Abstract
BackgroundCirculating tumor cells (CTCs) are cancer cells that can be isolated via liquid biopsy from blood and can be phenotypically and genetically characterized to provide critical information for guiding cancer treatment. Current analysis of CTCs is hindered by the throughput, selectivity and specificity of devices or assays used in CTC detection and isolation.Methodology/Principal FindingsHere, we enriched and characterized putative CTCs from blood samples of patients with both advanced stage metastatic breast and lung cancers using a novel multiplexed spiral microfluidic chip. This system detected putative CTCs under high sensitivity (100%, n = 56) (Breast cancer samples: 12–1275 CTCs/ml; Lung cancer samples: 10–1535 CTCs/ml) rapidly from clinically relevant blood volumes (7.5 ml under 5 min). Blood samples were completely separated into plasma, CTCs and PBMCs components and each fraction were characterized with immunophenotyping (Pan-cytokeratin/CD45, CD44/CD24, EpCAM), fluorescence in-situ hybridization (FISH) (EML4-ALK) or targeted somatic mutation analysis. We used an ultra-sensitive mass spectrometry based system to highlight the presence of an EGFR-activating mutation in both isolated CTCs and plasma cell-free DNA (cf-DNA), and demonstrate concordance with the original tumor-biopsy samples.Conclusions/SignificanceWe have clinically validated our multiplexed microfluidic chip for the ultra high-throughput, low-cost and label-free enrichment of CTCs. Retrieved cells were unlabeled and viable, enabling potential propagation and real-time downstream analysis using next generation sequencing (NGS) or proteomic analysis.
Highlights
Circulating tumor cells (CTCs) is a collective term to describe cancer cells of solid tumor origin found in the blood of cancer patients
Enhanced spiral microfluidic device The microfluidic device consisted of three stacked spiral microfluidic chips with two inlets and two outlets (Fig. 1B)
The multiplexed version presented here had been further enhanced to provide a device of high throughput (20 times faster) (7.5 ml in less than 5 min), high sensitivity (100% detection) (3– 1,535 CTCs) and selectivity (Mean: 750 WBCs/ml)
Summary
Circulating tumor cells (CTCs) is a collective term to describe cancer cells of solid tumor origin found in the blood of cancer patients. Current CTC capture platforms employ flow cytometry [3], fluorescence and magnetic-activated cell sorting methods [4], gradient centrifugation [5], and filtration [6,7,8]. These techniques are often limited by lengthy and complicated processing procedures, low purity and cell viability. Circulating tumor cells (CTCs) are cancer cells that can be isolated via liquid biopsy from blood and can be phenotypically and genetically characterized to provide critical information for guiding cancer treatment. Current analysis of CTCs is hindered by the throughput, selectivity and specificity of devices or assays used in CTC detection and isolation
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