A Novel Strategy for Detection and Enumeration of Circulating Rare Cell Populations in Metastatic Cancer Patients Using Automated Microfluidic Filtration and Multiplex Immunoassay

• Published in: PloS one Vol. 10; no. 10; p. e0141166
• Main Authors: Magbanua, Mark Jesus M, Pugia, Michael, Lee, Jin Sun, Jabon, Marc, Wang, Victoria, Gubens, Matthew, Marfurt, Karen, Pence, Julia, Sidhu, Harwinder, Uzgiris, Arejas, Rugo, Hope S, Park, John W
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 23.10.2015; Public Library of Science (PLoS)
• Subjects: Antigens; Automatic control; Biomarkers, Tumor - genetics; Biomarkers, Tumor - metabolism; Blood; Blood circulation; Breast cancer; Breast Neoplasms - diagnosis; Breast Neoplasms - genetics; Breast Neoplasms - metabolism; Breast Neoplasms - pathology; Cancer; Case-Control Studies; Cell Count - instrumentation; Cell Count - methods; Endothelial cells; Endothelial Cells - metabolism; Endothelial Cells - pathology; Enumeration; Feasibility studies; Female; Filtration; Gene Expression; Gynecology; Hematology; Humans; Immunoassay; Immunoassay - methods; Life assessment; Lung cancer; Lung diseases; Lung Neoplasms - diagnosis; Lung Neoplasms - genetics; Lung Neoplasms - metabolism; Lung Neoplasms - pathology; Male; Medical research; Medical treatment; Mesenchymal Stromal Cells - metabolism; Mesenchymal Stromal Cells - pathology; Mesenchyme; Metastases; Metastasis; Microfluidic Analytical Techniques - instrumentation; Microfluidics; Multiplexing; Neoplasm Metastasis; Neoplasm Proteins - genetics; Neoplasm Proteins - metabolism; Neoplastic Cells, Circulating - metabolism; Neoplastic Cells, Circulating - pathology; Neoplastic Stem Cells - metabolism; Neoplastic Stem Cells - pathology; Oncology; Patients; Populations; Pore size; Reproducibility; Reproducibility of Results; Stem cells; Tumor cells; United States > US; San Francisco California; California
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0141166

Size selection via filtration offers an antigen-independent approach for the enrichment of rare cell populations in blood of cancer patients. We evaluated the performance of a novel approach for multiplex rare cell detection in blood samples from metastatic breast (n = 19) and lung cancer patients (n = 21), and healthy controls (n = 30) using an automated microfluidic filtration and multiplex immunoassay strategy. Captured cells were enumerated after sequential staining for specific markers to identify circulating tumor cells (CTCs), circulating mesenchymal cells (CMCs), putative circulating stem cells (CSCs), and circulating endothelial cells (CECs). Preclinical validation experiments using cancer cells spiked into healthy blood demonstrated high recovery rate (mean = 85%) and reproducibility of the assay. In clinical studies, CTCs and CMCs were detected in 35% and 58% of cancer patients, respectively, and were largely absent from healthy controls (3%, p = 0.001). Mean levels of CTCs were significantly higher in breast than in lung cancer patients (p = 0.03). Fifty-three percent (53%) of cancer patients harbored putative CSCs, while none were detectable in healthy controls (p<0.0001). In contrast, CECs were observed in both cancer and control groups. Direct comparison of CellSearch® vs. our microfluidic filter method revealed moderate correlation (R2 = 0.46, kappa = 0.47). Serial blood analysis in breast cancer patients demonstrated the feasibility of monitoring circulating rare cell populations over time. Simultaneous assessment of CTCs, CMCs, CSCs and CECs may provide new tools to study mechanisms of disease progression and treatment response/resistance.