PO.MCB06.02 · 分子与细胞生物学

甲基化DNA标志物在准确检测高危HPV阳性宫颈病变中的潜力

The potential of methylated DNA markers for accurate detection of high-risk, HPV-positive cervical lesions

编号 1959 展板 11 时间 4/20 09:00–12:00 区域 Section 22 主讲 Roshni Saravanan, BS;MS;PhD

分会场 DNA Methylation

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作者与单位 Authors & Affiliations

Roshni Saravanan¹, Mary Jo Fackler¹, Madison Pleas¹, Wenfei Xia¹, Tomisin Adebari¹, Gang Yu¹, Liqun Zhang¹, Suzette Jordaan², Eunice Van Den Berg², Pamela Michelow², Reubina Wadee², Maureen Joffe³, Wenlong Carl Chen³, Youxiang Wang⁴, Leslie Cope¹, Saraswati Sukumar¹

¹Department of Oncology, Johns Hopkins University School of Medicine, Baltimore, MD,²Department of Anatomical Pathology, University of the Witwatersrand/National Health Laboratory Service, Johannesburg, South Africa,³Strengthening Oncology Services Research Unit, Faculty of Health Sciences, University of the Witwatersrand, Johannesburg, South Africa,⁴Atila Biosciences, Sunnyvale, CA

摘要 Abstract

中文摘要

目的：宫颈涂片细胞学在检测宫颈病变方面表现出高度变异性和有限的敏感性。虽然WHO推荐的HPV检测具有高敏感性，但其特异性有限。我们的目标是确定DNA甲基化标志物能否同时提高敏感性和特异性，从而实现比细胞学和HPV检测更有效的阴道镜和活检分诊。实验设计：我们此前发表了一项5基因(TBXT、MOS、FMN、EDNRB和ZNF671)DNA甲基化检测方法，在细胞学和组织学样本中检测宫颈癌具有高敏感性和特异性[1]。在本研究中，我们检测了222份宫颈刷取样本(n = 222)，这些样本具有已知的细胞学结果但无可用的组织学结果，保存于CytoLyt中，由南非约翰内斯堡的国家卫生服务筛查诊所选取并去标识化。样本采用Atila Biosystems进行高危HPV变异检测，并采用定量多重甲基化特异性PCR(QM-MSP)进行累积5基因甲基化检测。统计学检验包括基因特异性阈值的约登指数、单个基因的比值比评分、Mann-Whitney检验以及受试者工作特征曲线下面积(ROC AUC);P值≤0.05被认为具有显著性。结果：在222份宫颈刷取样本中，HPV在HSIL(87/95,92%)和LSIL(47/57,82%)中的流行率相近，在约半数的NIEL(39/70,56%)中检出。因此，HPV对HSIL的检测高度敏感，但对低危LSIL或NIEL的特异性很差。相比之下，在HPV阳性样本中，为区分高危HSIL与低危LSIL/NEIL，QM-MSP检测显示出高水平的敏感性:78.16%[CI 68.39 - 85.55],此外还具有高水平的特异性:95.35%[CI 88.64 - 98.18],ROC AUC = 0.890[CI 83.71 - 94.37]。HSIL与LSIL/NIEL之间的甲基化水平存在高度显著(P<0.0001)的差异。在少量HPV阴性样本中，5基因组合的高甲基化见于5/8(63%)HSIL、0/10(0%)LSIL和1/31(3%)NIEL,提示QM-MSP可能独立于HPV状态。结论：这些发现提示,5基因组合检测出很大比例的HSIL,而大多数LSIL和NIEL保持阴性。因此,该检测可作为HPV阳性病例的有效分诊工具,加快HSIL女性的诊治,同时可能将LSIL和NIEL的阴道镜转诊减少多达95%。这可减少低危病变反复活检操作相关的成本和风险。参考文献:[1] DOI: 10.1186/s13148-024-01669-z

查看英文原文 English abstract

Purpose: Cervical smear cytology exhibits high variability and limited sensitivity in detecting cervical lesions. While the HPV test, recommended by WHO, offers high sensitivity, it has limited specificity. Our goal was to determine whether DNA methylation markers could improve both sensitivity and specificity, thereby enabling more effective triage to colposcopy and biopsy than cytology and HPV testing. Experimental Design: We previously published a 5-gene ( TBXT, MOS, FMN, EDNRB, and ZNF671 ) DNA methylation assay with high sensitivity and specificity for detecting cervical cancer in cytology and histology samples [1]. In the current study, we tested cervical brush samples (n = 222) with known cytology but no available histology that were preserved in CytoLyt, selected and deidentified at the National Health Service screening clinic in Johannesburg, South Africa. Samples were tested for both high-risk HPV variants (Atila Biosystems) and for cumulative 5-gene methylation utilizing Quantitative Multiplex Methylation-Specific PCR (QM-MSP). Statistical tests included the Youden Index for gene-specific thresholds, odds ratio scores for individual genes, Mann-Whitney test, and Receiver Operating Characteristic Area Under the Curve (ROC AUC); a P value ≤ 0.05 was considered significant. Results: Among the 222 cervical brush samples, HPV was prevalent in similar numbers of HSIL (87/95, 92%) and LSIL (47/57, 82%), and in about half of the NIEL (39/70, 56%). Thus, HPV was highly sensitive for the detection of HSIL, but very poorly specific for low-risk LSIL or NIEL. In contrast, among HPV+ samples, to distinguish between high risk HSIL and low risk LSIL/NEIL, the QM-MSP assay showed a high level of sensitivity: 78.16% [CI 68.39 - 85.55] and, in addition, a high level of specificity: 95.35% [CI 88.64 - 98.18], and ROC AUC = 0.890 [CI 83.71 - 94.37]. A highly significant (P<0.0001) difference in methylation levels was observed between HSIL and LSIL/NIELs. In the small set of HPV-negative samples, hypermethylation of the 5-gene panel was observed in 5/8 (63%) HSIL, 0/10 (0%) LSIL, and 1/31 (3%) NIEL, suggesting that QM-MSP may be independent of the HPV status. Conclusions: These findings suggest that the 5-gene panel detected a large proportion of HSIL, while the majority of LSIL and NIEL remained negative. Thus, this assay could serve as an effective triage tool for HPV+ cases, expediting care for women with HSIL while potentially reducing referrals for colposcopy in LSIL and NIEL by up to 95%. This could reduce costs and risks associated with repeated biopsy procedures for low-risk lesions. Reference: [1] DOI: 10.1186/s13148-024-01669-z

利益披露 Disclosure

R. Saravanan, None.. M. Fackler, None.. M. Pleas, None.. W. Xia, None.. T. Adebari, None.. G. Yu, None.. L. Zhang, None.. S. Jordaan, None.. E. Berg, None.. P. Michelow, None.. R. Wadee, None.. M. Joffe, None.. W. Chen, None.. Y. Wang, None.. L. Cope, None.. S. Sukumar, None.

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