Abstract
Automated methods are needed to facilitate high-throughput and reproducible scoring of Ki67 and othermarkers in breast cancer tissue microarrays (TMAs) in large-scale studies. To address this need, we developedan automated protocol for Ki67 scoring and evaluated its performance in studies from the Breast CancerAssociation Consortium. We utilized 166 TMAs containing 16,953 tumour cores representing 9,059 breastcancer cases, from 13 studies, with information on other clinical and pathological characteristics. TMAs werestained for Ki67 using standard immunohistochemical procedures, and scanned and digitized using the Ariolsystem. An automated algorithm was developed for the scoring of Ki67, and scores were compared to com-puter assisted visual (CAV) scores in a subset of 15 TMAs in a training set. We also assessed the correlationbetween automated Ki67 scores and other clinical and pathological characteristics. Overall, we observed gooddiscriminatory accuracy (AUC585%) and good agreement (kappa50.64) between the automated and CAVscoring methods in the training set. The performance of the automated method varied by TMA (kappa range50.37–0.87) and study (kappa range50.39–0.69). The automated method performed better in satisfactorycores (kappa50.68) than suboptimal (kappa50.51) cores (p-value for comparison50.005); and amongcores with higher total nuclei counted by the machine (4,000–4,500 cells: kappa50.78) than those withlower counts (50–500 cells: kappa50.41;p-value50.010). Among the 9,059 cases in this study, the corre-lations between automated Ki67 and clinical and pathological characteristics were found to be in the expecteddirections. Our findings indicate that automated scoring of Ki67 can be an efficient method to obtain goodquality data across large numbers of TMAs from multicentre studies. However, robust algorithm developmentand rigorous pre- and post-analytical quality control procedures are necessary in order to ensure satisfactoryperformance.
| Original language | English |
|---|---|
| Pages (from-to) | 138-153 |
| Number of pages | 16 |
| Journal | The Journal of Pathology: Clinical Research |
| Volume | 2 |
| Issue number | 3 |
| DOIs | |
| Publication status | Published - Jul 2016 |
| Externally published | Yes |
Bibliographical note
Funding Information:The MARIE study was supported by the Deutsche Krebshilfe e.V. [70-2892-BR I, 106332, 108253, 108419], the Hamburg Cancer Society, the German Cancer Research Center (DKFZ) and the Federal Ministry of Education and Research (BMBF) Germany [01KH0402].
Funding Information:
SEARCH is funded by programme grant from Cancer Research UK [C490/A10124. C490/A16561] and supported by the UK National Institute for Health Research Biomedical Research Centre at the University of Cambridge. Part of this work was supported by the European Community’s Seventh Framework Programme under grant agreement number 223175 (grant number HEALTH-F2-2009223175) (COGS).
Funding Information:
PBCS was funded by Intramural Research Funds of the National Cancer Institute, Department of Health and Human Services, USA.
Funding Information:
The RBCS was funded by the Dutch Cancer Society (DDHK 2004-3124, DDHK 2009-4318).
Funding Information:
ABCS was supported by the Dutch Cancer Society [grants NKI 2007-3839; 2009-4363]; BBMRI-NL, which is a Research Infrastructure financed by the Dutch government (NWO 184.021.007); and the Dutch National Genomics Initiative.
Funding Information:
We wish to thank Heather Thorne, Eveline Nieder-mayr, all the kConFab research nurses and staff, the heads and staff of the Family Cancer Clinics, and the Clinical Follow Up Study (which has received funding from the NHMRC, the National Breast Cancer Foundation, Cancer Australia, and the National Institute of Health (USA)) for their contributions to this resource, and the many families who contribute to kConFab. kCon-Fab is supported by a grant from the National Breast Cancer Foundation, and previously by the National Health and Medical Research Council (NHMRC), the Queensland Cancer Fund, the Cancer Councils of New South Wales, Victoria, Tasmania and South Australia, and the Cancer Foundation of Western Australia.
Funding Information:
CNIO-BCS was supported by the Genome Spain Foundation, the Red Temática de Investigación Coop-erativa en Cáncer and grants from the Asociación Espa-ola Contra el Cáncer and the Fondo de Investigación Sanitario (PI11/00923 and PI081120). The Human Genotyping-CEGEN Unit (CNIO) is supported by the Instituto de Salud Carlos III.
Funding Information:
The MCBCS was supported by an NIH Specialized Program of Research Excellence (SPORE) in Breast Cancer [CA116201], the Breast Cancer Research Foundation, the Mayo Clinic Breast Cancer Registry and a generous gift from the David F. and Margaret T. Grohne Family Foundation and the Ting Tsung and Wei Fong Chao Foundation.
