A Microfluidic Device for Label-free, Physical Capture of Circulating Tumor Cell Clusters

Overview

Journal Nat Methods

Specialties Biomedical Engineering
Pathology

Date 2015 May 19

PMID 25984697

Citations 289

Authors

A Fatih Sarioglu

Nicola Aceto

Nikola Kojic

Maria C Donaldson

Mahnaz Zeinali

Bashar Hamza

Amanda Engstrom

Huili Zhu

Tilak K Sundaresan

David T Miyamoto

Xi Luo

Aditya Bardia

Ben S Wittner

Sridhar Ramaswamy

Toshi Shioda

David T Ting

Shannon L Stott

Ravi Kapur

Shyamala Maheswaran

Daniel A Haber

Mehmet Toner

Affiliations

Soon will be listed here.

Abstract

Cancer cells metastasize through the bloodstream either as single migratory circulating tumor cells (CTCs) or as multicellular groupings (CTC clusters). Existing technologies for CTC enrichment are designed to isolate single CTCs, and although CTC clusters are detectable in some cases, their true prevalence and significance remain to be determined. Here we developed a microchip technology (the Cluster-Chip) to capture CTC clusters independently of tumor-specific markers from unprocessed blood. CTC clusters are isolated through specialized bifurcating traps under low-shear stress conditions that preserve their integrity, and even two-cell clusters are captured efficiently. Using the Cluster-Chip, we identified CTC clusters in 30-40% of patients with metastatic breast or prostate cancer or with melanoma. RNA sequencing of CTC clusters confirmed their tumor origin and identified tissue-derived macrophages within the clusters. Efficient capture of CTC clusters will enable the detailed characterization of their biological properties and role in metastasis.

Citing Articles

Label-Free and Rapid Microfluidic Design Rules for Circulating Tumor Cell Enrichment and Isolation: A Review and Simulation Analysis.

Mansor M, Yang C, Chong K, Jamrus M, Liu K, Yu M ACS Omega. 2025; 10(7):6306-6322.

PMID: 40028152 PMC: 11866005. DOI: 10.1021/acsomega.4c08606.

Microfluidic technologies for enhancing the potency, predictability and affordability of adoptive cell therapies.

Wang Z, Kelley S Nat Biomed Eng. 2025; .

PMID: 39953325 DOI: 10.1038/s41551-024-01315-2.

Modeling the dynamics of circulating tumor cell clusters inside a microfluidic channel.

Ezeobidi E, Truszkowska A Biomicrofluidics. 2025; 19(1):014103.

PMID: 39949346 PMC: 11821273. DOI: 10.1063/5.0249165.

Gravity-based microfiltration reveals unexpected prevalence of circulating tumor cell clusters in ovarian and colorectal cancer.

Meunier A, Hernandez-Castro J, Chahley N, Communal L, Kheireddine S, Koushki N Commun Med (Lond). 2025; 5(1):33.

PMID: 39900650 PMC: 11790846. DOI: 10.1038/s43856-024-00702-9.

Circulating tumour cell clusters: isolation, biological significance and therapeutic implications.

Yang Y, Huang G, Lian J, Long C, Zhao B, Liu X BMJ Oncol. 2025; 3(1):e000437.

PMID: 39886139 PMC: 11557725. DOI: 10.1136/bmjonc-2024-000437.

References

Miyamoto D, Lee R, Stott S, Ting D, Wittner B, Ulman M . Androgen receptor signaling in circulating tumor cells as a marker of hormonally responsive prostate cancer. Cancer Discov. 2012; 2(11):995-1003. PMC: 3508523. DOI: 10.1158/2159-8290.CD-12-0222. View

Yu M, Ting D, Stott S, Wittner B, Ozsolak F, Paul S . RNA sequencing of pancreatic circulating tumour cells implicates WNT signalling in metastasis. Nature. 2012; 487(7408):510-3. PMC: 3408856. DOI: 10.1038/nature11217. View

Hou H, Warkiani M, Khoo B, Li Z, Soo R, Tan D . Isolation and retrieval of circulating tumor cells using centrifugal forces. Sci Rep. 2013; 3:1259. PMC: 3569917. DOI: 10.1038/srep01259. View

Ozkumur E, Shah A, Ciciliano J, Emmink B, Miyamoto D, Brachtel E . Inertial focusing for tumor antigen-dependent and -independent sorting of rare circulating tumor cells. Sci Transl Med. 2013; 5(179):179ra47. PMC: 3760275. DOI: 10.1126/scitranslmed.3005616. View

