EMT-independent Detection of Circulating Tumor Cells in Human Blood Samples and Pre-clinical Mouse Models of Metastasis

Overview

Journal Clin Exp Metastasis

Specialty Oncology

Date 2021 Jan 8

PMID 33415568

Citations 13

Authors

Jenna Kitz

David Goodale

Carl Postenka

Lori E Lowes

Alison L Allan

Affiliations

Soon will be listed here.

Abstract

Circulating tumor cells (CTCs) present an opportunity to detect/monitor metastasis throughout disease progression. The CellSearch® is currently the only FDA-approved technology for CTC detection in patients. The main limitation of this system is its reliance on epithelial markers for CTC isolation/enumeration, which reduces its ability to detect more aggressive mesenchymal CTCs that are generated during metastasis via epithelial-to-mesenchymal transition (EMT). This Technical Note describes and validates two EMT-independent CTC analysis protocols; one for human samples using Parsortix® and one for mouse samples using VyCap. Parsortix® identifies significantly more mesenchymal human CTCs compared to the clinical CellSearch® test, and VyCap identifies significantly more CTCs compared to our mouse CellSearch® protocol regardless of EMT status. Recovery and downstream molecular characterization of CTCs is highly feasible using both Parsortix® and VyCap. The described CTC protocols can be used by investigators to study CTC generation, EMT and metastasis in both pre-clinical models and clinical samples.

Citing Articles

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References

Coumans F, van Dalum G, Beck M, Terstappen L . Filter characteristics influencing circulating tumor cell enrichment from whole blood. PLoS One. 2013; 8(4):e61770. PMC: 3634026. DOI: 10.1371/journal.pone.0061770. View

Mittal V . Epithelial Mesenchymal Transition in Tumor Metastasis. Annu Rev Pathol. 2018; 13:395-412. DOI: 10.1146/annurev-pathol-020117-043854. View

Zweitzig D, Tibbe A, Nguyen A, van Rijn C, Kopnitsky M, Cichonski K . Feasibility of a simple microsieve-based immunoassay platform. J Immunol Methods. 2016; 437:21-7. DOI: 10.1016/j.jim.2016.07.002. View

Price J, Polyzos A, Zhang R, Daniels L . Tumorigenicity and metastasis of human breast carcinoma cell lines in nude mice. Cancer Res. 1990; 50(3):717-21. View

KAIGHN M, Narayan K, Ohnuki Y, Lechner J, Jones L . Establishment and characterization of a human prostatic carcinoma cell line (PC-3). Invest Urol. 1979; 17(1):16-23. View

Xu L, Mao X, Imrali A, Syed F, Mutsvangwa K, Berney D . Optimization and Evaluation of a Novel Size Based Circulating Tumor Cell Isolation System. PLoS One. 2015; 10(9):e0138032. PMC: 4580600. DOI: 10.1371/journal.pone.0138032. View

Gorges T, Tinhofer I, Drosch M, Rose L, Zollner T, Krahn T . Circulating tumour cells escape from EpCAM-based detection due to epithelial-to-mesenchymal transition. BMC Cancer. 2012; 12:178. PMC: 3502112. DOI: 10.1186/1471-2407-12-178. View

Neumann M, Bender S, Krahn T, Schlange T . ctDNA and CTCs in Liquid Biopsy - Current Status and Where We Need to Progress. Comput Struct Biotechnol J. 2018; 16:190-195. PMC: 6024152. DOI: 10.1016/j.csbj.2018.05.002. View

Albanese C, Rodriguez O, VanMeter J, Fricke S, Rood B, Lee Y . Preclinical magnetic resonance imaging and systems biology in cancer research: current applications and challenges. Am J Pathol. 2012; 182(2):312-8. PMC: 3969503. DOI: 10.1016/j.ajpath.2012.09.024. View

10.

Chambers A, Groom A, MacDonald I . Dissemination and growth of cancer cells in metastatic sites. Nat Rev Cancer. 2002; 2(8):563-72. DOI: 10.1038/nrc865. View

11.

Hvichia G, Parveen Z, Wagner C, Janning M, Quidde J, Stein A . A novel microfluidic platform for size and deformability based separation and the subsequent molecular characterization of viable circulating tumor cells. Int J Cancer. 2016; 138(12):2894-904. PMC: 5069649. DOI: 10.1002/ijc.30007. View

12.

Bulfoni M, Gerratana L, Del Ben F, Marzinotto S, Sorrentino M, Turetta M . In patients with metastatic breast cancer the identification of circulating tumor cells in epithelial-to-mesenchymal transition is associated with a poor prognosis. Breast Cancer Res. 2016; 18(1):30. PMC: 4784394. DOI: 10.1186/s13058-016-0687-3. View

13.

Cristofanilli M, Budd G, Ellis M, Stopeck A, Matera J, Miller M . Circulating tumor cells, disease progression, and survival in metastatic breast cancer. N Engl J Med. 2004; 351(8):781-91. DOI: 10.1056/NEJMoa040766. View

14.

de Bono J, Scher H, Montgomery R, Parker C, Miller M, Tissing H . Circulating tumor cells predict survival benefit from treatment in metastatic castration-resistant prostate cancer. Clin Cancer Res. 2008; 14(19):6302-9. DOI: 10.1158/1078-0432.CCR-08-0872. View

15.

Cohen S, Punt C, Iannotti N, Saidman B, Sabbath K, Gabrail N . Prognostic significance of circulating tumor cells in patients with metastatic colorectal cancer. Ann Oncol. 2009; 20(7):1223-9. DOI: 10.1093/annonc/mdn786. View

16.

Pailler E, Oulhen M, Billiot F, Galland A, Auger N, Faugeroux V . Method for semi-automated microscopy of filtration-enriched circulating tumor cells. BMC Cancer. 2016; 16:477. PMC: 4946105. DOI: 10.1186/s12885-016-2461-4. View

17.

Stevens M, Oomens L, Broekmaat J, Weersink J, Abali F, Swennenhuis J . VyCAP's puncher technology for single cell identification, isolation, and analysis. Cytometry A. 2018; 93(12):1255-1259. DOI: 10.1002/cyto.a.23631. View

18.

Seyfried T, Huysentruyt L . On the origin of cancer metastasis. Crit Rev Oncog. 2012; 18(1-2):43-73. PMC: 3597235. DOI: 10.1615/critrevoncog.v18.i1-2.40. View

19.

Lindsay C, Le Moulec S, Billiot F, Loriot Y, Ngo-Camus M, Vielh P . Vimentin and Ki67 expression in circulating tumour cells derived from castrate-resistant prostate cancer. BMC Cancer. 2016; 16:168. PMC: 4770547. DOI: 10.1186/s12885-016-2192-6. View

20.

Jolly M, Boareto M, Huang B, Jia D, Lu M, Ben-Jacob E . Implications of the Hybrid Epithelial/Mesenchymal Phenotype in Metastasis. Front Oncol. 2015; 5:155. PMC: 4507461. DOI: 10.3389/fonc.2015.00155. View