Funding Information:
The ESTHER study was supported by a grant from the Baden Wurttemberg Ministry of Science, Research and Arts. Additional cases were recruited in the context of the VERDI study, which was supported by a grant from the German Cancer Aid (Deutsche Krebshilfe).
Funding Information:
The KBCP was financially supported by the special Government Funding (EVO) of Kuopio University Hospital grants, Cancer Fund of North Savo, the Finnish Cancer Organizations, the Academy of Finland and by the strategic funding of the University of Eastern Finland.
Funding Information:
The UKBGS is funded by Breakthrough Breast Cancer and the Institute of Cancer Research (ICR), London. ICR acknowledges NHS funding to the NIHR Biomedical Research Centre.
Funding Information:
ORIGO authors thank E. Krol-Warmerdam, and J. Blom; The contributing studies were funded by grants from the Dutch Cancer Society (UL1997-1505) and the Biobanking and Biomolecular Resources Research Infrastructure (BBMRI-NL CP16).
Funding Information:
ABCS was supported by the Dutch Cancer Society [grants NKI 2007-3839; 2009-4363]; BBMRI-NL, which is a Research Infrastructure financed by the Dutch government (NWO 184.021.007); and the Dutch National Genomics Initiative. CNIO-BCS was supported by the Genome Spain Foundation, the Red Temática de Investigación Cooperativa en Cáncer and grants from the Asociación Española Contra el Cáncer and the Fondo de Investigación Sanitario (PI11/00923 and PI081120). The Human Genotyping-CEGEN Unit (CNIO) is supported by the Instituto de Salud Carlos III. The ESTHER study was supported by a grant from the Baden Württemberg Ministry of Science, Research and Arts. Additional cases were recruited in the context of the VERDI study, which was supported by a grant from the German Cancer Aid (Deutsche Krebshilfe). The KBCP was financially supported by the special Government Funding (EVO) of Kuopio University Hospital grants, Cancer Fund of North Savo, the Finnish Cancer Organizations, the Academy of Finland and by the strategic funding of the University of Eastern Finland. We wish to thank Heather Thorne, Eveline Niedermayr, all the kConFab research nurses and staff, the heads and staff of the Family Cancer Clinics, and the Clinical Follow Up Study (which has received funding from the NHMRC, the National Breast Cancer Foundation, Cancer Australia, and the National Institute of Health (USA)) for their contributions to this resource, and the many families who contribute to kConFab. kConFab is supported by a grant from the National Breast Cancer Foundation, and previously by the National Health and Medical Research Council (NHMRC), the Queensland Cancer Fund, the Cancer Councils of New South Wales, Victoria, Tasmania and South Australia, and the Cancer Foundation of Western Australia. The MARIE study was supported by the Deutsche Krebshilfe e.V. [70-2892-BR I, 106332, 108253, 108419], the Hamburg Cancer Society, the German Cancer Research Center (DKFZ) and the Federal Ministry of Education and Research (BMBF) Germany [01KH0402]. The MCBCS was supported by an NIH Specialized Program of Research Excellence (SPORE) in Breast Cancer [CA116201], the Breast Cancer Research Foundation, the Mayo Clinic Breast Cancer Registry and a generous gift from the David F. and Margaret T. Grohne Family Foundation and the Ting Tsung and Wei Fong Chao Foundation. ORIGO authors thank E. Krol-Warmerdam, and J. Blom; The contributing studies were funded by grants from the Dutch Cancer Society (UL1997-1505) and the Biobanking and Biomolecular Resources Research Infrastructure (BBMRI-NL CP16). PBCS was funded by Intramural Research Funds of the National Cancer Institute, Department of Health and Human Services, USA. The RBCS was funded by the Dutch Cancer Society (DDHK 2004-3124, DDHK 2009-4318). SEARCH is funded by programme grant from Cancer Research UK [C490/A10124. C490/A16561] and supported by the UK National Institute for Health Research Biomedical Research Centre at the University of Cambridge. Part of this work was supported by the European Community's Seventh Framework Programme under grant agreement number 223175 (grant number HEALTH-F2-2009223175) (COGS). The UKBGS is funded by Breakthrough Breast Cancer and the Institute of Cancer Research (ICR), London. ICR acknowledges NHS funding to the NIHR Biomedical Research Centre. We acknowledge funds from Breakthrough Breast Cancer, UK, in support of MGC at the time this work was carried out and funds from the Cancer Research, UK, in support of MA.
Publisher Copyright:
© 2016 The Authors The Journal of Pathology: Clinical Research published by The Pathological Society of Great Britain and Ireland and John Wiley & Sons Ltd
Keywords
- automated algorithm
- breast cancer
- immunohistochemistry
- Ki67
- tissue microarrays
ASJC Scopus subject areas
- Pathology and Forensic Medicine
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- High-throughput automated scoring of Ki67 in breast cancer tissue microarrays from the Breast Cancer Association Consortium
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