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

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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 pages10
JournalTranslational lung cancer research
Volume9
Issue number4
DOIs
Publication statusPublished - Aug-2020

Keywords

  • Diagnostic leukapheresis (DLA)
  • circulating tumor cell (CTC)
  • non-small lung cancer (NSCLC)
  • liquid biopsy
  • biomarker
  • VyCAP microsieves
  • DIAGNOSTIC LEUKAPHERESIS

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