Quantifying HER-2 expression on Circulating Tumor Cells by ACCEPT

Leonie Laura Zeune, Guus van Dalum, C. Decraene, C. Proudhon, T. Fehm, Hans Neubauer, B. Rack, Marianna Alunni-fabbroni, L.W.M.M. Terstappen, Stephanus A. van Gils, Christoph Brune

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Abstract

Circulating tumor cells (CTCs) isolated from blood can be probed for the expression of treatment targets. Immunofluorescence is often used for both the enumeration of CTC and the determination of protein expression levels related to treatment targets. Accurate and reproducible assessment of such treatment target expression levels is essential for their use in the clinic. To enable this, an open source image analysis program named ACCEPT was developed in the EU-FP7 CTCTrap and CANCER-ID programs. Here its application is shown on a retrospective cohort of 132 metastatic breast cancer patients from which blood samples were processed by CellSearch® and stained for HER-2 expression as additional marker. Images were digitally stored and reviewers identified a total of 4084 CTCs. CTC’s HER-2 expression was determined in the thumbnail images by ACCEPT. 150 of these images were selected and sent to six independent investigators to score the HER-2 expression with and without ACCEPT. Concordance rate of the operators’ scoring results for HER-2 on CTCs was 30% and could be increased using the ACCEPT tool to 51%. Automated assessment of HER-2 expression by ACCEPT on 4084 CTCs of 132 patients showed 8 (6.1%) patients with all CTCs expressing HER-2, 14 (10.6%) patients with no CTC expressing HER-2 and 110 (83.3%) patients with CTCs showing a varying HER-2 expression level. In total 1576 CTCs were determined HER-2 positive. We conclude that the use of image analysis enables a more reproducible quantification of treatment targets on CTCs and leads the way to fully automated and reproducible approaches.
Original languageEnglish
Article numbere0186562
Number of pages12
JournalPLoS ONE
Volume12
Issue number10
DOIs
Publication statusPublished - 30 Oct 2017

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Circulating Neoplastic Cells
Tumors
Cells
Image analysis
neoplasm cells
Blood
image analysis
operator regions
blood
Therapeutics
breast neoplasms
fluorescent antibody technique
Fluorescent Antibody Technique
protein synthesis
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Zeune, L. L., van Dalum, G., Decraene, C., Proudhon, C., Fehm, T., Neubauer, H., ... Brune, C. (2017). Quantifying HER-2 expression on Circulating Tumor Cells by ACCEPT. PLoS ONE, 12(10), [e0186562]. https://doi.org/10.1371/journal.pone.0186562
Zeune, Leonie Laura ; van Dalum, Guus ; Decraene, C. ; Proudhon, C. ; Fehm, T. ; Neubauer, Hans ; Rack, B. ; Alunni-fabbroni, Marianna ; Terstappen, L.W.M.M. ; van Gils, Stephanus A. ; Brune, Christoph . / Quantifying HER-2 expression on Circulating Tumor Cells by ACCEPT. In: PLoS ONE. 2017 ; Vol. 12, No. 10.
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abstract = "Circulating tumor cells (CTCs) isolated from blood can be probed for the expression of treatment targets. Immunofluorescence is often used for both the enumeration of CTC and the determination of protein expression levels related to treatment targets. Accurate and reproducible assessment of such treatment target expression levels is essential for their use in the clinic. To enable this, an open source image analysis program named ACCEPT was developed in the EU-FP7 CTCTrap and CANCER-ID programs. Here its application is shown on a retrospective cohort of 132 metastatic breast cancer patients from which blood samples were processed by CellSearch{\circledR} and stained for HER-2 expression as additional marker. Images were digitally stored and reviewers identified a total of 4084 CTCs. CTC’s HER-2 expression was determined in the thumbnail images by ACCEPT. 150 of these images were selected and sent to six independent investigators to score the HER-2 expression with and without ACCEPT. Concordance rate of the operators’ scoring results for HER-2 on CTCs was 30{\%} and could be increased using the ACCEPT tool to 51{\%}. Automated assessment of HER-2 expression by ACCEPT on 4084 CTCs of 132 patients showed 8 (6.1{\%}) patients with all CTCs expressing HER-2, 14 (10.6{\%}) patients with no CTC expressing HER-2 and 110 (83.3{\%}) patients with CTCs showing a varying HER-2 expression level. In total 1576 CTCs were determined HER-2 positive. We conclude that the use of image analysis enables a more reproducible quantification of treatment targets on CTCs and leads the way to fully automated and reproducible approaches.",
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Zeune, LL, van Dalum, G, Decraene, C, Proudhon, C, Fehm, T, Neubauer, H, Rack, B, Alunni-fabbroni, M, Terstappen, LWMM, van Gils, SA & Brune, C 2017, 'Quantifying HER-2 expression on Circulating Tumor Cells by ACCEPT' PLoS ONE, vol. 12, no. 10, e0186562. https://doi.org/10.1371/journal.pone.0186562

Quantifying HER-2 expression on Circulating Tumor Cells by ACCEPT. / Zeune, Leonie Laura; van Dalum, Guus ; Decraene, C.; Proudhon, C.; Fehm, T.; Neubauer, Hans; Rack, B.; Alunni-fabbroni, Marianna; Terstappen, L.W.M.M.; van Gils, Stephanus A.; Brune, Christoph .

In: PLoS ONE, Vol. 12, No. 10, e0186562, 30.10.2017.

Research output: Contribution to journalArticleAcademicpeer-review

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AU - Zeune, Leonie Laura

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AU - Proudhon, C.

AU - Fehm, T.

AU - Neubauer, Hans

AU - Rack, B.

AU - Alunni-fabbroni, Marianna

AU - Terstappen, L.W.M.M.

AU - van Gils, Stephanus A.

AU - Brune, Christoph

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AB - Circulating tumor cells (CTCs) isolated from blood can be probed for the expression of treatment targets. Immunofluorescence is often used for both the enumeration of CTC and the determination of protein expression levels related to treatment targets. Accurate and reproducible assessment of such treatment target expression levels is essential for their use in the clinic. To enable this, an open source image analysis program named ACCEPT was developed in the EU-FP7 CTCTrap and CANCER-ID programs. Here its application is shown on a retrospective cohort of 132 metastatic breast cancer patients from which blood samples were processed by CellSearch® and stained for HER-2 expression as additional marker. Images were digitally stored and reviewers identified a total of 4084 CTCs. CTC’s HER-2 expression was determined in the thumbnail images by ACCEPT. 150 of these images were selected and sent to six independent investigators to score the HER-2 expression with and without ACCEPT. Concordance rate of the operators’ scoring results for HER-2 on CTCs was 30% and could be increased using the ACCEPT tool to 51%. Automated assessment of HER-2 expression by ACCEPT on 4084 CTCs of 132 patients showed 8 (6.1%) patients with all CTCs expressing HER-2, 14 (10.6%) patients with no CTC expressing HER-2 and 110 (83.3%) patients with CTCs showing a varying HER-2 expression level. In total 1576 CTCs were determined HER-2 positive. We conclude that the use of image analysis enables a more reproducible quantification of treatment targets on CTCs and leads the way to fully automated and reproducible approaches.

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Zeune LL, van Dalum G, Decraene C, Proudhon C, Fehm T, Neubauer H et al. Quantifying HER-2 expression on Circulating Tumor Cells by ACCEPT. PLoS ONE. 2017 Oct 30;12(10). e0186562. https://doi.org/10.1371/journal.pone.0186562