title: Performance of automated scoring of ER, PR, HER2, CK5/6 and EGFR in breast cancer tissue microarrays in the Breast Cancer Association Consortium.
abstract: Breast cancer risk factors and clinical outcomes vary by tumour marker expression. However, individual studies often lack the power required to assess these relationships, and large-scale analyses are limited by the need for high throughput, standardized scoring methods. To address these limitations, we assessed whether automated image analysis of immunohistochemically stained tissue microarrays can permit rapid, standardized scoring of tumour markers from multiple studies. Tissue microarray sections prepared in nine studies containing 20 263 cores from 8267 breast cancers stained for two nuclear (oestrogen receptor, progesterone receptor), two membranous (human epidermal growth factor receptor 2 and epidermal growth factor receptor) and one cytoplasmic (cytokeratin 5/6) marker were scanned as digital images. Automated algorithms were used to score markers in tumour cells using the Ariol system. We compared automated scores against visual reads, and their associations with breast cancer survival. Approximately 65-70% of tissue microarray cores were satisfactory for scoring. Among satisfactory cores, agreement between dichotomous automated and visual scores was highest for oestrogen receptor (Kappa = 0.76), followed by human epidermal growth factor receptor 2 (Kappa = 0.69) and progesterone receptor (Kappa = 0.67). Automated quantitative scores for these markers were associated with hazard ratios for breast cancer mortality in a dose-response manner. Considering visual scores of epidermal growth factor receptor or cytokeratin 5/6 as the reference, automated scoring achieved excellent negative predictive value (96-98%), but yielded many false positives (positive predictive value = 30-32%). For all markers, we observed substantial heterogeneity in automated scoring performance across tissue microarrays. Automated analysis is a potentially useful tool for large-scale, quantitative scoring of immunohistochemically stained tissue microarrays available in consortia. However, continued optimization, rigorous marker-specific quality control measures and standardization of tissue microarray designs, staining and scoring protocols is needed to enhance results.
authors: Cancer Research UK Cambridge Institute, University of Cambridge Cambridge UK.
authors: William J
authors: WJ
authors: Howat
authors: Centre for Cancer Genetic Epidemiology, Department of Oncology University of Cambridge Cambridge UK.
authors: Fiona M
authors: FM
authors: Blows
authors: Breast Pathology Addenbrookes Hospital Cambridge UK.
authors: Elena
authors: E
authors: Provenzano
authors: Division of Genetics and Epidemiology The Institute of Cancer Research London UK.
authors: Mark N
authors: MN
authors: Brook
authors: Cancer Research UK Cambridge Institute, University of CambridgeCambridgeUK; Department of OncologyUniversity of CambridgeCambridgeUK.
authors: Lorna
authors: L
authors: Morris
authors: Breakthrough Breast Cancer Research Unit, Division of Cancer Studies King's College London, Guy's Hospital London UK.
authors: Patrycja
authors: P
authors: Gazinska
authors: Cancer Research UK Cambridge Institute, University of Cambridge Cambridge UK.
authors: Nicola
authors: N
authors: Johnson
authors: Cancer Research UK Cambridge Institute, University of Cambridge Cambridge UK.
authors: Leigh-Anne
authors: LA
authors: McDuffus
authors: Cancer Research UK Cambridge Institute, University of Cambridge Cambridge UK.
authors: Jodi
authors: J
authors: Miller
authors: Division of Cancer Studies, NIHR Comprehensive Biomedical Research Centre Guy's & St. Thomas' NHS Foundation Trust in partnership with King's College London London UK.
authors: Elinor J
authors: EJ
authors: Sawyer
authors: Research Oncology, Division of Cancer Studies King's College London, Guy's Hospital London UK.
authors: Sarah
authors: S
authors: Pinder
authors: Department of Pathology Erasmus University Medical Center Rotterdam The Netherlands.
authors: Carolien H M
authors: CH
authors: van Deurzen
authors: Centre for Tumour BiologyBarts Institute of CancerBartsUK; The London School of Medicine and DentistryLondonUK.
authors: Louise
authors: L
authors: Jones
authors: School of Medicine, Institute of Clinical Medicine, Pathology and Forensic MedicineCancer Center of Eastern Finland, University of Eastern FinlandKuopioFinland; Imaging Center, Department of Clinical PathologyKuopio University HospitalKuopioFinland.
authors: Reijo
authors: R
authors: Sironen
authors: Department of Laboratory Medicine and Pathology Mayo Clinic Rochester MN USA.
authors: Daniel
authors: D
authors: Visscher
authors: Cancer Research UK Cambridge Institute, University of Cambridge Cambridge UK.
authors: Carlos
authors: C
authors: Caldas
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: Division of Genetics and Epidemiology The Institute of Cancer Research London UK.
authors: Penny
authors: P
authors: Coulson
authors: Core Facility for Molecular Pathology and Biobanking Netherlands Cancer Institute, Antoni van Leeuwenhoek Hospital Amsterdam The Netherlands.
authors: Annegien
authors: A
authors: Broeks
authors: Department of Pathology, Division of Diagnostic Oncology Netherlands Cancer Institute, Antoni van Leeuwenhoek Hospital Amsterdam The Netherlands.
authors: Joyce
authors: J
authors: Sanders
authors: Department of Pathology, Division of Diagnostic Oncology Netherlands Cancer Institute, Antoni van Leeuwenhoek Hospital Amsterdam The Netherlands.
authors: Jelle
authors: J
authors: Wesseling
authors: Department of Obstetrics and Gynecology University of Helsinki and Helsinki University Central Hospital Helsinki Finland.
authors: Heli
authors: H
authors: Nevanlinna
authors: Department of Obstetrics and Gynecology University of Helsinki and Helsinki University Central Hospital Helsinki Finland.
