Antibody-independent capture of circulating tumor cells of non-epithelial origin with the ApoStream® system

PLoS One. 2017 Apr 12;12(4):e0175414. doi: 10.1371/journal.pone.0175414. eCollection 2017.

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.

MeSH terms

  • A549 Cells
  • Biomarkers, Tumor / metabolism
  • Case-Control Studies
  • Cell Separation / instrumentation*
  • Cell Separation / methods
  • Co-Repressor Proteins
  • Humans
  • Neoplastic Cells, Circulating / metabolism*
  • Repressor Proteins / metabolism
  • Sarcoma, Alveolar Soft Part / blood*
  • Sarcoma, Alveolar Soft Part / pathology

Substances

  • Biomarkers, Tumor
  • Co-Repressor Proteins
  • Repressor Proteins
  • TLE1 protein, human

Grant support

This project has been funded in whole or in part with federal funds from the National Cancer Institute, National Institutes of Health, under Contract No. HHSN261200800001E. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. Government. The Frederick National Laboratory for Cancer Research, which is funded by the federal government and operated by Leidos Biomedical Research, Inc., provided support in the form of salaries for authors P.B., R.J.K., S.M.L., L.W., K.F.G., and R.E.P., but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. Leidos Biomedical Research, Inc. is the prime contractor for the operation of Frederick National Laboratory and is therefore precluded from conducting any business that represents a conflict of interest with this contract. ApoCell, Inc provided support in the form of salaries for authors V.G., D.H., A.M., and D.D, but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of all authors are articulated in the 'author contributions' section.