Abstract
Abstract Circulating tumor cell (CTC) has recently grabbed numerous headlines with the promise for personalized cancer diagnostics and treatment. Its clinical value, in terms of presence and the number, has been demonstrated in accumulated results for prognosis of cancer progression and mortality, and prediction of therapy response. CTCs, the leukemic phase of solid tumor, further give us the hope for a rapid and affordable optical imaging tool as CTCs are readily accessible and optically observable. Fluorescence based cell imaging has been widely spread to detect CTCs. However, it is still a challenge to accomplish a point-of-care test (POCT) due to many limitations such as the cost, fluorescence stability, variations in fluorescence detection and image processing, and data interpretation. Chromogenic labeling may offer exciting perspectives in POCT for CTC detection along with the application of metallic signal enhancement. Based on this concept, TeloVISION LLC developed a palm-sized, manually operated, POCT device for CTC detection. The device, which bears a membrane with rod-shaped, micron-sized pores, processes the whole blood without vigorous manipulation. By manually rotating the device counter-clockwise, the entire CTC assay is split into 7 steps, performed inside the device, and completed in 40 minutes. To validate the device, whole blood was withdrawn from the animals bearing metastatic tumors. Due to the restriction of the membrane, CTCs larger than 5µm size were retained while majority of the normal cells were removed due to the discrepancy in deformability. In addition, the filtration was driven by gravity so that the mechanical damage from external force was avoided. After filtration, enriched CTCs were identified by DAB stain in the presence of horseradish peroxidase (HRP) tagged anti-cytokeratin (CK) and the signal was intensified 50-fold by metal precipitation. CTCs were found in blood as well as the circulating tumor microemboli (CTM) by both chromogenic labeling and fluorescence staining (control). Heterogeneity in the CK expression was seen for both CTCs and CTM. The recovery of the device was 94% compared to that by hemocytometer. We conclude that chromogenic labeling with the signal enhancement by in situ metal precipitation can provide high-resolution, sensitive CTC detection, and prevent the false positive results as seen in fluorescent labeling due to autofluorescence and fluorescence ghost in the blood. This new portable device may provide an efficient low-cost CTC detection in clinical settings. Adjustment of the antibodies will expand the field of application such as sarcoma and melanoma, non-epithelial derived CTCs. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 4059. doi:1538-7445.AM2012-4059
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