Next Generation of HPV and Cervical Cancer Research in HIV+ Women

艾滋病毒（+）妇女有很高的风险，宫颈癌前病变和癌症，以及感染人类乳头瘤病毒 (HPV)宫颈癌/癌前病变的病毒原因。遗传/表观遗传方法的最新进展提供了 以前无法实现的机会，研究HPV病毒因素和其他局部感染对 HPV的自然史和HIV（+）妇女宫颈疾病的发展。根据这项建议，我们将使用 下一代（下一代）测序进行精确的HPV基因组分析，能够确定 在两个或更多个时间点检测到的给定HPV类型是否是相同的确切病毒感染， 同一类型的HPV。这些数据将用于全面研究：HPV的类型特异性差异 持续性及其与癌前病变的关系; HPV再激活的发生率和再激活的频率 HPV持续存在并导致癌前病变;免疫状态对HPV自然感染中每个步骤的影响 历史如果宫颈HPV可以重新激活并发展为癌前病变， 65岁（在一般人群中进行）。此外，HPV DNA甲基化将被研究，因为我们 发现HPV L1/L2区域的甲基化与癌前病变/癌症密切相关， 这表明在HIV（+）妇女中甲基化和癌前病变之间可能存在类似的联系。本研究 因此，将提供对与宫颈疾病发展相关的表观遗传修饰的深入了解， 可用于提高HPV检测的特异性和阳性预测值。重要的是， 宫颈阴道微生物组可能影响HPV自然史和HPV DNA甲基化， 也将被研究。我们小组的一项初步研究显示，HPV患病率降低，相对丰度较高 卷曲乳杆菌而不是其他乳杆菌属物种，并过渡到L.卷曲群落状态 型与HPV感染率降低有关。洛杉矶crispatus的结果特别有希望，因为 即使在低CD 4的情况下也观察到保护作用。然而，微生物群和HPV的关系需要 在任何未来的益生菌之前，在适当设计的足够规模的HIV（+）队列研究中进行研究 可以考虑进行干预研究。目前关于微生物群的影响的数据很少。 HPV自然史/进展（基于目标1）。我们还将研究微生物群和HPV甲基化 （建立在目标2上），因为局部微生物群显示出改变邻近组织中的甲基化。总的来说，这些 综合研究解决了HPV自然史/进展的关键方面， 癌症预防。这项研究将利用从WIHS每半年收集的标本，WIHS是世界上最大的，最长的 HIV（+）（N=2793）和高危HIV（-）（N=975）女性的足月队列。具体目标是，在艾滋病毒（+） 研究：（i）使用下一代HPV疫苗， “精确”HPV基因组检测;（ii）HPV DNA甲基化及其与癌前风险的关系; 微生物组对HPV自然史、HPV甲基化和HPV进展的影响。

HIV(+) women have high risk of cervical precancer and cancer as well as infection with human papillomavirus (HPV), the viral cause of cervical precancer/cancer. Recent advances in genetic/epigenetic methods provide previously unachievable opportunities to study the HPV viral factors and other local infectious influences on the natural history of HPV and development of cervical disease in HIV(+) women. Under this proposal, we will use next generation (next-gen) sequencing to conduct precision HPV genomic analysis able to determine whether a given HPV type detected at two or more time points is the same exact viral infection versus different HPVs of the same type. These data will be used to comprehensively study: type-specific differences in HPV persistence and their relation with precancer; the occurrence of HPV reactivation and how often reactivated HPV persists and leads to precancer; the impact of immune status on each of these steps in HPV natural history. If cervical HPV can reactivate and progress to precancer, it would preclude screening cessation at age 65 years (which is done in the general population). Furthermore, HPV DNA methylation will be studied, as we found methylation in the HPV L1/L2 region very strongly associated with precancer/cancer, and pilot data suggest similar associations between methylation and precancer may exist for HIV(+) women. This research therefore will provide insight into the epigenetic modifications related to the development of cervical disease, and could possibly be used to improve the specificity and positive predictive value of HPV testing. Importantly, the cervicovaginal microbiome may influence both the HPV natural history and HPV DNA methylation and will also be studied. A pilot study by our group showed reduced HPV prevalence with high relative abundance of Lactobacillus crispatus but not other Lacobacillus species, and transition to a L. crispatus community state type was associated with reduced incident HPV. The L. crispatus results were especially promising since the protective effects were observed even with low CD4. However, the microbiota and HPV relationship needs to be studied in appropriately designed HIV(+) cohort studies of adequate size before any future probiotic intervention studies may be considered. There are currently little data regarding the impact of the microbiota on HPV natural history/progression (building on Aim 1). We will also study the microbiota and HPV methylation (building on Aim 2), as local microbiota was shown to alter methylation in neighboring tissue. Overall, these integrated studies address critical aspects of HPV natural history/progression, with significant implications to cancer prevention. This study will utilize semiannually collected specimens from the WIHS, the largest, long term cohort of HIV(+) (N=2793) and high risk HIV(-) (N=975) women. The Specific Aims are, in HIV(+) women, to study: (i) The role of reactivation in HPV natural history and precancer risk, using next-gen “precision” HPV genomic assays; (ii) HPV DNA methylation and its relation with precancer risk; The microbiome's impact on HPV natural history, HPV methylation, and HPV progression.