Coumans F, van Dalum G, Beck M, Terstappen L . Filtration parameters influencing circulating tumor cell enrichment from whole blood. PLoS One. 2013; 8(4):e61774. PMC: 3637225. DOI: 10.1371/journal.pone.0061774. View

Inwald E, Klinkhammer-Schalke M, Hofstadter F, Zeman F, Koller M, Gerstenhauer M . Ki-67 is a prognostic parameter in breast cancer patients: results of a large population-based cohort of a cancer registry. Breast Cancer Res Treat. 2013; 139(2):539-52. PMC: 3669503. DOI: 10.1007/s10549-013-2560-8. View

Vona G, Sabile A, Louha M, Sitruk V, Romana S, Schutze K . Isolation by size of epithelial tumor cells : a new method for the immunomorphological and molecular characterization of circulatingtumor cells. Am J Pathol. 2000; 156(1):57-63. PMC: 1868645. DOI: 10.1016/S0002-9440(10)64706-2. View

Hekmat O, Munk S, Fogh L, Yadav R, Francavilla C, Horn H . TIMP-1 increases expression and phosphorylation of proteins associated with drug resistance in breast cancer cells. J Proteome Res. 2013; 12(9):4136-51. DOI: 10.1021/pr400457u. View

Yoon H, Kim T, Zhang Z, Azizi E, Pham T, Paoletti C . Sensitive capture of circulating tumour cells by functionalized graphene oxide nanosheets. Nat Nanotechnol. 2013; 8(10):735-41. PMC: 4017624. DOI: 10.1038/nnano.2013.194. View

10.

Luo X, Mitra D, Sullivan R, Wittner B, Kimura A, Pan S . Isolation and molecular characterization of circulating melanoma cells. Cell Rep. 2014; 7(3):645-53. PMC: 4079008. DOI: 10.1016/j.celrep.2014.03.039. View

11.

Kattlove H, Alexander B . The effect of cold on platelets. I. Cold-induced platelet aggregation. Blood. 1971; 38(1):39-48. View

12.

Molnar B, Ladanyi A, Tanko L, Sreter L, Tulassay Z . Circulating tumor cell clusters in the peripheral blood of colorectal cancer patients. Clin Cancer Res. 2001; 7(12):4080-5. View

13.

Aceto N, Bardia A, Miyamoto D, Donaldson M, Wittner B, Spencer J . Circulating tumor cell clusters are oligoclonal precursors of breast cancer metastasis. Cell. 2014; 158(5):1110-1122. PMC: 4149753. DOI: 10.1016/j.cell.2014.07.013. View

14.

Fidler I . The relationship of embolic homogeneity, number, size and viability to the incidence of experimental metastasis. Eur J Cancer (1965). 1973; 9(3):223-7. DOI: 10.1016/s0014-2964(73)80022-2. View

15.

Brandt B, Junker R, Griwatz C, Heidl S, Brinkmann O, Semjonow A . Isolation of prostate-derived single cells and cell clusters from human peripheral blood. Cancer Res. 1996; 56(20):4556-61. View

16.

Watanabe S . The metastasizability of tumor cells. Cancer. 1954; 7(2):215-23. DOI: 10.1002/1097-0142(195403)7:2<215::aid-cncr2820070203>3.0.co;2-6. View

17.

Wurtz S, Schrohl A, Sorensen N, Lademann U, Christensen I, Mouridsen H . Tissue inhibitor of metalloproteinases-1 in breast cancer. Endocr Relat Cancer. 2005; 12(2):215-27. DOI: 10.1677/erc.1.00719. View

18.

Wyckoff J, Wang Y, Lin E, Li J, Goswami S, Stanley E . Direct visualization of macrophage-assisted tumor cell intravasation in mammary tumors. Cancer Res. 2007; 67(6):2649-56. DOI: 10.1158/0008-5472.CAN-06-1823. View

19.

Nagrath S, Sequist L, Maheswaran S, Bell D, Irimia D, Ulkus L . Isolation of rare circulating tumour cells in cancer patients by microchip technology. Nature. 2007; 450(7173):1235-9. PMC: 3090667. DOI: 10.1038/nature06385. View

20.

Pantel K, Brakenhoff R, Brandt B . Detection, clinical relevance and specific biological properties of disseminating tumour cells. Nat Rev Cancer. 2008; 8(5):329-40. DOI: 10.1038/nrc2375. View