Classification of Cells in CTC-Enriched Samples by Advanced Image Analysis

Wit, Sanne de; Zeune, Leonie L.; Hiltermann, T. Jeroen N.; Groen, Harry J.M.; Dalum, Guus van; Terstappen, Leon W.M.M.

doi:10.3390/cancers10100377

Cited by 37 publications

(38 citation statements)

References 38 publications

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“…One hundred and ninety CRPC (IMMC38 clinical trial, NCT00133900), 30 450 mCRC (CAIRO II clinical trial, NCT00208546), 31,32 179 MBC (IMMC01 clinical trial), 33 and 137 NSCLC patient samples, [34][35][36] along with 93 healthy control samples (IMMC06 clinical trial, NCT00133913), 37 were included. The included samples corresponded to patients before the initiation of a new treatment.…”

Section: Patient Samplesmentioning

confidence: 99%

Tumour-derived extracellular vesicles in blood of metastatic cancer patients associate with overall survival

Nanou

Miller²,

Zeune

et al. 2020

Br J Cancer

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BACKGROUND: Circulating tumour cells (CTCs) in blood associate with overall survival (OS) of cancer patients, but they are detected in extremely low numbers. Large tumour-derived extracellular vesicles (tdEVs) in castration-resistant prostate cancer (CRPC) patients are present at around 20 times higher frequencies than CTCs and have equivalent prognostic power. In this study, we explored the presence of tdEVs in other cancers and their association with OS. METHODS:The open-source ACCEPT software was used to automatically enumerate tdEVs in digitally stored CellSearch® images obtained from previously reported CTC studies evaluating OS in 190 CRPC, 450 metastatic colorectal cancer (mCRC), 179 metastatic breast cancer (MBC) and 137 non-small cell lung cancer (NSCLC) patients before the initiation of a new treatment. RESULTS: Presence of unfavourable CTCs and tdEVs is predictive of OS, with respective hazard ratios (HRs) of 2.4 and 2.2 in CRPC, 2.7 and 2.2 in MBC, 2.3 and 1.9 in mCRC and 2.0 and 2.4 in NSCLC, respectively. CONCLUSIONS: tdEVs have equivalent prognostic value as CTCs in the investigated metastatic cancers. CRPC, mCRC, and MBC (but not NSCLC) patients with favourable CTC counts can be further prognostically stratified using tdEVs. Our data suggest that tdEVs could be used in clinical decision-making.

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Section: Patient Samplesmentioning

confidence: 99%

Tumour-derived extracellular vesicles in blood of metastatic cancer patients associate with overall survival

Nanou

Miller²,

Zeune

et al. 2020

Br J Cancer

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“…github.com/LeonieZ/ACCEPT). 27,28,32 The ACCEPT toolbox detects all events present in the images by an advanced multiscale segmentation approach and extracts several fluorescence intensities and shape measurements for every event it has found. The tdEV identified here are relatively large as they have been pelleted with the blood cell fraction after centrifugation at 800g.…”

Section: Analysis Of Tdev With Acceptmentioning

confidence: 99%

“…21 The tdEV were identified in the images from the CellSearch using the open source imaging program ACCEPT, whereas plasma from the same tube was used for cell-free ctDNA extraction followed by an untargeted tumor allele fraction analysis. [27][28][29] Moreover, for a subset of patients tumor-specific mutations were tracked in plasma DNA using deep sequencing or Safe-SeqS. 30 In total 97 NSCLC patients were included and the presence of these different biomarkers in an all-in-one liquid biopsy was explored.…”

Section: Introductionmentioning

confidence: 99%

Single tube liquid biopsy for advanced non‐small cell lung cancer

Wit

Rossi

Weber

et al. 2019

Intl Journal of Cancer

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The need for a liquid biopsy in non‐small cell lung cancer (NSCLC) patients is rapidly increasing. We studied the relation between overall survival (OS) and the presence of four cancer biomarkers from a single blood draw in advanced NSCLC patients: EpCAMhigh circulating tumor cells (CTC), EpCAMlow CTC, tumor‐derived extracellular vesicles (tdEV) and cell‐free circulating tumor DNA (ctDNA). EpCAMhigh CTC were detected with CellSearch, tdEV in the CellSearch images and EpCAMlow CTC with filtration after CellSearch. ctDNA was isolated from plasma and mutations present in the primary tumor were tracked with deep sequencing methods. In 97 patients, 21% had ≥2 EpCAMhigh CTC, 15% had ≥2 EpCAMlow CTC, 27% had ≥18 tdEV and 19% had ctDNA with ≥10% mutant allele frequency. Either one of these four biomarkers could be detected in 45% of the patients and all biomarkers were present in 2%. In 11 out of 16 patients (69%) mutations were detected in the ctDNA. Two or more unfavorable biomarkers were associated with poor OS. The presence of EpCAMhigh CTC and elevated levels of tdEV and ctDNA was associated with a poor OS; however, the presence of EpCAMlow CTC was not. This single tube approach enables simultaneous analysis of multiple biomarkers to explore their potential as a liquid biopsy.