authors: Rainer
authors: R
authors: Fagerholm
authors: Department of Oncology Helsinki University Central Hospital Helsinki Finland.
authors: Carl
authors: C
authors: Blomqvist
authors: Department of Pathology Helsinki University Central Hospital Helsinki Finland.
authors: Päivi
authors: P
authors: Heikkilä
authors: Cancer Research UK Cambridge Institute, University of Cambridge Cambridge UK.
authors: H Raza
authors: HR
authors: Ali
authors: Cancer Research UK Cambridge Institute, University of Cambridge Cambridge UK.
authors: Sarah-Jane
authors: SJ
authors: Dawson
authors: Division of Cancer Epidemiology and Genetics National Cancer Institute Rockville Maryland USA.
authors: Jonine
authors: J
authors: Figueroa
authors: Department of Cancer Epidemiology and Prevention M. Sklodowska-Curie Memorial Cancer Center & Institute of Oncology Warsaw Poland.
authors: Jolanta
authors: J
authors: Lissowska
authors: Division of Cancer Epidemiology and Genetics National Cancer Institute Rockville Maryland USA.
authors: Louise
authors: L
authors: Brinton
authors: School of Medicine, Institute of Clinical Medicine, Pathology and Forensic MedicineCancer Center of Eastern Finland, University of Eastern FinlandKuopioFinland; Imaging Center, Department of Clinical PathologyKuopio University HospitalKuopioFinland.
authors: Arto
authors: A
authors: Mannermaa
authors: Kuopio University Hospital, Cancer CenterKuopioFinland; School of Medicine, Institute of Clinical MedicineUniversity of Eastern Finland, Oncology and Central Hospital of Central Finland, Central Finland Hospital DistrictKuopioFinland.
authors: Vesa
authors: V
authors: Kataja
authors: School of Medicine, Institute of Clinical Medicine, Pathology and Forensic MedicineCancer Center of Eastern Finland, University of Eastern FinlandKuopioFinland; Imaging Center, Department of Clinical PathologyKuopio University HospitalKuopioFinland.
authors: Veli-Matti
authors: VM
authors: Kosma
authors: CRUK/YCR Sheffield Cancer Research Centre, Department of Oncology University of Sheffield Sheffield UK.
authors: Angela
authors: A
authors: Cox
authors: CRUK/YCR Sheffield Cancer Research Centre, Department of Oncology University of Sheffield Sheffield UK.
authors: Ian W
authors: IW
authors: Brock
authors: Academic Unit of Pathology, Department of Neuroscience University of Sheffield Sheffield UK.
authors: Simon S
authors: SS
authors: Cross
authors: CRUK/YCR Sheffield Cancer Research Centre, Department of Oncology University of Sheffield Sheffield UK.
authors: Malcolm W
authors: MW
authors: Reed
authors: Department of Laboratory Medicine and Pathology Mayo Clinic Rochester MN USA.
authors: Fergus J
authors: FJ
authors: Couch
authors: Department of Health Sciences Research Mayo Clinic Rochester MN USA.
authors: Janet E
authors: JE
authors: Olson
authors: Department of Human Genetics & Department of Pathology Leiden University Medical Center Leiden The Netherlands.
authors: Peter
authors: P
authors: Devillee
authors: Department of Surgical Oncology Leiden University Medical Center RC Leiden The Netherlands.
authors: Wilma E
authors: WE
authors: Mesker
authors: Family Cancer Clinic, Department of Medical Oncology Erasmus MC Cancer Institute Rotterdam The Netherlands.
authors: Caroline M
authors: CM
authors: Seyaneve
authors: Family Cancer Clinic, Department of Medical Oncology Erasmus MC Cancer Institute Rotterdam The Netherlands.
authors: Antoinette
authors: A
authors: Hollestelle
authors: Human Genetics Group, Human Cancer Genetics ProgramSpanish National Cancer Research Centre (CNIO)MadridSpain; Centro de Investigación en Red de Enfermedades Raras (CIBERER)ValenciaSpain.
authors: Javier
authors: J
authors: Benitez
authors: Servicio de Cirugía General y Especialidades Hospital Monte Naranco Oviedo Spain.
authors: Jose Ignacio Arias
authors: JI
authors: Perez
authors: Servicio de Anatomía Patológica Hospital Monte Naranco Oviedo Spain.
authors: Primitiva
authors: P
authors: Menéndez
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 OncologyUniversity of CambridgeCambridgeUK; Centre for Cancer Genetic Epidemiology, Department of Public Health and Primary CareUniversity of CambridgeCambridgeUK.
authors: Douglas F
authors: DF
authors: Easton
authors: Division of Molecular Pathology Netherlands Cancer Institute, Antoni van Leeuwenhoek Hospital Amsterdam The Netherlands.
authors: Marjanka K
authors: MK
authors: Schmidt
authors: Centre for Cancer Genetic Epidemiology, Department of OncologyUniversity of CambridgeCambridgeUK; Centre for Cancer Genetic Epidemiology, Department of Public Health and Primary CareUniversity of CambridgeCambridgeUK.
authors: Paul D
authors: PD
authors: Pharoah
authors: Division of Cancer Epidemiology and Genetics National Cancer Institute Rockville Maryland USA.
authors: Mark E
authors: ME
authors: Sherman
authors: Division of Genetics and EpidemiologyThe Institute of Cancer ResearchLondonUK; Breakthrough Breast Cancer Research Centre, Division of Breast Cancer ResearchThe Institute of Cancer ResearchLondonUK.
authors: Montserrat
authors: M
authors: García-Closas
keywords: automated scoring; breast tumours; digital pathology; immunohistochemistry; tissue microarrays