title: Stratification of HPV-induced cervical pathology using the virally encoded molecular marker E4 in combination with p16 or MCM.
abstract: High-risk human papillomavirus (HPV) types cause cervical lesions of varying severity, ranging from transient productive infections to high-grade neoplasia. Disease stratification requires the examination of lesional pathology, and possibly also the detection of biomarkers. P16(INK4a) and MCM are established surrogates of high-risk HPV E6/E7 activity, and can be extensively expressed in high-grade lesions. Here we have combined these two cellular biomarkers with detection of the abundant HPV-encoded E4 protein in order to identify both productive and transforming lesions. This approach has allowed us to distinguish true papillomavirus infections from similar pathologies, and has allowed us to divide the heterogeneous CIN2 category into those that are CIN1-like and express E4, and those that more closely resemble nonproductive CIN3. To achieve this, 530 lesional areas were evaluated according to standard pathology criteria and by using a multiple staining approach that allows us to superimpose biomarker patterns either singly or in combination onto an annotated hematoxylin and eosin (H&E) image. Conventional grading of neoplasia was established by review panel, and compared directly with the composite molecular pathology visualized on the same tissue section. The detection of E4 coincided with the onset of vacuolation, becoming abundant in koilocytes as the MCM marker declined and cells lost their defined nuclear margins as visualized by standard H&E staining. Of the dual marker approaches, p16(INK4a) and E4 appeared most promising, with E4 generally identifying areas of low-grade disease even when p16(INK4a) was present. Extensive p16(INK4a) expression usually coincided with an absence of E4 expression or its focal retention in sporadic cells within the lesion. Our results suggest that a straightforward molecular evaluation of HPV life-cycle deregulation in cervical neoplasia may help improve disease stratification, and that this can be achieved using complementary molecular biomarker pairs such as MCM/E4 or, more promisingly, p16(INK4a)/E4 as an adjunct to conventional pathology.
authors.affiliation: 1] Department of Pathology, University of Cambridge, Cambridge, UK [2] National Institute for Medical Research, London, UK.
authors.forename: Heather
authors.initials: H
authors.lastname: Griffin
authors.affiliation: National Institute for Medical Research, London, UK.
authors.forename: Yasmina
authors.initials: Y
authors.lastname: Soneji
authors.affiliation: DDL Diagnostic Laboratory, Rijswijk, The Netherlands.
authors.forename: Romy
authors.initials: R
authors.lastname: Van Baars
authors.affiliation: University College Hospital, London, UK.
authors.forename: Rupali
authors.initials: R
authors.lastname: Arora
authors.affiliation: DDL Diagnostic Laboratory, Rijswijk, The Netherlands.
authors.forename: David
authors.initials: D
authors.lastname: Jenkins
authors.affiliation: DDL Diagnostic Laboratory, Rijswijk, The Netherlands.
authors.forename: Miekel
authors.initials: M
authors.lastname: van de Sandt
authors.affiliation: National Institute for Medical Research, London, UK.
authors.forename: Zhonglin
authors.initials: Z
authors.lastname: Wu
authors.affiliation: DDL Diagnostic Laboratory, Rijswijk, The Netherlands.
authors.forename: Wim
authors.initials: W
authors.lastname: Quint
authors.affiliation: Department of Gynecology and Oncology, Jagiellonian University College, Krakow, Poland.
authors.forename: Robert
authors.initials: R
authors.lastname: Jach
authors.affiliation: Department of Gynecology and Oncology, Jagiellonian University College, Krakow, Poland.
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Okon
authors.affiliation: Department of Gynecology and Oncology, Jagiellonian University College, Krakow, Poland.
authors.forename: Hubert
authors.initials: H
authors.lastname: Huras
authors.affiliation: University College Hospital, London, UK.
authors.forename: Albert
authors.initials: A
authors.lastname: Singer
authors.affiliation: 1] Department of Pathology, University of Cambridge, Cambridge, UK [2] National Institute for Medical Research, London, UK.
authors.forename: John
authors.initials: J
authors.lastname: Doorbar
mesh_terms: D014408:Biomarkers, Tumor / Q000378:metabolism; D002584:Cervix Uteri / Q000473:pathology* / Q000821:virology; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000378:metabolism*; D005260:Female; D006801:Humans; D060787:Neoplasm Grading; D009856:Oncogene Proteins, Viral / Q000378:metabolism*; D027383:Papillomaviridae; D030361:Papillomavirus Infections / Q000473:pathology* / Q000821:virology; D002583:Uterine Cervical Neoplasms / Q000473:pathology* / Q000821:virology; D002578:Uterine Cervical Dysplasia / Q000473:pathology* / Q000821:virology
chemical_list: D014408:Biomarkers, Tumor; D019941:Cyclin-Dependent Kinase Inhibitor p16; D009856:Oncogene Proteins, Viral; C069119:oncogene protein E4, Human papillomavirus type 16