TY - JOUR
T1 - Microsieves for the detection of circulating tumor cells in leukapheresis product in non-small cell lung cancer patients
AU - Tamminga, Menno
AU - Oomens, Lisa
AU - Hiltermann, T. Jeroen N.
AU - Andree, Kiki C.
AU - Tibbe, Arjan
AU - Broekmaat, Joska
AU - Schuuring, Ed
AU - Terstappen, Leon W.M.M.
AU - Groen, Harry J.M.
PY - 2020/8
Y1 - 2020/8
N2 - 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.
AB - 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.
KW - Biomarker
KW - Circulating tumor cell (CTC)
KW - Diagnostic leukapheresis (DLA)
KW - Liquid biopsy
KW - Non-small lung cancer (NSCLC)
KW - VyCAP microsieves
UR - http://www.scopus.com/inward/record.url?scp=85084849016&partnerID=8YFLogxK
U2 - 10.21037/tlcr-19-413
DO - 10.21037/tlcr-19-413
M3 - Article
AN - SCOPUS:85084849016
SN - 2218-6751
VL - 9
SP - 1093
EP - 1100
JO - Translational Lung Cancer Research
JF - Translational Lung Cancer Research
IS - 4
ER -
