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Emre Ozkumur, Ajay M Shah,
Jordan C Ciciliano,
Benjamin L Emmink,
David T Miyamoto,
Elena Brachtel,
Min Yu,
Pin-I Chen,
Bailey Morgan,
Julie Trautwein, [......],
David T Ting,
Xi Luo,
Alice T Shaw,
Aditya Bardia,
Lecia V Sequist,
David N Louis,
Shyamala Maheswaran,
Ravi Kapur,
Daniel A Haber,
Mehmet Toner
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ABSTRACT: Circulating tumor cells (CTCs) are shed into the bloodstream from primary and metastatic tumor deposits. Their isolation and analysis hold great promise for the early detection of invasive cancer and the management of advanced disease, but technological hurdles have limited their broad clinical utility. We describe an inertial focusing-enhanced microfluidic CTC capture platform, termed "CTC-iChip," that is capable of sorting rare CTCs from whole blood at 10(7) cells/s. Most importantly, the iChip is capable of isolating CTCs using strategies that are either dependent or independent of tumor membrane epitopes, and thus applicable to virtually all cancers. We specifically demonstrate the use of the iChip in an expanded set of both epithelial and nonepithelial cancers including lung, prostate, pancreas, breast, and melanoma. The sorting of CTCs as unfixed cells in solution allows for the application of high-quality clinically standardized morphological and immunohistochemical analyses, as well as RNA-based single-cell molecular characterization. The combination of an unbiased, broadly applicable, high-throughput, and automatable rare cell sorting technology with generally accepted molecular assays and cytology standards will enable the integration of CTC-based diagnostics into the clinical management of cancer.
Science translational medicine 04/2013; 5(179):179ra47. · 7.80 Impact Factor
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Min Yu,
Aditya Bardia,
Ben S Wittner,
Shannon L Stott,
Malgorzata E Smas,
David T Ting,
Steven J Isakoff,
Jordan C Ciciliano,
Marissa N Wells, Ajay M Shah,
Kyle F Concannon,
Maria C Donaldson,
Lecia V Sequist,
Elena Brachtel,
Dennis Sgroi,
Jose Baselga,
Sridhar Ramaswamy,
Mehmet Toner,
Daniel A Haber,
Shyamala Maheswaran
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ABSTRACT: Epithelial-mesenchymal transition (EMT) of adherent epithelial cells to a migratory mesenchymal state has been implicated in tumor metastasis in preclinical models. To investigate its role in human cancer, we characterized EMT in circulating tumor cells (CTCs) from breast cancer patients. Rare primary tumor cells simultaneously expressed mesenchymal and epithelial markers, but mesenchymal cells were highly enriched in CTCs. Serial CTC monitoring in 11 patients suggested an association of mesenchymal CTCs with disease progression. In an index patient, reversible shifts between these cell fates accompanied each cycle of response to therapy and disease progression. Mesenchymal CTCs occurred as both single cells and multicellular clusters, expressing known EMT regulators, including transforming growth factor (TGF)-β pathway components and the FOXC1 transcription factor. These data support a role for EMT in the blood-borne dissemination of human breast cancer.
Science 02/2013; 339(6119):580-584. · 31.20 Impact Factor
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ABSTRACT: Microfluidic systems for affinity-based cell isolation have emerged as a promising approach for the isolation of specific cells from complex matrices (i.e., circulating tumor cells in whole blood). However, these technologies remain limited by the lack of reliable methods for the innocuous recovery of surface captured cells. Here, we present a biofunctional sacrificial hydrogel coating for microfluidic chips that enables the highly efficient release of isolated cells (99% ± 1%) following gel dissolution. This covalently cross-linked alginate biopolymer system is stable in a wide variety of physiologic solutions (including EDTA treated whole blood) and may be rapidly degraded via backbone cleavage with alginate lyase. The capture and release of EpCAM expressing cancer cells using this approach was found to have no significant effect on cell viability or proliferative potential, and recovered cells were demonstrated to be compatible with downstream immunostaining and FISH analysis.
Analytical Chemistry 03/2012; 84(8):3682-8. · 5.86 Impact Factor
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Shannon L Stott,
Chia-Hsien Hsu,
Dina I Tsukrov,
Min Yu,
David T Miyamoto,
Belinda A Waltman,
S Michael Rothenberg, Ajay M Shah,
Malgorzata E Smas,
George K Korir, [......],
Daniel Winokur,
Simeon Springer,
Daniel Irimia,
Sunitha Nagrath,
Lecia V Sequist,
Richard J Lee,
Kurt J Isselbacher,
Shyamala Maheswaran,
Daniel A Haber,
Mehmet Toner
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ABSTRACT: Rare circulating tumor cells (CTCs) present in the bloodstream of patients with cancer provide a potentially accessible source for detection, characterization, and monitoring of nonhematological cancers. We previously demonstrated the effectiveness of a microfluidic device, the CTC-Chip, in capturing these epithelial cell adhesion molecule (EpCAM)-expressing cells using antibody-coated microposts. Here, we describe a high-throughput microfluidic mixing device, the herringbone-chip, or "HB-Chip," which provides an enhanced platform for CTC isolation. The HB-Chip design applies passive mixing of blood cells through the generation of microvortices to significantly increase the number of interactions between target CTCs and the antibody-coated chip surface. Efficient cell capture was validated using defined numbers of cancer cells spiked into control blood, and clinical utility was demonstrated in specimens from patients with prostate cancer. CTCs were detected in 14 of 15 (93%) patients with metastatic disease (median = 63 CTCs/mL, mean = 386 ± 238 CTCs/mL), and the tumor-specific TMPRSS2-ERG translocation was readily identified following RNA isolation and RT-PCR analysis. The use of transparent materials allowed for imaging of the captured CTCs using standard clinical histopathological stains, in addition to immunofluorescence-conjugated antibodies. In a subset of patient samples, the low shear design of the HB-Chip revealed microclusters of CTCs, previously unappreciated tumor cell aggregates that may contribute to the hematogenous dissemination of cancer.
Proceedings of the National Academy of Sciences 10/2010; 107(43):18392-7. · 9.68 Impact Factor
