Versatile immunomagnetic nanocarrier platform for capturing cancer cells.

Sensitive and quantitative assessment of changes in circulating tumor cells (CTCs) can help in cancer prognosis and in the evaluation of therapeutics efficacy. However, extremely low occurrence of CTCs in the peripheral blood (approximately one CTC per billion blood cells) and potential changes in molecular biomarkers during the process of epithelial to mesenchymal transition create technical hurdles to the enrichment and enumeration of CTCs. Recently, efforts have been directed toward development of antibody-capture assays based on the expression of the common biomarker-the epithelial cell adhesion molecule (EpCAM) of epithelium-derived cancer cells. Despite some promising results, the assays relying on EpCAM capture have shown inconsistent sensitivity in clinical settings and often fail to detect CTCs in patients with metastatic cancer. We have addressed this problem by the development of an assay based on hybrid magnetic/plasmonic nanocarriers and a microfluidic channel. In this assay, cancer cells are specifically targeted by antibody-conjugated magnetic nanocarriers and are separated from normal blood cells by a magnetic force in a microfluidic chamber. Subsequently, immunofluorescence staining is used to differentiate CTCs from normal blood cells. We demonstrated in cell models of colon, breast, and skin cancers that this platform can be easily adapted to a variety of biomarkers, targeting both surface receptor molecules and intracellular biomarkers of epithelial-derived cancer cells. Experiments in whole blood showed capture efficiency greater than 90% when two cancer biomarkers are used for cell capture. Thus, the combination of immunotargeted magnetic nanocarriers with microfluidics provides an important platform that can improve the effectiveness of current CTC assays by overcoming the problem of heterogeneity of tumor cells in the circulation.

[1]  Stephan Barcikowski,et al.  Advanced nanoparticle generation and excitation by lasers in liquids. , 2013, Physical chemistry chemical physics : PCCP.

[2]  Rebecca Richards-Kortum,et al.  Plasmonic nanosensors for imaging intracellular biomarkers in live cells. , 2007, Nano letters.

[3]  Hong Yang,et al.  “Pulling” Nanoparticles into Water: Phase Transfer of Oleic Acid Stabilized Monodisperse Nanoparticles into Aqueous Solutions of α-Cyclodextrin , 2003 .

[4]  J Aaron,et al.  Directional conjugation of antibodies to nanoparticles for synthesis of multiplexed optical contrast agents with both delivery and targeting moieties , 2008, Nature Protocols.

[5]  J. Ostrander,et al.  Monitoring of receptor dimerization using plasmonic coupling of gold nanoparticles. , 2011, ACS nano.

[6]  Mieke Schutte,et al.  Detection of circulating tumor cells in breast cancer may improve through enrichment with anti-CD146 , 2011, Breast Cancer Research and Treatment.

[7]  W. Parak,et al.  Fluorescent, magnetic and plasmonic—Hybrid multifunctional colloidal nano objects , 2012 .

[8]  Alison Stopeck,et al.  Circulating tumor cells, disease progression, and survival in metastatic breast cancer. , 2004, The New England journal of medicine.

[9]  B. Reinhard,et al.  Quantification of differential ErbB1 and ErbB2 cell surface expression and spatial nanoclustering through plasmon coupling. , 2012, Nano letters.

[10]  Konstantin V Sokolov,et al.  Hybrid plasmonic magnetic nanoparticles as molecular specific agents for MRI/optical imaging and photothermal therapy of cancer cells , 2007 .

[11]  Q. Wei,et al.  Plasmon-resonant nanoparticles and nanostars with magnetic cores: synthesis and magnetomotive imaging. , 2010, ACS nano.

[12]  J. Schellens,et al.  Circulating tumor cells as pharmacodynamic biomarker in early clinical oncological trials. , 2011, Cancer treatment reviews.

[13]  J. Bobbitt Periodate oxidation of carbohydrates. , 1956, Advances in carbohydrate chemistry.

[14]  Jeffrey N. Anker,et al.  Biosensing with plasmonic nanosensors. , 2008, Nature materials.

[15]  Alison Stopeck,et al.  Circulating tumor cells: a novel prognostic factor for newly diagnosed metastatic breast cancer. , 2005, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[16]  Kazunori Hoshino,et al.  Microchip-based immunomagnetic detection of circulating tumor cells. , 2011, Lab on a chip.

[17]  M. Bissell A First-Generation Multiplex Biomarker Analysis of Urine For The Early Detection of Prostate Cancer , 2009 .

[18]  S. Digumarthy,et al.  Isolation of rare circulating tumour cells in cancer patients by microchip technology , 2007, Nature.

[19]  Brigitte Rack,et al.  Detection of Circulating Tumor Cells in Peripheral Blood of Patients with Metastatic Breast Cancer: A Validation Study of the CellSearch System , 2007, Clinical Cancer Research.

[20]  Jin Luo,et al.  Monodispersed core-shell Fe3O4@Au nanoparticles. , 2005, The journal of physical chemistry. B.

[21]  R. Chadwick,et al.  EGFR and EGFRvIII Expression in Primary Breast Cancer and Cell Lines , 2004, Breast Cancer Research and Treatment.

[22]  Ru-Fang Yeh,et al.  Molecular Biomarker Analyses Using Circulating Tumor Cells , 2010, PloS one.

[23]  Mieke Schutte,et al.  Anti-Epithelial Cell Adhesion Molecule Antibodies and the Detection of Circulating Normal-Like Breast Tumor Cells , 2009, Journal of the National Cancer Institute.

[24]  Sanjay Shete,et al.  uPAR and HER-2 gene status in individual breast cancer cells from blood and tissues , 2006, Proceedings of the National Academy of Sciences.

[25]  Kort Travis,et al.  Plasmon resonance coupling of metal nanoparticles for molecular imaging of carcinogenesis in vivo. , 2007, Journal of biomedical optics.

[26]  Tanja Fehm,et al.  Stem cell and epithelial-mesenchymal transition markers are frequently overexpressed in circulating tumor cells of metastatic breast cancer patients , 2009, Breast Cancer Research.

[27]  A. P. Leonov,et al.  Gyromagnetic imaging: dynamic optical contrast using gold nanostars with magnetic cores. , 2009, Journal of the American Chemical Society.

[28]  J. Bono,et al.  All circulating EpCAM+CK+CD45- objects predict overall survival in castration-resistant prostate cancer. , 2010, Annals of oncology : official journal of the European Society for Medical Oncology.

[29]  B. Reinhard,et al.  Illuminating epidermal growth factor receptor densities on filopodia through plasmon coupling. , 2011, ACS nano.

[30]  David Elashoff,et al.  Salivary transcriptomic biomarkers for detection of resectable pancreatic cancer. , 2010, Gastroenterology.

[31]  Kort Travis,et al.  Dynamic imaging of molecular assemblies in live cells based on nanoparticle plasmon resonance coupling. , 2009, Nano letters.

[32]  Robert Langer,et al.  Impact of nanotechnology on drug delivery. , 2009, ACS nano.

[33]  Tuo Wei,et al.  Size-dependent localization and penetration of ultrasmall gold nanoparticles in cancer cells, multicellular spheroids, and tumors in vivo. , 2012, ACS nano.

[34]  Do Kyung Kim,et al.  Antibiofouling polymer-coated superparamagnetic iron oxide nanoparticles as potential magnetic resonance contrast agents for in vivo cancer imaging. , 2006, Journal of the American Chemical Society.

[35]  Peng Chen,et al.  Computational analysis of microfluidic immunomagnetic rare cell separation from a particulate blood flow. , 2012, Analytical chemistry.