Microsieves for the detection of circulating tumor cells in leukapheresis product in non-small cell lung cancer patients

Menno Tamminga, Lisa Oomens, T. Jeroen N. Hiltermann, Kiki C. Andree, Arjan Tibbe, Joska Broekmaat, Ed Schuuring, Leon W.M.M. Terstappen, Harry J.M. Groen*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

2 Citations (Scopus)
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Abstract

Background: Circulating tumor cells (CTC) in non-small cell lung cancer (NSCLC) patients are a prognostic and possible therapeutic marker, but have a low frequency of appearance. Diagnostic leukapheresis (DLA) concentrates CTC and mononuclear cells from the blood. We evaluated a protocol using two VyCAP microsieves to filter DLA product of NSCLC patients and enumerate CTC, compared with CellSearch as a gold standard. Methods: DLA was performed in NSCLC patients before starting treatment. DLA product equaling 2×108 leukocytes was diluted to 9 mL with CellSearch dilution buffer in a Transfix CTC tube. Within 72 hours the sample was filtered with a 7 μm pore microsieve and subsequently over a 5μm pore microsieve. CTC were defined as nucleated cells which stained for cytokeratin, but lacked CD45 and CD16. CellSearch detected CTC in the same volume of DLA. Results: Of 29 patients a median of 1.4 mL DLA product (range, 0.5-4.1) was filtered (2% of total product) successfully in 93% and 45% of patients using 7 and 5 μm pores, respectively. Two DLA products were unevaluable for CTC detection. Clogging of the 5 μm but not 7 μm microsieves was positively correlated with fixation time (ρ=0.51, P<0.01). VyCAP detected CTC in 44% (12/27) of DLA products. Median CTC count per mL DLA was 0 [interquartile range (IQR): 0-1]. CellSearch detected CTC in 63% of DLA products (median =0.9 CTC per mL DLA, IQR: 0-2.1). CTC counts detected by CellSearch were significantly higher compared with VyCAP (P=0.05). Conclusions: VyCAP microsieves can identify CTC in DLA product, but workflows need to be optimized.

Original languageEnglish
Pages (from-to)1093-1100
Number of pages8
JournalTranslational Lung Cancer Research
Volume9
Issue number4
DOIs
Publication statusPublished - Aug 2020

Keywords

  • Biomarker
  • Circulating tumor cell (CTC)
  • Diagnostic leukapheresis (DLA)
  • Liquid biopsy
  • Non-small lung cancer (NSCLC)
  • VyCAP microsieves

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