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“…eu) open source image analysis tools (ACCEPT) are being developed and introduced to aid in the evaluation of new CTC technologies and arrive at a common CTC definition (21)(22)(23). To overcome this challenge as part of the European program CANCER-ID (www.cancer-id.…”

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confidence: 99%

“…To overcome this challenge as part of the European program CANCER-ID (www.cancer-id. eu) open source image analysis tools (ACCEPT) are being developed and introduced to aid in the evaluation of new CTC technologies and arrive at a common CTC definition (21)(22)(23). In this issue, we compared CTC scoring between operators and show that a computer-generated definition is most likely preferable to trying to have operators agree on a definition (24).…”

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confidence: 99%

CTC Technologies and Tools

Coumans

Dalum

Terstappen³

2018

Cytometry Pt A

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CANCER cells can enter the blood stream. Some of these circulating tumor cells (CTC) extravasate and form distant metastases which ultimately lead to the death of the patient. CTC have been observed quite some time ago (1-3). The major difficulty for the identification of CTC is their extremely low frequency (~1/ml in metastatic patients). When no CTC were detected in a tube of blood of a patient with metastases one always wonders whether they were not present or whether they were missed by the detection technology. Another hurdle is the extensive heterogeneity of the physical and immunological appearance of CTC within and between patients.Tumor cell lines play a pivotal role in both informing technology developers of the probable physical and immunological properties of CTC, as well as in the determination of the analytical accuracy, reproducibility, and linearity of any developed technology. However, tumor cell lines are not CTC. Excellent performance on tumor cell lines does not warrant that (1) the developed technology will actually identify CTC in blood samples of cancer patients nor that (2) these CTC relate to clinical outcome nor that (3) information relevant for treatment can be extracted.At the start of the trajectory toward the FDA cleared CellSearch system very little was known about the CTC frequency and their physical and immunological appearance. CTC enrichment in 10 ml of blood was performed by manual immunomagnetic enrichment targeting EpCAM using the GA73.3 antibody. The enriched cell suspension was fluorescently labeled with the PE-labeled CAM5.2 antibody recognizing cytokeratins 7 and 8, a PerCP-labeled antibody recognizing CD45 and a nucleic acid dye that could be measured in the FITC channel of a flowcytometer. Using this approach, it was first shown that CTC indeed could be detected in 10 ml blood of cancer patients with metastatic disease and at a lesser frequency in patients with no detectable metastasis while few "CTC" were detected in healthy controls (4-8). These results were sufficiently encouraging to further develop the CTC technology and design and conduct clinical studies to explore the clinical utility of CTC. The commercial CellSearch system that was ultimately developed included (1) a blood collection tube to preserve the fragile CTC for a period of up to 96 h, (2) an automated sample preparation system in which (3) the EpCAM antibody was replaced by VU1D9, (4) CAM5.2 cytokeratin antibody was replaced by C11 and A.53B/A2 to increase the range of cytokeratins, (5) the CD45-PerCP was switched to CD45-APC, (6) the nucleic acid dye was switched to DAPI, and (7) the flow cytometer replaced by a semi-automated fluorescence microscope with dedicated software to present CTC candidates to a reviewer. Thanks to an excellent team of reagent developers and engineers the system was reliable and provided accurate results over a large range of spiked tumor cells in blood with a high sensitivity and specificity. Whereas few "CTC" were detected in healthy donors and patients with benign disease...

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Classification of Cells in CTC-Enriched Samples by Advanced Image Analysis

Cited by 37 publications

References 38 publications

Tumour-derived extracellular vesicles in blood of metastatic cancer patients associate with overall survival

Tumour-derived extracellular vesicles in blood of metastatic cancer patients associate with overall survival

Single tube liquid biopsy for advanced non‐small cell lung cancer

CTC Technologies and Tools

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