Biomolecular Surfaces for the Capture and Reprogramming of Circulating Tumor Cells

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Circulating Tumor Cells(CTC)have the potential to be used clinically as a diagnostic tool and a treatment tool in the fieldof oncology.As a diagnostic tool,CTC may be used to indicate the presence of a tumor before it is large enough to cause noticeablesymptoms.As a treatment tool,CTC isolated from patients may be used to test the efficacy of chemotherapy options topersonalize patient treatment.One way for tumors to spread is through metastasis via the circulatory system.CTC are able toexploit the natural leukocyte recruitment process that is initially mediated by rolling on transient selectin bonds.Our capturedevices take advantage of this naturally occurring recruitment step to isolate CTC from whole blood by flowing samples throughselectin and antibody-coated microtubes.Whole blood was spiked with a known concentration of labeled cancer cells and thenperfused through pre-coated microtubes.Microtubes were then rinsed to remove unbound cells and the number of labeled cellscaptured on the lumen was assessed.CTC were successfully captured from whole blood at a clinically relevant level on the orderof 10 cells per mL.Combination tubes with selectin and antibody coated surface exhibited higher capture rate than tubes coatedwith selectin alone or antibody alone.Additionally,CTC capture was demonstrated with the KG 1 a hematopoietic cell line andthe DU 145 epithelial cell line.Thus,the in vivo process of selectin-mediated CTC recruitment to distant vessel walls can be usedin vitro to target CTC to a tube lumen.The biomolecular coatings can also be used to capture CTC of hematopoietic andepithelial tumor origin and is demonstrated to sensitivities down to the order of 10 CTC per mL.In a related study aimed at reducing the blood borne metastatic cancer load,we have shown that cells captured to a surfacecan be neutralized by a receptor-mediated biochemical signal.In the proposed method we have shown that using a combinedselectin and TRAIL(TNF Related Apoptosis Inducing Ligand or Apo 2L)functionalized surface we are able to kill about 30%of the captured cells in a short duration of one hour whereas it took about 4 hours to kill the same proportion of cells withoutflow on a similarly functionalized surface.Here we have taken the approach a step further by showing that with very small dosesof chemotherapeutic agents like bortezomib,we can increase the kill rate of CTC,thus allowing the device to function in scenarioswhere the patient is undergoing treatment.We show here that,with leukemic cells treated with bortezomib we are able tokill about 41% of the captured cells. Circulating Tumor Cells (CTCs) have the potential to be used clinically as a diagnostic tool and a treatment tool in the field of oncology. As a diagnostic tool, CTC may be used to indicate the presence of a tumor before it is large enough to cause noticeable symptoms .As a treatment tool, CTC isolated from patients may be used to test the efficacy of chemotherapy options topersonalize patient treatment. One way for tumors to spread is through metastasis via the circulatory system. CTC are able to expit the natural leukocyte recruitment process that is initially mediated by rolling on transient selectin bonds. Our capture happenvices take advantage of this naturally occurring recruitment step to isolate CTC from whole blood by flowing samples throughselectin and antibody-coated microtubes. Well blood was spiked with a known concentration of labeled cancer cells and thenperfused through pre -coated microtubes. Microtubes were then rinsed to remove unbound cells and the number of labeled cells captured on the lumen was assessed. CTC were successfully captured from whole blood at a clinically relevant level on the order of 10 cells per mL. Combines tubes with selectin and antibody coated surface exhibit higher capture rate than tubes coated with selectin alone or antibody alone. Additionally, CTC capture was demonstrated with the KG 1 a hematopoietic cell line and the DU 145 epithelial cell line. Thus, the in vivo process of selectin-mediated CTC recruitment to distant vessel walls can be used in vitro to target CTC to a tube lumen. biomolecular coatings can also be used to capture CTC of hematopoietic andepithelial tumor origin and is demonstrated to sensitivities down to the order of 10 CTC per mL. a related study aimed at reducing the blood borne metastatic cancer load, we have shown that cells captured to a surfacecan be neutralized by a receptor-mediated biochemical signal. In the proposed method we have shown that using a combination selectin and TRAIL (TNF Related Apoptosis Inducing Ligand or or Apo 2L) functionalized surface we are able able kill kill about 30% of the captured cells in a short duration of one hour it it took about 4 hours to kill the same proportion of cells withoutflow on a similarly functionalized surface. the approach a step further by showing that with very small doses of chemotherapeutic agents like bortezomib, we can increase the kill rate of CTC, thus allowing the device to function in scenarios where the patient is undergoing treatment. We show here that, with leukemic cells treated with bortezomib we are able tokill about 41% of the captured cells.
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