbH, Berlin/Boston

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https://doi.org/10.1515/cclm-2017-0764
Keywords for this article
BCT; circulating tumor cells; Isolation by SizE of Tumor/Trophoblastic Cells (ISET); K3EDTA; non-small cell lung cancer (NSCLC); stability

Articles in the same Issue

  1. Frontmatter
  2. Editorial
  3. Developments in laboratory testing for autoimmune diseases
  4. Reviews
  5. Cerebrospinal fluid microRNAs as diagnostic biomarkers in brain tumors
  6. HCV core antigen comes of age: a new opportunity for the diagnosis of hepatitis C virus infection
  7. Mini Reviews
  8. Insulin autoimmune syndrome (Hirata’s disease) in an Italian patient: a case report and review of the literature
  9. The natural history of autoimmune Addison’s disease with a non-classical presentation: a case report and review of literature
  10. Opinion Paper
  11. Quality and future of clinical laboratories: the Vico’s whole cyclical theory of the recurring cycles
  12. General Clinical Chemistry and Laboratory Medicine
  13. Screening for connective tissue disease-associated antibodies by automated immunoassay
  14. Analysis of anti-ganglioside antibodies by a line immunoassay in patients with chronic-inflammatory demyelinating polyneuropathies (CIDP)
  15. Detection of autoantibodies to the p200-epitope of SSA/Ro52 antigen. A comparison of two laboratory assays
  16. A new ELISA for autoantibodies to steroid 21-hydroxylase
  17. Antibodies to phosphatidylserine/prothrombin (aPS/PT) enhanced the diagnostic performance in Chinese patients with antiphospholipid syndrome
  18. Borderline positive antineutrophil cytoplasmic antibodies (ANCA)-PR3/MPO detection in a large cohort tertiary center: lessons learnt from a real-life experience
  19. Reference Values and Biological Variations
  20. Short- and medium-term biological variation estimates of red blood cell and reticulocyte parameters in healthy subjects
  21. Pediatric reference intervals for 1,25-dihydroxyvitamin D using the DiaSorin LIAISON XL assay in the healthy CALIPER cohort
  22. Reference intervals for biochemical, haemostatic and haematological parameters in healthy Chinese women during early and late pregnancy
  23. Cancer Diagnostics
  24. Use of circulating tumor cells in prospective clinical trials for NSCLC patients – standardization of the pre-analytical conditions
  25. High microRNA-28-5p expression in colorectal adenocarcinoma predicts short-term relapse of node-negative patients and poor overall survival of patients with non-metastatic disease
  26. Infectious Diseases
  27. Diagnosis of acute pediatric appendicitis from children with inflammatory diseases by combination of metabolic markers and inflammatory response variables
  28. Letters to the Editor
  29. Repeat requesting on hemolysed plasma potassium requests
  30. A case report of insulin autoimmune syndrome in a Central European individual
  31. Traceability of alkaline phosphatase measurement may also vary considerably using the same analytical system: the case of Abbott Architect
  32. Abiraterone acetate: a potential source of interference in testosterone assays
  33. Falsely elevated plasma testosterone concentrations in neonates: importance of LC-MS/MS measurements
  34. Rational approach to the primary evaluation of thyroid disease in paediatrics. Full thyroid profile vs. thyroid-stimulating hormone and free thyroxine only
  35. Diagnostic performance of the N-terminal pro-brain natriuretic peptide to detect an increased left atrial pressure in patients with persistent atrial fibrillation
  36. Is the DNA of placental origin packaged in exosomes isolated from plasma and serum of pregnant women?
  37. Gamma heavy chain disease evolving during the progression of chronic lymphocytic leukemia
  38. Congress Abstracts
  39. ISMD2018 Twelfth International Symposium on Molecular Diagnostics
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