Retracing Circulating Tumour Cells for Biomarker Characterization after Enumeration

Frandsen, Anders S.; Fabisiewicz, Anna; Jagiello-Gruszfeld, Agnieszka; Haugaard, Anastasiya S.; Petersen, Louise Munkhaus; Albrektsen, Katrine Brandt; Nejlund, Sarah; Smith, Julie; Stender, Henrik; Hillig, Thore; Söletormos, György

doi:10.5772/60995

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Retracing Circulating Tumour Cells for Biomarker Characterization after Enumeration

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Author(s): Anders S. Frandsen ¹ , Anna Fabisiewicz ² , Agnieszka Jagiello-Gruszfeld ³ , Anastasiya S. Haugaard ⁴ , Louise Munkhaus Petersen ⁴ , Katrine Brandt Albrektsen ⁴ , Sarah Nejlund ⁴ , Julie Smith ⁵ , Henrik Stender ¹ ^, ⁴ ^, ^* , Thore Hillig ⁴ ^, ⁶ , György Sölétormos ⁴ ^, ⁶

Publication date (Electronic): 30 June 2015

Journal: Journal of Circulating Biomarkers

Publisher: SAGE Publications

Keywords: CytoDisc, Ciculating tumor cells, CytoTrack, Cancer, HER2, CTC, Characterization, Enumeration, Breast cancer, Immunofluorescence, CK, Liquid biopsy, Method, Retracing, Metastasis

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Abstract

Background

Retracing and biomarker characterization of individual circulating tumour cells (CTCs) may potentially contribute to personalized metastatic cancer therapy. This is relevant when a biopsy of the metastasis is complicated or impossible to acquire.

Methods

A novel disc format was used to map and retrace individual CTCs from breast-cancer patients and nucleated cells from healthy blood donors using the CytoTrack platform. For proof of the retracing concept, CTC HER2 characterization by immunofluorescence was tested.

Results

CTCs were detected and enumerated in three of four blood samples from breast-cancer patients and the locations of each individual CTCs were mapped on the discs. Nucleated cells were retraced on seven discs with 96.6%±8.5% recovery on five fields of view on each disc. Shifting of field of view for retracing was measured to 4-29 μm. In a blood sample from a HER2-positive breast-cancer patient, CTC enumeration and mapping was followed by HER2 characterization and retracing to demonstrate downstream immunofluorescence analysis of the CTC.

Conclusion

Mapping and retracing of CTCs enables downstream analysis of individual CTCs for existing and future cancer genotypic and phenotypic biomarkers. Future studies will uncover this potential of the novel retracing technology.

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Most cited references 20

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The Hallmarks of Cancer

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Tumor cells circulate in the peripheral blood of all major carcinomas but not in healthy subjects or patients with nonmalignant diseases.

W Jeffrey Allard, Jeri Matera, M. Craig Miller … (2004)

The purpose of this study was to determine the accuracy, precision, and linearity of the CellSearch system and evaluate the number of circulating tumor cells (CTCs) per 7.5 mL of blood in healthy subjects, patients with nonmalignant diseases, and patients with a variety of metastatic carcinomas. The CellSearch system was used to enumerate CTCs in 7.5 mL of blood. Blood samples spiked with cells from tumor cell lines were used to establish analytical accuracy, reproducibility, and linearity. Prevalence of CTCs was determined in blood from 199 patients with nonmalignant diseases, 964 patients with metastatic carcinomas, and 145 healthy donors. Enumeration of spiked tumor cells was linear over the range of 5 to 1,142 cells, with an average recovery of >/=85% at each spike level. Only 1 of the 344 (0.3%) healthy and nonmalignant disease subjects had >/=2 CTCs per 7.5 mL of blood. In 2,183 blood samples from 964 metastatic carcinoma patients, CTCs ranged from 0 to 23,618 CTCs per 7.5 mL (mean, 60 +/- 693 CTCs per 7.5 mL), and 36% (781 of 2,183) of the specimens had >/=2 CTCs. Detection of >/=2 CTCs occurred at the following rates: 57% (107 of 188) of prostate cancers, 37% (489 of 1,316) of breast cancers, 37% (20 of 53) of ovarian cancers, 30% (99 of 333) of colorectal cancers, 20% (34 of 168) of lung cancers, and 26% (32 of 125) of other cancers. The CellSearch system can be standardized across multiple laboratories and may be used to determine the clinical utility of CTCs. CTCs are extremely rare in healthy subjects and patients with nonmalignant diseases but present in various metastatic carcinomas with a wide range of frequencies.

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Circulating tumor cells, disease progression, and survival in metastatic breast cancer.

Massimo Cristofanilli, G Budd, Matthew Ellis … (2004)

We tested the hypothesis that the level of circulating tumor cells can predict survival in metastatic breast cancer. In a prospective, multicenter study, we tested 177 patients with measurable metastatic breast cancer for levels of circulating tumor cells both before the patients were to start a new line of treatment and at the first follow-up visit. The progression of the disease or the response to treatment was determined with the use of standard imaging studies at the participating centers. Outcomes were assessed according to levels of circulating tumor cells at baseline, before the patients started a new treatment for metastatic disease. Patients in a training set with levels of circulating tumor cells equal to or higher than 5 per 7.5 ml of whole blood, as compared with the group with fewer than 5 circulating tumor cells per 7.5 ml, had a shorter median progression-free survival (2.7 months vs. 7.0 months, P 18 months, P 18 months; P<0.001), and the reduced proportion of patients (from 49 percent to 30 percent) in the group with an unfavorable prognosis suggested that there was a benefit from therapy. The multivariate Cox proportional-hazards regression showed that, of all the variables in the statistical model, the levels of circulating tumor cells at baseline and at the first follow-up visit were the most significant predictors of progression-free and overall survival. The number of circulating tumor cells before treatment is an independent predictor of progression-free survival and overall survival in patients with metastatic breast cancer. Copyright 2004 Massachusetts Medical Society

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Author and article information

Journal

Journal ID (nlm-ta): J Circ Biomark

Journal ID (iso-abbrev): J Circ Biomark

Journal ID (publisher-id): CBX

Journal ID (hwp): spcbx

Title: Journal of Circulating Biomarkers

Publisher: SAGE Publications (Sage UK: London, England )

ISSN (Electronic): 1849-4544

Publication date (Electronic): 30 June 2015

Publication date Collection: Jan-Dec 2015

Volume: 4

Electronic Location Identifier: 5

Affiliations

[1 ] CytoTrack ApS, Lyngby, Denmark

[2 ] Department of Translational and Molecular Oncology, The Maria Sklodowska-Curie Memorial Cancer Centre and Institute of Oncology, Warsaw, Poland

[3 ] Department of Breast Cancer and Reconstruction Surgery, The Maria Sklodowska-Curie Memorial Cancer Centre and Institute of Oncology, Warsaw, Poland

[4 ] CTC Center of Excellence, Department of Clinical Biochemistry, North Zealand Hospital, University of Copenhagen, Denmark

[5 ] Department of Technology, Faculty of Health and Technology, Metropolitan University College, Copenhagen, Denmark

[6 ] Department of Clinical Biochemistry, North Zealand Hospital, University of Copenhagen, Denmark

Author notes

[*] [* ] Corresponding author(s) E-mail: hs@ 123456cytotrack.com

Article

Publisher ID: 10.5772_60995

DOI: 10.5772/60995

PMC ID: 5572983

SO-VID: 2ebe39c7-9604-4bab-9a15-082c740c6ee8

License:

This is an open access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.