title: High-throughput automated scoring of Ki67 in breast cancer tissue microarrays from the Breast Cancer Association Consortium.
abstract: Automated methods are needed to facilitate high-throughput and reproducible scoring of Ki67 and other markers in breast cancer tissue microarrays (TMAs) in large-scale studies. To address this need, we developed an automated protocol for Ki67 scoring and evaluated its performance in studies from the Breast Cancer Association Consortium. We utilized 166 TMAs containing 16,953 tumour cores representing 9,059 breast cancer cases, from 13 studies, with information on other clinical and pathological characteristics. TMAs were stained for Ki67 using standard immunohistochemical procedures, and scanned and digitized using the Ariol system. An automated algorithm was developed for the scoring of Ki67, and scores were compared to computer assisted visual (CAV) scores in a subset of 15 TMAs in a training set. We also assessed the correlation between automated Ki67 scores and other clinical and pathological characteristics. Overall, we observed good discriminatory accuracy (AUC = 85%) and good agreement (kappa = 0.64) between the automated and CAV scoring methods in the training set. The performance of the automated method varied by TMA (kappa range= 0.37-0.87) and study (kappa range = 0.39-0.69). The automated method performed better in satisfactory cores (kappa = 0.68) than suboptimal (kappa = 0.51) cores (p-value for comparison = 0.005); and among cores with higher total nuclei counted by the machine (4,000-4,500 cells: kappa = 0.78) than those with lower counts (50-500 cells: kappa = 0.41; p-value = 0.010). Among the 9,059 cases in this study, the correlations between automated Ki67 and clinical and pathological characteristics were found to be in the expected directions. Our findings indicate that automated scoring of Ki67 can be an efficient method to obtain good quality data across large numbers of TMAs from multicentre studies. However, robust algorithm development and rigorous pre- and post-analytical quality control procedures are necessary in order to ensure satisfactory performance.
authors: Division of Genetics and Epidemiology The Institute of Cancer Research London UK.
authors: Mustapha
authors: M
authors: Abubakar
authors: Cancer Research UK Cambridge Institute, University of Cambridge Cambridge UK.
authors: William J
authors: WJ
authors: Howat
authors: Breakthrough Breast Cancer Research Centre, Division of Breast Cancer Research, The Institute of Cancer Research London UK.
authors: Frances
authors: F
authors: Daley
authors: Academic Department of Biochemistry, Royal Marsden Hospital Fulham Road London.
authors: Lila
authors: L
authors: Zabaglo
authors: Cancer Research UK Cambridge Institute, University of Cambridge Cambridge UK.
authors: Leigh-Anne
authors: LA
authors: McDuffus
authors: Centre for Cancer Genetic Epidemiology, Department of Oncology, University of Cambridge Cambridge UK.
authors: Fiona
authors: F
authors: Blows
authors: Division of Genetics and Epidemiology The Institute of Cancer Research London UK.
authors: Penny
authors: P
authors: Coulson
authors: Cancer Research UK Cambridge Institute, University of Cambridge Cambridge UK.
authors: H
authors: H
authors: Raza Ali
authors: Human Genetics Group, Human Cancer Genetics Program, Spanish National Cancer Research Centre (CNIO)MadridSpain; Centro de Investigacion en Red de Enfermedades Raras (CIBERER)ValenciaSpain.
authors: Javier
authors: J
authors: Benitez
authors: Cancer Epidemiology Centre, Cancer Council VictoriaMelbourneAustralia; Centre for Epidemiology and Biostatistics, Melbourne School of Population and Global health, The University of MelbourneMelbourneAustralia.
authors: Roger
authors: R
authors: Milne
authors: Division of Clinical Epidemiology and Aging Research, German Cancer Research Center (DKFZ)HeidelbergGermany; Division of Preventive Oncology, German Cancer Research Center (DKFZ), and National Center for Tumor Diseases (NCT)HeidelbergGermany; German Cancer Consortium (DKTK), German Cancer Research Center (DKFZ)HeidelbergGermany.
authors: Herman
authors: H
authors: Brenner
authors: Saarland Cancer Registry Saarland Germany.
authors: Christa
authors: C
authors: Stegmaier
authors: School of Medicine, Institute of Clinical Medicine, Pathology and Forensic Medicine, Cancer Center of Eastern Finland, University of Eastern FinlandKuopioFinland; Imaging Center, Department of Clinical Pathology, Kuopio University HospitalKuopioFinland.
authors: Arto
authors: A
authors: Mannermaa
authors: Division of Cancer Epidemiology, German Cancer Research Center (DKFZ)HeidelbergGermany; University Cancer Center Hamburg (UCCH), University Medical Center Hamburg-EppendorfHamburgGermany.
authors: Jenny
authors: J
authors: Chang-Claude
authors: Division of Cancer Epidemiology, German Cancer Research Center (DKFZ) Heidelberg Germany.
authors: Anja
authors: A
authors: Rudolph
authors: Department of Pathology Institute of Pathology, Heidelberg University Hospital Germany.
authors: Peter
authors: P
authors: Sinn
authors: Department of Laboratory Medicine and Pathology Mayo Clinic Rochester, MN USA.
