TY - JOUR
T1 - Antibody-independent capture of circulating tumor cells of non-epithelial origin with the ApoStream® system
AU - Balasubramanian, Priya
AU - Kinders, Robert J.
AU - Kummar, Shivaani
AU - Gupta, Vishal
AU - Hasegawa, David
AU - Menachery, Anoop
AU - Lawrence, Scott M.
AU - Wang, Lihua
AU - Ferry-Galow, Katherine
AU - Davis, Darren
AU - Parchment, Ralph E.
AU - Tomaszewski, Joseph E.
AU - Doroshow, James H.
N1 - Funding Information:
The authors have read the journal's policy and the authors of this manuscript have the following competing interests: V.G., D.H., A.M., and D.D. are employees of ApoCell, Inc. P.B., R.J.K., S.M.L., L.W., K.F.G., and R.E.P. are affiliated with and received support in the form of salaries from The Frederick National Laboratory for Cancer Research, which is funded by the federal government and operated by Leidos Biomedical Research, Inc. Apostream is a trademark registered in the US Patent and Trademark Office. The ApoStream RUO device was developed by D.H., V.G., and A.M. (ApoCell, Inc.). The relevant patent for this instrument is entitled “Method and apparatus for isolation, capture and molecular analysis of target particle”; US patent application number: US20160209299A1. Data collection, data management and analysis were performed by The Frederick National Laboratory for Cancer Research. The specific roles of the authors are articulated in the ‘author contributions’ section. There are no further patents, products in development or marketed products to declare. This does not alter our adherence to all the PLOS ONE policies on sharing data and materials. All other authors have declared that no competing interests exist. ® ®
Publisher Copyright:
© This is an open access article, free of all copyright, and may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. The work is made available under the Creative Commons CC0 public domain dedication.
PY - 2017/4
Y1 - 2017/4
N2 - 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.
AB - 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.
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U2 - 10.1371/journal.pone.0175414
DO - 10.1371/journal.pone.0175414
M3 - Article
C2 - 28403214
AN - SCOPUS:85017436576
VL - 12
JO - PLoS One
JF - PLoS One
SN - 1932-6203
IS - 4
M1 - e0175414
ER -