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WIREs Nanomed Nanobiotechnol
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Nanobiotechnology for the capture and manipulation of circulating tumor cells

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Abstract A necessary step in metastasis is the dissemination of malignant cells into the bloodstream, where cancer cells travel throughout the body as circulating tumor cells (CTC) in search of an opportunity to seed a secondary tumor. CTC represent a valuable diagnostic tool: evidence indicates that the quantity of CTC in the blood has been shown to relate to the severity of the illness, and samples are readily obtained through routine blood draws. As such, there has been a push toward developing technologies to reliably detect CTC using a variety of molecular and immunocytochemical techniques. In addition to their use in diagnostics, CTC detection systems that isolate CTC in such a way that the cells remain viable will allow for the performance of live‐cell assays to facilitate the development of personalized cancer therapies. Moreover, techniques for the direct manipulation of CTC in circulation have been developed, intending to block metastasis in situ. We review a number of current and emerging micro‐ and nanobiotechnology approaches for the detection, capture, and manipulation of rare CTC aimed at advancing cancer treatment. WIREs Nanomed Nanobiotechnol 2012, 4:291–309. doi: 10.1002/wnan.168 This article is categorized under: Nanotechnology Approaches to Biology > Cells at the Nanoscale Implantable Materials and Surgical Technologies > Nanomaterials and Implants Therapeutic Approaches and Drug Discovery > Nanomedicine for Oncologic Disease

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In vitro/vivo manipulation of circulating tumor cells (CTC). (a) Schematic of the implantable shunt device for inducing apoptosis in CTC in vivo using E‐selectin for capture and tumor necrosis factor‐related apoptosis‐inducing ligand (TRAIL) to deliver the death signal. (b) HL60 leukemic cells are eliminated by approximately 30% following perfusion through the devices coated with both E‐selectin and TRAIL. ***P < 0.001. (c) Viability of normal mononuclear cells is not affected by the TRAIL coating, confirming the specificity of TRAIL for malignant cells. (d) Image showing the size of a compact shunt device for implantation (ruler units in cm). [(a–c) are reprinted with permission from Ref 87. Copyright 2008 Wiley Periodicals, Inc. and (d) is reprinted with permission from Ref 76. Copyright 2009 Elsevier Ltd]

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Halloysite nanotube coating enhances cell capture in the selectin‐functionalized device. (a) Schematic shows how the orientation of nanotubes within the surface coating promotes enhanced cell capture by presenting selectin molecules into the flow. (b) Atomic force microscopy shows that halloysite nanotubes extend above the surface. (c) P‐selectin adsorption is increased on the nanotube coating over a range of P‐selectin incubating solution concentrations. (d) The number of cells captured in the selectin‐functionalized device as a function of inner surface area is significantly enhanced on the nanotube coating over a range of P‐selectin concentrations. Representative micrographs of the control device (e) and nanotube‐coated device (f) show that the number of cells captured is visibly increased. ***P < 0.001. (Figures are reprinted with permission from Ref 79. Copyright 2010 American Chemical Society)

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Silica (SiO2) nanoparticle (10–15 nm) coatings augment cell capture in a selectin‐functionalized device. (a) Schematic of nanoparticle coating shows the impact on P‐selectin adsorption. (b) Silica nanoparticles were coated onto the device surface using two different adhesives and P‐selectin adsorption was significantly enhanced using both methods. Adsorption quantification was determined using fluorescent antibodies against P‐selectin. (c) The number of cells captured was greater using the silica nanoparticle coating over the physiological range of shear stresses compared to the smooth surface. *P < 0.05; **P > 0.05. (Figures are reprinted with permission from Ref 77. Copyright 2010 American Chemical Society)

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Circulating tumor cells (CTC) capture from blood in a selectin‐functionalized microtube. (a) Various numbers of KG1a acute myeloid leukemia cells were spiked into blood. Samples were perfused through MicroRenathane (MRE) tubes coated with either 50 µg/mL anti‐CD34 alone or coated with both 50 µg/mL anti‐CD34 and 0.5 µg/mL P‐selectin. Approximately 50% of KG1a cells spiked into blood samples were recovered on the bimolecular‐coated MRE tube. (b) Representative low‐intensity bright field micrograph of cells captured in a bimolecular‐coated MRE tube. (c) Fluorescent KG1a cell captured in bimolecular‐coated MRE tube. Eighty‐four fluorescently labeled KG1a cells were spiked into 4 mL blood. Scale bars are 20 µm. Arrow indicatesa positive KG1a adhesion event. (Data are adapted and micrographs are reprinted with permission from Ref 76. Copyright 2009 Elsevier Ltd)

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Schematic describing the utility of the three major classes (a–c) of technologies for capture and/or manipulation of circulating tumor cells.

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Efficient delivery of small‐interfering RNA (siRNA) using P‐selectin targeted liposomes coated in an implantable shunt device. (a) Liposomes are encapsulated with siRNA and covalently coated with P‐selectin and polyethylene glycol (PEG). (b) Scanning electron microscopy shows the size and shape of the encapsulated liposomes. The average diameter was found to be approximately 150 nm. (c) Red fluorescent RNA encapsulated within the liposomes was efficiently delivered to HL60 cells perfused through the liposome‐coated shunt device. (d) Significant ELA2 gene knockdown is shown using real‐time quantitative polymerase chain reaction (PCR) to indicate the effectiveness of this siRNA delivery technique. **P < 0.01. (Figures are reproduced with permission from Ref 92. Copyright 2009 Macmillan Publishers Ltd)

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Therapeutic Approaches and Drug Discovery > Nanomedicine for Oncologic Disease
Implantable Materials and Surgical Technologies > Nanomaterials and Implants
Nanotechnology Approaches to Biology > Cells at the Nanoscale

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