authors: Fergus J
authors: FJ
authors: Couch
authors: Department of Surgery Leiden University Medical Center The Netherlands.
authors: Rob A E M
authors: RA
authors: Tollenaar
authors: Department of Human Genetics & Department of Pathology Leiden University Medical Center Leiden The Netherlands.
authors: Peter
authors: P
authors: Devilee
authors: Usher Institute of Population Health Sciences and Informatics, The University of Edinburgh Scotland UK.
authors: Jonine
authors: J
authors: Figueroa
authors: Division of Cancer Epidemiology and Genetics National Cancer Institute Rockville Maryland USA.
authors: Mark E
authors: ME
authors: Sherman
authors: Department of Cancer Epidemiology and Prevention M. Sklodowska-Curie Memorial Cancer Center and Institute of Oncology Warsaw Poland.
authors: Jolanta
authors: J
authors: Lissowska
authors: Laboratory of Pathology National Cancer Institute, National Institutes of Health Rockville MD USA.
authors: Stephen
authors: S
authors: Hewitt
authors: Faculty of Medicine Academic Unit of Cancer Sciences Southampton General Hospital Southampton UK.
authors: Diana
authors: D
authors: Eccles
authors: Family Cancer Clinic, Department of Medical Oncology, Erasmus MC Cancer Institute Rotterdam The Netherlands.
authors: Maartje J
authors: MJ
authors: Hooning
authors: Family Cancer Clinic, Department of Medical Oncology, Erasmus MC Cancer Institute Rotterdam The Netherlands.
authors: Antoinette
authors: A
authors: Hollestelle
authors: Family Cancer Clinic, Department of Medical Oncology, Erasmus MC Cancer Institute Rotterdam The Netherlands.
authors: John
authors: J
authors: Wm Martens
authors: Department of Pathology Erasmus MC Cancer Institute Rotterdam The Netherlands.
authors: Carolien
authors: C
authors: Hm van Deurzen
authors: Department of Genetics QIMR Berghofer Medical Research Institute Brisbane Australia.
authors: Centre for Cancer Genetic Epidemiology, Department of Public Health and Primary Care, University of Cambridge Cambridge UK.
authors: Manjeet K
authors: MK
authors: Bolla
authors: Centre for Cancer Genetic Epidemiology, Department of Public Health and Primary Care, University of Cambridge Cambridge UK.
authors: Qin
authors: Q
authors: Wang
authors: Division of Genetics and Epidemiology The Institute of Cancer Research London UK.
authors: Michael
authors: M
authors: Jones
authors: Division of Genetics and Epidemiology The Institute of Cancer Research London UK.
authors: Minouk
authors: M
authors: Schoemaker
authors: Division of Molecular Pathology Netherlands Cancer Institute, Antoni van Leeuwenhoek Hospital Amsterdam The Netherlands.
authors: Annegien
authors: A
authors: Broeks
authors: Division of Psychosocial Research and Epidemiology Netherlands Cancer Institute, Antoni van Leeuwenhoek Hospital Amsterdam The Netherlands.
authors: Flora E
authors: FE
authors: van Leeuwen
authors: Division of Molecular Pathology Netherlands Cancer Institute, Antoni van Leeuwenhoek Hospital Amsterdam The Netherlands.
authors: Laura
authors: L
authors: Van't Veer
authors: Division of Genetics and EpidemiologyThe Institute of Cancer ResearchLondonUK; Division of Breast Cancer ResearchThe Institute of Cancer ResearchLondonUK.
authors: Anthony J
authors: AJ
authors: Swerdlow
authors: Breakthrough Breast Cancer Research Centre, Division of Breast Cancer Research, The Institute of Cancer Research London UK.
authors: Nick
authors: N
authors: Orr
authors: Breakthrough Breast Cancer Research Centre, Division of Breast Cancer Research, The Institute of Cancer ResearchLondonUK; Academic Department of Biochemistry, Royal Marsden HospitalFulham RoadLondon.
authors: Mitch
authors: M
authors: Dowsett
authors: Centre for Cancer Genetic Epidemiology, Department of Oncology, University of CambridgeCambridgeUK; Centre for Cancer Genetic Epidemiology, Department of Public Health and Primary Care, University of CambridgeCambridgeUK.
authors: Douglas
authors: D
authors: Easton
authors: Division of Molecular PathologyNetherlands Cancer Institute, Antoni van Leeuwenhoek HospitalAmsterdamThe Netherlands; Division of Psychosocial Research and EpidemiologyNetherlands Cancer Institute, Antoni van Leeuwenhoek HospitalAmsterdamThe Netherlands.
authors: Marjanka K
authors: MK
authors: Schmidt
authors: Centre for Cancer Genetic Epidemiology, Department of Oncology, University of CambridgeCambridgeUK; Centre for Cancer Genetic Epidemiology, Department of Public Health and Primary Care, University of CambridgeCambridgeUK.
authors: Paul D
authors: PD
authors: Pharoah
authors: Division of Cancer Epidemiology and Genetics National Cancer Institute Rockville Maryland USA.
authors: Montserrat
authors: M
authors: Garcia-Closas
keywords: Ki67; automated algorithm; breast cancer; immunohistochemistry; tissue microarrays