Quantitative Measurement of L1 Human Papillomavirus Type 16 Methylation for the Prediction of Preinvasive and Invasive Cervical Disease.
Methylation of the human papillomavirus (HPV) DNA has been proposed as a novel biomarker. Here, we correlated the mean methylation level of 12 CpG sites within the L1 gene, to the histological grade of cervical precancer and cancer. We assessed whether HPV L1 gene methylation can predict the presence of high-grade disease at histology in women testing positive for HPV16 genotype.



Pyrosequencing was used for DNA methylation quantification and 145 women were recruited.



We found that the L1 HPV16 mean methylation (±SD) significantly increased with disease severity (cervical intraepithelial neoplasia [CIN] 3, 17.9% [±7.2] vs CIN2, 11.6% [±6.5], P < .001 or vs CIN1, 9.0% [±3.5], P < .001). Mean methylation was a good predictor of CIN3+ cases; the area under the curve was higher for sites 5611 in the prediction of CIN2+ and higher for position 7145 for CIN3+. The evaluation of different methylation thresholds for the prediction of CIN3+ showed that the optimal balance of sensitivity and specificity (75.7% and 77.5%, respectively) and positive and negative predictive values (74.7% and 78.5%, respectively) was achieved for a methylation of 14.0% with overall accuracy of 76.7%.



Elevated methylation level is associated with increased disease severity and has good ability to discriminate HPV16-positive women that have high-grade disease or worse.
Department of Cytopathology, National and Kapodistrian University of Athens Medical School, "ATTIKON" University Hospital, Athens, Greece.
Christine
C
Kottaridi
Department of Surgery and Cancer, Institute of Reproductive and Developmental Biology, Faculty of Medicine, Imperial College London, UK.|West London Gynaecological Cancer Center, Queen Charlotte's and Chelsea, Hammersmith Hospital, Imperial Healthcare NHS Trust, London, UK.
Maria
M
Kyrgiou
Department of Cytopathology, National and Kapodistrian University of Athens Medical School, "ATTIKON" University Hospital, Athens, Greece.
Abraham
A
Pouliakis
Department of Cytopathology, National and Kapodistrian University of Athens Medical School, "ATTIKON" University Hospital, Athens, Greece.
Maria
M
Magkana
Department of Cytopathology, National and Kapodistrian University of Athens Medical School, "ATTIKON" University Hospital, Athens, Greece.
Evangelia
E
Aga
Department of Cytopathology, National and Kapodistrian University of Athens Medical School, "ATTIKON" University Hospital, Athens, Greece.
Aris
A
Spathis
Department of Surgery and Cancer, Institute of Reproductive and Developmental Biology, Faculty of Medicine, Imperial College London, UK.|West London Gynaecological Cancer Center, Queen Charlotte's and Chelsea, Hammersmith Hospital, Imperial Healthcare NHS Trust, London, UK.
Anita
A
Mitra
Third Department of Obstetrics and Gynecology, University of Athens Medical School, "ATTIKON" University Hospital, Athens.
George
G
Makris
Third Department of Obstetrics and Gynecology, University of Athens Medical School, "ATTIKON" University Hospital, Athens.
Charalampos
C
Chrelias
Second Department of Internal Medicine and Research Institute, University of Athens Medical School, "ATTIKON" University Hospital, Athens.
Vassiliki
V
Mpakou
Department of Obstetrics and Gynecology, University Hospital of Ioannina, Ioannina, Greece.
Evangelos
E
Paraskevaidis
Second Department of Pathology, University of Athens Medical School, "ATTIKON" University Hospital, Athens, Greece.
John G
JG
Panayiotides
Department of Cytopathology, National and Kapodistrian University of Athens Medical School, "ATTIKON" University Hospital, Athens, Greece.
Petros
P
Karakitsos
D000328:Adult; D000368:Aged; D036022:Capsid Proteins / Q000235:genetics*; D018899:CpG Islands; D019175:DNA Methylation; D004279:DNA, Viral / Q000737:chemistry; D005260:Female; D005838:Genotype; D006115:Greece; D052162:Human papillomavirus 16; D006801:Humans; D016014:Linear Models; D008875:Middle Aged; D009856:Oncogene Proteins, Viral / Q000235:genetics*; D030361:Papillomavirus Infections / Q000175:diagnosis*; D011446:Prospective Studies; D012680:Sensitivity and Specificity; D006113:United Kingdom; D002577:Uterine Cervical Diseases / Q000175:diagnosis* / Q000821:virology; D055815:Young Adult; D002578:Uterine Cervical Dysplasia / Q000175:diagnosis / Q000821:virology
D036022:Capsid Proteins; D004279:DNA, Viral; C114580:HPV L1 protein, Human papillomavirus; D009856:Oncogene Proteins, Viral
CIN; HPV L1 gene methylation; Human Papillomavirus; cervical intraepithelial neoplasia; pyrosequencing