A Novel Strategy for Detection and Enumeration of Circulating Rare Cell Populations in Metastatic Cancer Patients Using Automated Microfluidic Filtration and Multiplex Immunoassay.
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.
Division of Hematology-Oncology, University of California San Francisco, San Francisco, CA, 94115, United States of America.
Mark Jesus M
MJ
Magbanua
Siemens Healthcare Diagnostics, Elkhart, IN, 46516, United States of America.
Michael
M
Pugia
Division of Hematology-Oncology, University of California San Francisco, San Francisco, CA, 94115, United States of America.
Jin Sun
JS
Lee
Division of Hematology-Oncology, University of California San Francisco, San Francisco, CA, 94115, United States of America.
Marc
M
Jabon
Division of Hematology-Oncology, University of California San Francisco, San Francisco, CA, 94115, United States of America.
Victoria
V
Wang
Division of Hematology-Oncology, University of California San Francisco, San Francisco, CA, 94115, United States of America.
Matthew
M
Gubens
Siemens Healthcare Diagnostics, Elkhart, IN, 46516, United States of America.
Karen
K
Marfurt
Siemens Healthcare Diagnostics, Elkhart, IN, 46516, United States of America.
Julia
J
Pence
Siemens Healthcare Diagnostics, Elkhart, IN, 46516, United States of America.
Harwinder
H
Sidhu
Siemens Healthcare Diagnostics, Elkhart, IN, 46516, United States of America.
Arejas
A
Uzgiris
Division of Hematology-Oncology, University of California San Francisco, San Francisco, CA, 94115, United States of America.
Hope S
HS
Rugo
Division of Hematology-Oncology, University of California San Francisco, San Francisco, CA, 94115, United States of America.
John W
JW
Park
D014408:Biomarkers, Tumor / Q000235:genetics* / Q000378:metabolism; D001943:Breast Neoplasms / Q000175:diagnosis / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D016022:Case-Control Studies; D002452:Cell Count / Q000295:instrumentation / Q000379:methods; D042783:Endothelial Cells / Q000378:metabolism / Q000473:pathology; D005260:Female; D005374:Filtration; D015870:Gene Expression; D006801:Humans; D007118:Immunoassay / Q000379:methods; D008175:Lung Neoplasms / Q000175:diagnosis / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D008297:Male; D059630:Mesenchymal Stem Cells / Q000378:metabolism / Q000473:pathology; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation; D009362:Neoplasm Metastasis; D009363:Neoplasm Proteins / Q000235:genetics* / Q000378:metabolism; D009360:Neoplastic Cells, Circulating / Q000378:metabolism / Q000473:pathology*; D014411:Neoplastic Stem Cells / Q000378:metabolism / Q000473:pathology; D015203:Reproducibility of Results
D014408:Biomarkers, Tumor; D009363:Neoplasm Proteins