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HPV genotype-specific p16/Ki67 expression with machine learning-assisted assessment in cervical neoplasia
Meryem Kececi Oguzhanoglu1, Esen Gul Uzuner2, Kursat Oguzhanoglu1
1Department of Obstetrics and Gynecology, Haseki Training and Research Hospital, University of Health Sciences, Istanbul, Türkiye.
Introduction:
Genotype-specific patterns of dual p16/Ki67 immunoexpression and their integration with computational assessment remain insufficiently delineated in cervical neoplasia. The present investigation characterized biomarker expression stratified by HPV genotype and evaluated the methodological feasibility of a deep learning-assisted scoring pipeline.
Methods:
A single-center cross-sectional investigation was conducted on 100 HPV-positive women, stratified into three categories: HPV16 mono-infection (n=33), non-HPV16 high-risk mono-infection (n=33), and multi-genotype co-infection (n=34). Whole-slide p16/Ki67 immunohistochemistry was scored through real-time consensus by two pathologists of differing experience levels, blinded to HPV genotype. A ResNet50-based computational pipeline was developed as a methodological feasibility demonstration and evaluated on an independent held-out test set (n=25). Between-group comparisons were performed using the Mann-Whitney U test with Bonferroni correction; multivariable logistic regression and receiver operating characteristic (ROC) analysis were employed to quantify biomarker discriminative performance; and analysis of covariance (ANCOVA) with age as a continuous covariate was undertaken as a pre-specified sensitivity analysis.
Results:
HPV16 mono-infection exhibited significantly elevated p16 immunoexpression (52.4 ± 27.6%) relative to non-HPV16 high-risk genotypes (31.1 ± 22.8%; p = 0.0021; Cohen's d = 0.844) and co-infections (34.6 ± 24.1%); between-group differences persisted following age adjustment. On the independent test set, the computational model yielded an accuracy of 96.0% (95% CI, 78.3-99.9%) and an AUC-ROC of 0.96. Given the restricted test set dimension and single-center design, these metrics should be construed as preliminary.
Conclusions:
HPV16 mono-infection is associated with distinctly elevated dual p16/Ki67 immunoexpression, providing methodological support for genotype-informed cytological risk stratification. The computational pipeline demonstrates technical feasibility; however, external multicenter validation is required prior to any consideration of clinical implementation.
