A novel method for the in vivo isolation of circulating tumor cells from peripheral blood of cancer patients using a functionalized and structured medical wire

Abstract

The isolation of circulating tumor cells (CTCs) from the blood of patients afflicted with solid malignant tumors becomes increasingly important as it may serve as a ‘liquid biopsy’ with the potential of monitoring the course of the cancer disease and its response to cancer therapy, with subsequent molecular characterization. For this purpose, we functionalized a structured medical Seldinger guidewire (FSMW), normally used to obtain safe access to blood vessels and other organ cavities, with a chimeric monoclonal antibody directed to the cell surface expressed epithelial cell surface adhesion molecule (EpCAM). This medical device was optimized in vitro and its biocompatibility was tested according to the regulations for medical devices and found to be safe with no noteworthy side effects. Suitability, specificity and sensitivity of the FSMW to catch and enrich CTCs in vivo from circulating peripheral blood were tested in 24 breast cancer or non-small cell lung cancer (NSCLC) patients and in 29 healthy volunteers. For this, the FSMW was inserted through a standard venous cannula into the cubital veins of healthy volunteers or cancer patients for the duration of 30 min. After removal, CTCs were identified by immunocytochemical staining of EpCAM and/or cytokeratins and staining of their nuclei and counted. The FSMW successfully enriched EpCAM-positive CTCs from 22 of the 24 patients, with a median of 5.5 (0–50) CTCs in breast cancer (n=12) and 16 (2–515) CTCs in NSCLC (n=12). CTCs could be isolated across all tumor stages, including early stage cancer, in which distant metastases were not yet diagnosed, while no CTCs could be detected in healthy volunteers. In this observatory study, no adverse effects were noted. Evidently, the FSMW has the potential to become an important device to enrich CTCs in vivo for monitoring the course of the cancer disease and the efficacy of anticancer treatment.

Improving pancreatic cancer diagnosis using circulating tumor cells: prospects for staging and single-cell analysis

“…The various methods also result in vastly different numbers of detected CTCs. For example, the CellCollector in vivo CTC platform detected between 1 and 5 CTCs in 1.5 l of blood versus 12 and 3167 CTCs/ml of blood reported using the CTC-Chip [41,42]. …”

Circulating tumor cell technologies

“…The antibodies used for selection are typically tethered to either the device surface or a magnetic substance (i.e., (Miltenyi et al, 1990;Pluim et al, 2012) MagSweeper EpCAM High purity, can process WB, 9 mL/h throughput Kim et al, 2014;Lohr et al, 2014;Powell et al, 2012;Talasaz et al, 2009 (Harb et al, 2013). mF, IM CTC-iChip EpCAM, Size Pos/neg enrichment, size-based separation debulks WB, inertial focusing aids in magnetic deflection, 8 mL/h (Karabacak et al, 2014;Ozkumur et al, 2013) IM, in vivo GILUPI CellCollectorä EpCAM Can process large volumes of blood (Saucedo-Zeni et al, 2012) Immunoaffinity e Negative Enrichment Antibodies targeting leukocyte-associated antigens are tethered to magnetic particles or the device surface deplete unwanted background cells.…”

Circulating tumor cells and miRNAs as prognostic markers in neuroendocrine neoplasms

“…A possible efficient strategy to overcome this problem is to directly target the cells in vivo. GILUPI GmbH approach is based on the application of an antibodycoated wire into a peripheral arm vein for a duration of 30 min (Saucedo-Zeni et al 2012). After removing the wire, isolated CTCs can be removed for immunocytochemical and molecular analysis.…”