Amsterdam UMC, Vrije Universiteit Amsterdam, Pathology, Cancer Center Amsterdam, de Boelelaan 1117, 1081 HV, Amsterdam, The Netherlands.
Department of Obstetrics and Gynecology, University of Pretoria, Pretoria, South Africa.
Mod Pathol. 2020 Oct;33(10):1968-1978. doi: 10.1038/s41379-020-0528-x. Epub 2020 Apr 6.
This study aims to characterize cervical intraepithelial neoplasia (CIN) in women living with HIV using biomarkers. Immunohistochemical (IHC) staining for human papillomavirus (HPV) E4 protein indicates CIN with productive HPV infection, whereas Ki-67 and p16 indicate CIN with transforming characteristics, which may be further characterized using DNA hypermethylation, indicative for advanced transforming CIN. Cervical biopsies (n = 175) from 102 HPV positive women living with HIV were independently reviewed by three expert pathologists. The consensus CIN grade was used as reference standard. IHC staining patterns were scored for Ki-67 (0-3), p16 (0-3), and E4 (0-2) and correlated to methylation levels of four cellular genes in corresponding cervical scrapes. Reference standards and immunoscores were obtained from 165 biopsies:15 no dysplasia, 91 CIN1, 31 CIN2, and 28 CIN3. Ki-67 and p16 scores increased with increasing CIN grade, while E4 positivity was highest in CIN1 and CIN2 lesions. E4 positive CIN1 lesions had higher Ki-67 and p16 scores and higher methylation levels compared with E4 negative CIN1 lesions. E4 positive biopsies with low cumulative Ki-67/p16 immunoscores (0-3) had significantly higher methylation levels compared with E4 negative biopsies. No significant differences in Ki-67 and p16 scores and methylation levels were observed between E4 negative and positive CIN2 or CIN3 lesions. The presence of high methylation levels in scrapes of CIN lesions with IHC characteristics of both productive (E4 positive) and transforming infections (increased Ki-67/p16 expression) in women living with HIV might indicate a rapid aggressive course of HPV infections towards cancer in these women.
本研究旨在利用生物标志物对 HIV 感染者的宫颈上皮内瘤变（CIN）进行特征描述。人乳头瘤病毒（HPV）E4 蛋白的免疫组化（IHC）染色可指示具有活跃 HPV 感染的 CIN，而 Ki-67 和 p16 则指示具有转化特征的 CIN，这可以进一步通过 DNA 超甲基化来表征，提示高级转化的 CIN。从 102 名 HPV 阳性的 HIV 感染者中采集了 175 份宫颈活检组织，由 3 位专家病理学家进行独立复查。共识 CIN 分级被用作参考标准。对 Ki-67（0-3）、p16（0-3）和 E4（0-2）的 IHC 染色模式进行评分，并与相应宫颈刮片中的四个细胞基因的甲基化水平相关联。从 165 份活检组织中获得参考标准和免疫评分：15 份无发育不良，91 份 CIN1，31 份 CIN2 和 28 份 CIN3。Ki-67 和 p16 评分随 CIN 分级的增加而增加，而 E4 阳性在 CIN1 和 CIN2 病变中最高。E4 阳性的 CIN1 病变的 Ki-67 和 p16 评分以及甲基化水平均高于 E4 阴性的 CIN1 病变。E4 阳性且累积 Ki-67/p16 免疫评分（0-3）较低的活检组织的甲基化水平显著高于 E4 阴性的活检组织。E4 阴性和阳性的 CIN2 或 CIN3 病变之间，Ki-67 和 p16 评分以及甲基化水平均无显著差异。在 HIV 感染者中，具有活跃（E4 阳性）和转化感染（Ki-67/p16 表达增加）特征的 CIN 病变的刮片中存在高甲基化水平，这可能表明 HPV 感染向癌症的快速侵袭性进程。
