P16INK4A flow cytometry of exfoliated cervical cells: Its role in quantitative pathology and clinical diagnosis of squamous intraepithelial lesions. Issue 3 (7th March 2023)

• Record Type: Journal Article
• Authors: He, Yifeng; Shi, Jun; Zhao, Hui; Wang, Yuefei; Zhang, Chi; Han, Sai; He, Qizhi; Li, Xiaolan; Li, Shangji; Wang, Wenjing; Yi, Muhua; Hu, Xiaoling; Xing, Zhihua; Han, Hao; Gao, Yinshuang; Zhou, Qing; Lu, Linlin; Guo, Jianfen; Cao, Hui; Lu, Caiping; Hou, Yanqiang; Chen, Dan; Yang, Fengyun; Lei, Ping; Di, Wen; Qian, Ji; Xia, Yi; Zhang, Youzhong; Deng, Yang; Zhu, Jianlong; Xu, Congjian;
• Is Part Of: Clinical and translational medicine. Volume 13:Issue 3(2023)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2023-03-07
• Subjects: cervical cancer -- flow cytometry -- human papillomavirus -- immunostaining -- p16INK4A -- papanicolaou smear -- pathology -- quantification -- squamous intraepithelial lesion; Clinical medicine -- Periodicals; Medicine, Experimental -- Periodicals; Medical innovations -- Periodicals; Molecular biology -- Periodicals; Pathology, Molecular -- Periodicals; 616.027
• DOI: 10.1002/ctm2.1209
• Languages: English
• ISSNs: 2001-1326

Abstract

Abstract: Background: P16 INK4A is a surrogate signature compensating for the specificity and/or sensitivity deficiencies of the human papillomavirus (HPV) DNA and Papanicolaou smear (Pap) co‐test for detecting high‐grade cervical squamous intraepithelial lesions or worse (HSIL+). However, traditional p16INK4A immunostaining is labour intensive and skill demanding, and subjective biases cannot be avoided. Herein, we created a high‐throughput, quantitative diagnostic device, p16INK4A flow cytometry (FCM) and assessed its performances in cervical cancer screening and prevention. Methods: P16 INK4A FCM was built upon a novel antibody clone and a series of positive and negative (p16 INK4A ‐knockout) standards. Since 2018, 24 100‐women (HPV‐positive/‐negative, Pap‐normal/‐abnormal) have been enrolled nationwide for two‐tier validation work. In cross‐sectional studies, age‐ and viral genotype‐dependent expression of p16 INK4A was investigated, and optimal diagnostic parameter cut‐offs (using colposcopy and biopsy as a gold standard) were obtained. In cohort studies, the 2‐year prognostic values of p16 INK4A were investigated with other risk factors by multivariate regression analyses in three cervicopathological conditions: HPV‐positive Pap‐normal, Pap‐abnormal biopsy‐negative and biopsy‐confirmed LSIL. Results: P16 INK4A FCM detected a minimal ratio of 0.01% positive cells. The p16 INK4A ‐positive ratio was 13.9 ± 1.8% among HPV‐negative NILM women and peaked at the ages of 40–49 years; after HPV infection, the ratio increased to 15.1 ± 1.6%, varying with the carcinogenesis of the viral genotype. Further increments were found in women with neoplastic lesions (HPV‐negative: 17.7 ± 5.0–21.4 ± 7.2%; HPV‐positive: 18.0 ± 5.2–20.0 ± 9.9%). Extremely low expression of p16 INK4A was observed in women with HSILs. As the HPV‐combined double‐cut‐off‐ratio criterion was adopted, a Youden's index of 0.78 was obtained, which was significantly higher than that (0.72) of the HPV and Pap co‐test. The p16 INK4A ‐abnormal situation was an independent HSIL+ risk factor for 2‐year outcomes in all three cervicopathological conditions investigated (hazard ratios: 4.3–7.2). Conclusions: FCM‐based p16 INK4A quantification offers a better choice for conveniently and precisely monitoring the occurrence of HSIL+ and directing risk‐stratification‐based interventions. Abstract : The quantification detection system—p16 INK4A FCM was established for triaging cervical high‐grade neoplasia/cancer. 24 100‐women cross‐sectional studies revealed that HSIL+‐triaging efficacy of HPV and p16 INK4A FCM co‐test is significantly higher than that of HPV and Pap co‐test. 2‐year cohort studies indicated that HPV‐positive/Pap‐abnormal p16 INK4A ‐abnormal women would run a HSIL+‐risk > 4‐fold higher than those with a p16 INK4A ‐normal level.