her2 expression and amplification in ctcs of metastatic breast cancer patients … · 2017. 12....
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HER2 Expression and Amplification in CTCsof Metastatic Breast Cancer patients
Anja Brouwer1,2,3, Pieter-Jan van Dam1,2, Jasper Van Haver2, Ellen Sluydts5, Peter Vermeulen1,5, Marc Peeters1,3, Steven Van Laere1,2, Luc Dirix2,4.
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1 Center for Oncological Research (CORE), University of Antwerp, Belgium2 Translational Cancer Research Unit (TCRU), GZA Sint Augustinus, Antwerp, Belgium3 Department of Oncology, Antwerp University Hospital (UZA), Antwerp, Belgium4 Departmetn of Oncology, GZA Sint Augustinus, Antwerp, Belgium5 HistoGeneX, Antwerp, Belgium
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Circulating Tumor Cells
• In metastatic disease multiple clones of tumorcells evolve in time and under therapy pressure.
• CTCs are known to represent metastatic disease and are an easy source for repeated tumor analysis.
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Circulating Tumor Cells
HER2 amplificationHER2 expression
HER2 expression on CTCs
HER2 amplification in CTCs
IF(CellSearch)
FISH
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Phase III trialLapatinib (TKI)
DETECT III
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Phase III trialLapatinib (TKI)
DETECT III - CTC scoring with HER2 IF
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Phase II trialT-DM1
CirCe T-DM1 - CTC scoring with HER2 FISH
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Methods
HER2 expression by IF
- CellSearch (FDA approved)
HER2 amplification by FISH
- Bioview
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Methods
HER2 expression by IF
- CellSearch (FDA approved)
HER2 amplification by FISH
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CTC
WBC
Enrichment withAnti-EpCAM
Anti-cytokeratin
Anti-CD45
HER2
Anti-HER2
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Methods
HER2 expression by IF
- CellSearch (FDA approved)
HER2 amplification by FISH
- BioView
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IF
CTC
WBC
Enrichment withAnti-EpCAM
Anti-cytokeratin
Anti-CD45
HER2
Anti-HER2
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Methods
HER2 expression by IF
- CellSearch (FDA approved)
HER2 amplification by FISH
- BioView
10
IF FISH
CTC
WBC
Enrichment withAnti-EpCAM
Anti-cytokeratin
Anti-CD45
HER2
Anti-HER2
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Cell line model
MCF-7MDA-MB-436 (0+)
BT-20 (1+)
KPL-4 (2+)
SUM190IBC-3 (3+)SKBR-3
Leukocytes
CK DAPI HER2 Bright Field
CD45
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HER2 intensity per Cell line
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1+0 2+
3+
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Automatic HER2 scoring on IF images
Patient samples:
- GZA St Augustinus: 40.000 CTCs in 128 pt samples
- Erasmus MC (NL): 10.000 CTCs in 38 pt samples
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Automatic HER2 scoring on IF images
Patient samples:
- GZA St Augustinus: 40.000 CTCs in 128 pt samples
- Erasmus MC (NL): 10.000 CTCs in 38 pt samples
Manual scoring
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HER2 scores on CTCs of 58 HER2- patient
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HER2 scores on CTCs of 24 HER2+ patient
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Automatic HER2 scoring on IF images
Patient samples:
- GZA St Augustinus: 40.000 CTCs in 128 pt samples
- Erasmus MC (NL): 10.000 CTCs in 38 pt samples
Manual scoring vs. Machine learning
17 Esteva, Nature 02/2017
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Performance of Image machine learning
ImageJPatients
Manual Scoring
0 1+ 2+ 3+
Pre
dic
tio
n
0 201 45 1 4
1+ 508 1608 83 0
2+ 7 101 170 8
3+ 2 2 13 135
Accuracy: 85,6%
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Sens Spec
0 68,8 97,1
1+ 92,3 76,0
2+ 77,4 96,9
3+ 95,2 99,7
MachineLearningPatients
Manual Scoring
0 1+ 2+ 3+
Pre
dic
tio
n
0 1239 169 3 0
1+ 562 4440 138 0
2+ 0 195 538 20
3+ 1 4 16 394
Sens Spec
0 28,0 97,7
1+ 91,6 47,8
2+ 63,7 95,6
3+ 91,8 99,4
Accuracy: 73,2%
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BioView after DAKO-IQ FISH
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SKBR-3 WBC
IF FISH
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Thanks to everyone who support this research.
Thanks to all patients who are willing to participate in our studies
CORE Clinical Trials OncologyDr. L. Dirix Dr. H. DuboisProf. S. Van Laere Anke SeytsmaProf. dr. M Peeters All research nursesDr. P. VermeulenDr. R. Salgado
Bram De LaerePieter-Jan van DamAnja BrouwerCharlotte RypensChristophe Van BerckelaerSteffi OeyenJasper Van HaverChristel VerbruggenKatrien Coen