title: Investigating Diagnostic Problems of CIN1 and CIN2 Associated With High-risk HPV by Combining the Novel Molecular Biomarker PanHPVE4 With P16INK4a.
abstract: Grading cervical intraepithelial neoplasia (CIN) determines clinical management of women after abnormal cytology with potential for overdiagnosis and overtreatment. We studied a novel biomarker of human papillomavirus (HPV) life-cycle completion (panHPVE4), in combination with the minichromosome maintenance (MCM) protein cell-cycle marker and the p16INK4a transformation marker, to improve CIN diagnosis and categorization. Scoring these biomarkers alongside CIN grading by 3 pathologists was performed on 114 cervical specimens with high-risk (HR) HPV. Interobserver agreement for histopathology was moderate (κ=0.43 for CIN1/negative, 0.54 for CIN2/≤CIN1, and 0.36 for CIN3). Agreement was good or excellent for biomarker scoring (E4: κ=0.896; 95% confidence interval [CI]: 0.763-0.969; p16INK4a : κ=0.798; 95% CI: 0.712-0.884; MCM: κ=0.894; 95% CI: NC (this quantity cannot be calculated). Biomarker expression was studied by immunofluorescence and immunohistochemistry and was correlated with 104 final CIN diagnoses after histologic review. All 25 histologically negative specimens were p16INK4a and panHPVE4 negative, although 9 were MCM-positive. There were variable extents of p16INK4a positivity in 11/11 CIN1 and extensive panHPVE4 staining in 9/11. Ten CIN2 lesions expressed panHPVE4 and p16INK4a, and 13 CIN2 expressed only p16INK4a. CIN3 showed extensive p16INK4a positivity with no/minimal panHPVE4 staining. PanHPVE4, unlike MCM, distinguished CIN1 from negative. PanHPVE4 with p16INK4a separated CIN2/3 showing only expression of p16INK4a, indicating transforming HR-HPV E7 expression, from CIN1/2 showing completion of HR-HPV life cycle by E4 expression and variable p16INK4a expression. PanHPVE4 and p16INK4a staining are complementary markers that could provide simple, reliable support for diagnosing CIN. Their value in distinguishing CIN1/2 that supports HR-HPV life-cycle completion (and which might ultimately regress) from purely transforming CIN2/3 needing treatment warrants further research.
authors.affiliation: DDL Diagnostic Laboratory, Rijswijk, The Netherlands.
authors.forename: Romy
authors.initials: R
authors.lastname: van Baars
authors.affiliation: Department of Pathology, University of Cambridge, Cambridge, United Kingdom.|National Institute for Medical Research, London, United Kingdom.
authors.forename: Heather
authors.initials: H
authors.lastname: Griffin
authors.affiliation: National Institute for Medical Research, London, United Kingdom.
authors.forename: Zhonglin
authors.initials: Z
authors.lastname: Wu
authors.affiliation: National Institute for Medical Research, London, United Kingdom.
authors.forename: Yasmina Jay
authors.initials: YJ
authors.lastname: Soneji
authors.affiliation: DDL Diagnostic Laboratory, Rijswijk, The Netherlands.
authors.forename: Miekel
authors.initials: M
authors.lastname: van de Sandt
authors.affiliation: Department of cellular Pathology, University College Hospital, London, United Kingdom.
authors.forename: Rupali
authors.initials: R
authors.lastname: Arora
authors.affiliation: DDL Diagnostic Laboratory, Rijswijk, The Netherlands.
authors.forename: Jacolien
authors.initials: J
authors.lastname: van der Marel
authors.affiliation: Roosevelt Kliniek, Leiden, The Netherlands.
authors.forename: Bram
authors.initials: B
authors.lastname: Ter Harmsel
authors.affiliation: Jagiellonian University Medical College, Krakow, Poland.
authors.forename: Robert
authors.initials: R
authors.lastname: Jach
authors.affiliation: Jagiellonian University Medical College, Krakow, Poland.
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Okon
authors.affiliation: Jagiellonian University Medical College, Krakow, Poland.
authors.forename: Hubert
authors.initials: H
authors.lastname: Huras
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: Wim
authors.initials: W
authors.lastname: Quint
authors.affiliation: Department of Pathology, University of Cambridge, Cambridge, United Kingdom.|National Institute for Medical Research, London, United Kingdom.
authors.forename: John
authors.initials: J
authors.lastname: Doorbar
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D014408:Biomarkers, Tumor / Q000032:analysis*; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000032:analysis*; D005060:Europe; D005260:Female; D005455:Fluorescent Antibody Technique; D061809:Human Papillomavirus DNA Tests; D006801:Humans; D007150:Immunohistochemistry; D008875:Middle Aged; D060787:Neoplasm Grading; D015588:Observer Variation; D009856:Oncogene Proteins, Viral / Q000032:analysis*; D027383:Papillomaviridae / Q000737:chemistry* / Q000235:genetics; D030361:Papillomavirus Infections / Q000175:diagnosis* / Q000473:pathology / Q000821:virology; D011237:Predictive Value of Tests; D015203:Reproducibility of Results; D002583:Uterine Cervical Neoplasms / Q000737:chemistry / Q000175:diagnosis* / Q000473:pathology / Q000821:virology; D055815:Young Adult; D002578:Uterine Cervical Dysplasia / Q000737:chemistry / Q000175:diagnosis* / Q000473:pathology / Q000821:virology
chemical_list: D014408:Biomarkers, Tumor; D019941:Cyclin-Dependent Kinase Inhibitor p16; D009856:Oncogene Proteins, Viral