title: MINDEC-An Enhanced Negative Depletion Strategy for Circulating Tumour Cell Enrichment.
abstract: Most current methods of circulating tumour cell (CTC) enrichment target the epithelial protein EpCAM, which is commonly expressed in adenocarcinoma cells. However, such methods will not recover the fraction of CTCs that have a non-epithelial phenotype due to epithelial-mesenchymal transition. For phenotype-independent CTC enrichment, we developed a new enhanced negative depletion strategy-termed MINDEC-that is based on multi-marker (CD45, CD16, CD19, CD163, and CD235a/GYPA) depletion of blood cells rather than targeted enrichment of CTCs. Here we validated the performance of MINDEC using epithelial and mesenchymal cancer cell lines, demonstrating a mean recovery of 82 ± 10%, high depletion (437 ± 350 residual white blood cells (WBCs)/mL peripheral blood), linearity between spiked and recovered cells (correlation coefficient: r = 0.995), and a low detection limit (≥1 cell recovered in all four replicates spiked with 3 cells). For clinical validation of this method, we enumerated CTCs in peripheral blood samples from patients with metastatic pancreatic cancer, detecting CTCs in 15 of 21 blood samples (71%) from 9 patients. The promising performance of the MINDEC enrichment strategy in our study encourages validation in larger clinical trials.
authors.affiliation: Department of Haematology and Oncology, Stavanger University Hospital, N-4068 Stavanger, Norway.|Laboratory for Molecular Biology, Stavanger University Hospital, N-4068 Stavanger, Norway.|Department of Mathematics and Natural Sciences, University of Stavanger, N-4036 Stavanger, Norway.
authors.forename: Morten
authors.initials: M
authors.lastname: Lapin
authors.affiliation: Department of Haematology and Oncology, Stavanger University Hospital, N-4068 Stavanger, Norway.|Laboratory for Molecular Biology, Stavanger University Hospital, N-4068 Stavanger, Norway.
authors.forename: Kjersti
authors.initials: K
authors.lastname: Tjensvoll
authors.affiliation: Department of Haematology and Oncology, Stavanger University Hospital, N-4068 Stavanger, Norway.|Laboratory for Molecular Biology, Stavanger University Hospital, N-4068 Stavanger, Norway.
authors.forename: Satu
authors.initials: S
authors.lastname: Oltedal
authors.affiliation: Department of Haematology and Oncology, Stavanger University Hospital, N-4068 Stavanger, Norway.
authors.forename: Tove
authors.initials: T
authors.lastname: Buhl
authors.affiliation: Department of Haematology and Oncology, Stavanger University Hospital, N-4068 Stavanger, Norway.
authors.forename: Bjørnar
authors.initials: B
authors.lastname: Gilje
authors.affiliation: Department of Haematology and Oncology, Stavanger University Hospital, N-4068 Stavanger, Norway.|Laboratory for Molecular Biology, Stavanger University Hospital, N-4068 Stavanger, Norway.
authors.forename: Rune
authors.initials: R
authors.lastname: Smaaland
authors.affiliation: Department of Haematology and Oncology, Stavanger University Hospital, N-4068 Stavanger, Norway.|Laboratory for Molecular Biology, Stavanger University Hospital, N-4068 Stavanger, Norway.
authors.forename: Oddmund
authors.initials: O
authors.lastname: Nordgård
mesh_terms: D015703:Antigens, CD / Q000097:blood; D018941:Antigens, CD19 / Q000097:blood; D015214:Antigens, Differentiation, Myelomonocytic / Q000097:blood; D000951:Antigens, Neoplasm / Q000097:blood*; D014408:Biomarkers, Tumor / Q000097:blood; D001943:Breast Neoplasms / Q000097:blood; D015815:Cell Adhesion Molecules / Q000097:blood*; D045744:Cell Line, Tumor; D002469:Cell Separation / Q000379:methods; D003584:Cytological Techniques; D000071858:Epithelial Cell Adhesion Molecule / Q000378:metabolism*; D058750:Epithelial-Mesenchymal Transition; D005260:Female; D058851:GPI-Linked Proteins / Q000097:blood; D006021:Glycophorins / Q000097:blood; D006801:Humans; D017493:Leukocyte Common Antigens / Q000097:blood; D057230:Limit of Detection; D008175:Lung Neoplasms / Q000097:blood; D061986:MCF-7 Cells; D008654:Mesothelioma / Q000097:blood; D009360:Neoplastic Cells, Circulating / Q000473:pathology*; D010190:Pancreatic Neoplasms / Q000097:blood; D010641:Phenotype; D011956:Receptors, Cell Surface / Q000097:blood; D017452:Receptors, IgG / Q000097:blood; D015203:Reproducibility of Results; D000098902:CD163 Antigen
chemical_list: D015703:Antigens, CD; D018941:Antigens, CD19; D015214:Antigens, Differentiation, Myelomonocytic; D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D000098902:CD163 Antigen; C000604811:CD19 molecule, human; D015815:Cell Adhesion Molecules; C499379:EPCAM protein, human; D000071858:Epithelial Cell Adhesion Molecule; C428200:FCGR3B protein, human; D058851:GPI-Linked Proteins; C409972:GYPA protein, human; D006021:Glycophorins; D011956:Receptors, Cell Surface; D017452:Receptors, IgG; D017493:Leukocyte Common Antigens; C516769:PTPRC protein, human