Investigations Into The Molecular Pathogenesis Of Cervical Glandular Neoplasias

摘要 每年有超过12，000名妇女被诊断患有宫颈癌，这是一种HPV相关的恶性肿瘤， 宫颈癌有两种主要的组织学形式：鳞状细胞癌（SCC）和 腺癌（ADENO）。重要的是，SCC可以通过宫颈细胞学筛查来预防 而ADENO不能。虽然有针对HPV的预防性疫苗，但HPV的摄取 美国的疫苗接种一直很差，已经有几代女性接触过疫苗。 HPV感染，并不会帮助HPV疫苗接种。因此，子宫颈普查的费用超过6美元， 至少在几十年内，每年需要10亿美元。了解脑出血的分子发病机制 ADENO具有重要的公共卫生价值，以促进ADENO的预防，目前， 筛查策略不充分。在美国和欧洲，HPV 16、18和45感染占>95%。 AIS/ADENO我们检测了HPV 16/18/45 AIS（n=27）和宫颈上皮内瘤样病变的病毒甲基化， 肿瘤3级（CIN 3）（n=63）病例与90例HPV 16/18/45对照感染<CIN 2相比，发现 HPV 16的AUC分别为0.99和0.82。此外，我们有初步数据显示， AIS病变中宿主基因组的甲基化水平显著高于对照组（p<0.001 （<CIN2）。此外，我们还采用了HPV-DNA捕获NGS检测法来前瞻性地评估病毒， 整合，并发现了在整个筛选过程中保持的特异性HPV 18病毒整合。我们 进一步发现该整合位点具有HPV整合读数的逐渐增加，导致， 在组织学鉴定偶发CIN 3病变之前，覆盖率有一个显著的峰值。 NGS检测到的整合可能代表了一个扩大的恶性细胞克隆，并显示出强大的潜力 用于临床应用。 我们建议调查HPV和ADENO前体的自然史（即，样品之前， 诊断ADENO和AIS），目的是定义ADENO的分子前体， 立即治疗，从而防止ADENO。我们将利用大量的 HPV 16、18和45阳性女性（n > 20，000），绝大多数ADENO发生在其中，以衡量宿主 以及HPV病毒甲基化变化和HPV病毒整合来从分子上定义HPV的自然史。 阿德诺。我们将比较和对比这些发现与更好地了解自然史的SCC。最后， 我们将使用HPV基因型、HPV病毒和宿主， 基因表观遗传学和HPV病毒整合。

ABSTRACT More than 12,000 women are diagnosed with cervical cancer, an HPV-associated malignancy, annually in the U.S. There are two major histologic forms of cervix cancer: squamous cell carcinoma (SCC) and adenocarcinoma (ADENO). Importantly, SCC can be prevented through cervical cytology screening programs, whereas ADENO cannot. While there is a prophylactic vaccine against HPV, uptake of HPV vaccination in the U.S. has been poor and there are several generations of women who are already exposed to HPV infection and will not be helped by HPV vaccination. Therefore, cervical screening at a cost of over $6 billion per year will be needed for several decades at least. Understanding the molecular pathogenesis of ADENO has important public health value in order to facilitate the prevention of ADENO, for which current screening strategies are inadequate. In the US and Europe, HPV16, 18, and 45 infections account for >95% of AIS/ADENO. We examined viral methylation of HPV16/18/45 AIS (n=27) and cervical intraepithelial neoplasia grade 3 (CIN3) (n=63) cases compared to 90 HPV16/18/45 control infections with <CIN2, and found AUC's of 0.99 and 0.82 for HPV16, respectively. In addition, we have preliminary data showing that methylation levels of a host gene panel is significantly higher in AIS lesions (p<0.001) compared to controls (<CIN2). Furthermore, we have employed an HPV-DNA capture NGS assay to prospectively evaluate viral integration and found a specific HPV18 viral integration that is maintained for the entirety of the screening. We further found that this integration site has a progressive increase in HPV-integration reads leading up to, and with a dramatic spike in coverage directly preceding, the histological identification of an incident CIN3 lesion. The NGS detected integration likely represents an expanding malignant cell clone and shows strong potential for clinical application. We propose to investigate the natural history of HPV and ADENO precursors (i.e., samples prior to the diagnosis of ADENO and from AIS) with the goal of defining molecular precursors of ADENO, which could be amenable to immediate treatment and thus prevent ADENO. We will leverage cohorts with large numbers of HPV16, 18, and 45-positive women (n > 20,000) in whom the vast majority of ADENO occur, to measure host and HPV viral methylation changes and HPV viral integration to molecularly define the natural history of ADENO. We will compare and contrast these findings with the better known natural history of SCC. Ultimately, we will develop a risk-prediction model for AIS/ADENO precursors using HPV genotypes, HPV viral and host gene epigenetics, and HPV viral integration.