title: Antibody-independent capture of circulating tumor cells of non-epithelial origin with the ApoStream® system.
abstract: Circulating tumor cells (CTCs) are increasingly employed for research and clinical monitoring of cancer, though most current methods do not permit the isolation of non-epithelial tumor cells. Furthermore, CTCs isolated with antibody-dependent methods are not suitable for downstream experimental uses, including in vitro culturing and implantation in vivo. In the present study, we describe the development, validation, and transfer across laboratories of a new antibody-independent device for the enrichment of CTCs from blood samples of patients with various cancer diagnoses. The ApoStream® device uses dielectrophoresis (DEP) field-flow assist to separate non-hematopoietic cells from the peripheral blood mononuclear fraction by exposing cells in a laminar flow stream to a critical alternating current frequency. The ApoStream® device was calibrated and validated in a formal cross-laboratory protocol using 3 different cancer cell lines spanning a range of distinct phenotypes (A549, MDA-MB-231, and ASPS-1). In spike-recovery experiments, cancer cell recovery efficiencies appeared independent of spiking level and averaged between 68% and 55%, depending on the cell line. No inter-run carryover was detected in control samples. Moreover, the clinical-readiness of the device in the context of non-epithelial cancers was evaluated with blood specimens from fifteen patients with metastatic sarcoma. The ApoStream® device successfully isolated CTCs from all patients with sarcomas examined, and the phenotypic heterogeneity of the enriched cells was demonstrated by fluorescence in situ hybridization or with multiplex immunophenotyping panels. Therefore, the ApoStream® technology expands the clinical utility of CTC evaluation to mesenchymal cancers.
authors.affiliation: Laboratory of Human Toxicology and Pharmacology, Applied/ Developmental Research Support Directorate, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, Frederick, Maryland, United States of America.
authors.forename: Priya
authors.initials: P
authors.lastname: Balasubramanian
authors.affiliation: Laboratory of Human Toxicology and Pharmacology, Applied/ Developmental Research Support Directorate, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, Frederick, Maryland, United States of America.
authors.forename: Robert J
authors.identifier: http://orcid.org/0000-0002-5310-3367
authors.initials: RJ
authors.lastname: Kinders
authors.affiliation: Division of Cancer Treatment and Diagnosis, National Cancer Institute, Bethesda, Maryland, United States of America.
authors.forename: Shivaani
authors.initials: S
authors.lastname: Kummar
authors.affiliation: ApoCell, Inc., Houston, Texas, United States of America.
authors.forename: Vishal
authors.initials: V
authors.lastname: Gupta
authors.affiliation: ApoCell, Inc., Houston, Texas, United States of America.
authors.forename: David
authors.initials: D
authors.lastname: Hasegawa
authors.affiliation: ApoCell, Inc., Houston, Texas, United States of America.
authors.forename: Anoop
authors.initials: A
authors.lastname: Menachery
authors.affiliation: Laboratory of Human Toxicology and Pharmacology, Applied/ Developmental Research Support Directorate, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, Frederick, Maryland, United States of America.
authors.forename: Scott M
authors.initials: SM
authors.lastname: Lawrence
authors.affiliation: Laboratory of Human Toxicology and Pharmacology, Applied/ Developmental Research Support Directorate, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, Frederick, Maryland, United States of America.
authors.forename: Lihua
authors.initials: L
authors.lastname: Wang
authors.affiliation: Laboratory of Human Toxicology and Pharmacology, Applied/ Developmental Research Support Directorate, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, Frederick, Maryland, United States of America.
authors.forename: Katherine
authors.initials: K
authors.lastname: Ferry-Galow
authors.affiliation: ApoCell, Inc., Houston, Texas, United States of America.
authors.forename: Darren
authors.initials: D
authors.lastname: Davis
authors.affiliation: Laboratory of Human Toxicology and Pharmacology, Applied/ Developmental Research Support Directorate, Leidos Biomedical Research, Inc., Frederick National Laboratory for Cancer Research, Frederick, Maryland, United States of America.
authors.forename: Ralph E
authors.initials: RE
authors.lastname: Parchment
authors.affiliation: Division of Cancer Treatment and Diagnosis, National Cancer Institute, Bethesda, Maryland, United States of America.
authors.forename: Joseph E
authors.initials: JE
authors.lastname: Tomaszewski
authors.affiliation: Division of Cancer Treatment and Diagnosis, National Cancer Institute, Bethesda, Maryland, United States of America.
authors.forename: James H
authors.initials: JH
authors.lastname: Doroshow
mesh_terms: D000072283:A549 Cells; D014408:Biomarkers, Tumor / Q000378:metabolism; D016022:Case-Control Studies; D002469:Cell Separation / Q000295:instrumentation* / Q000379:methods; D056970:Co-Repressor Proteins; D006801:Humans; D009360:Neoplastic Cells, Circulating / Q000378:metabolism*; D012097:Repressor Proteins / Q000378:metabolism; D018234:Sarcoma, Alveolar Soft Part / Q000097:blood* / Q000473:pathology
chemical_list: D014408:Biomarkers, Tumor; D056970:Co-Repressor Proteins; D012097:Repressor Proteins; C499043:TLE1 protein, human