Impact of HIV, oral microbiome and mycobiome on oral HPV persistence

摘要 口腔人乳头瘤病毒（HPV）感染和HPV相关的乳头瘤和癌症继续上升， 尤其是艾滋病毒感染者（PLWH）。特别是，HPV相关的口腔和口咽 癌症正以惊人的速度增长。通过大量基于人群的纵向研究，我们的工作 表明口腔病变和头颈癌的风险增加，不仅HPV 16， 皮肤的β和γ型HPV。通过研究风险因素，我们发现了吸烟与 和艾滋病毒血清状态，口腔高危α和β-HPV检测的流行率增加。除了 口腔HPV，HIV影响口腔微生物组的组成，这也与口腔HPV感染增加有关。 对口腔HPV感染的易感性、持续性和进展为HPV驱动的癌症。综上所述各项 研究结果表明，一个持续的公共卫生关注HPV相关的口腔疾病的PLWH。拟议 研究将利用从N=2046名男性和女性PLWH和N=1334名PLWH中收集的重复口腔唾液样本 入组MACS-WIHS联合队列研究（MWCSS）的HIV[-]个体。具体来说，我们将 调查口腔微生物组和真菌组与获得风险之间的关联， 高危α、β和γ口腔HPV的持续存在。这项研究的科学前提是， 由HIV和吸烟引起的口腔微生物组/真菌组的改变影响口腔HPV自然 历史我们将使用我们团队开发的新型测序方法，包括下一代测序 和宏基因组学，以分析HPV病毒组（α、β和γ HPV）、口腔细菌（16 S rRNA）和 真菌群落（使用最近开发的ITS 1测定，其对念珠菌的敏感性增加， Burk实验室）、功能基因能力（由鸟枪法宏基因组测序确定）和细胞因子 分布图（来自口腔唾液蛋白分析）。口腔微生物组/真菌组之间的差异 将评估艾滋病毒感染者和艾滋病毒[-]个人，以及艾滋病毒疾病和吸烟的标志物， 口腔HPV持久性。利用在MWCCS参与者中收集的系列样本，我们将 表征口腔微生物组/真菌组随时间的变化对CD 4计数和HIV RNA水平的响应， 吸烟和口腔微生物组/真菌组，以解决以下具体目标：1）确定 HIV和吸烟对口腔HPV持续性风险的联合影响; 2）研究 口腔微生物组/真菌组和口腔HPV持续存在的风险; 3）表征微生物功能和口腔HPV持续存在的风险。 与口腔HPV持续性风险相关的炎症/免疫途径。我们假设口腔 受艾滋病毒和吸烟影响的微生物组和真菌组表现出明显更高的毒力比例， 这些因素反过来又会使口腔屏障免疫和完整性失调，促进口腔高危疾病的持续存在。 HPV类型与口腔癌和口咽癌风险增加相关。

ABSTRACT Oral human papillomavirus (HPV) infections and HPV-associated papillomas and cancers continue to rise, especially in people living with HIV (PLWH). Specifically, rates of HPV-associated oral and oropharyngeal cancer are increasing at an alarming rate. Using large population-based longitudinal studies, our work indicated an increased risk of oral lesions and head and neck cancers with not only HPV16, but unexpectedly cutaneous beta and gamma HPVs as well. Studying risk factors, we identified an interaction between smoking and HIV serostatus with increased prevalence of oral high-risk alpha and beta-HPV detection. In addition to oral HPV, HIV affects the composition of the oral microbiome, which has also been associated with increased susceptibility to oral HPV infection, persistence, and progression to HPV-driven cancer. Taken together, these findings indicate a continuing public health concern of HPV-related oral diseases for PLWH. The proposed study will leverage repeat oral saliva samples collected from N=2046 male and female PLWH, and N=1334 HIV[-] individuals enrolled in the MACS-WIHS Combined Cohort Study (MWCSS). Specifically, we will investigate the associations between the oral microbiome and mycobiome and risk of acquisition and persistence of high-risk alpha, beta, and gamma oral HPV. The scientific premise of the study is that alterations in the oral microbiome / mycobiome resulting from HIV and smoking influence oral HPV natural history. We will use novel sequencing methods developed by our team, including next generation sequencing and metagenomics, to profile the HPV virome (alpha, beta, and gamma HPV), oral bacterial (16S rRNA) and fungal communities (using a recently developed ITS1 assay with increased sensitivity for Candida developed in the Burk lab), functional gene capacity (determined from shotgun metagenomic sequencing), and cytokine profiles (derived from oral saliva protein analyses). Differences in the oral microbiome / mycobiome between PLWH and HIV[-] individuals will be assessed, as well as markers of HIV disease and smoking, with respect to oral HPV persistence. Taking advantage of serial samples collected in MWCCS participants, we will characterize oral microbiome / mycobiome changes over time in response to CD4 count and HIV RNA level, smoking, and the oral microbiome / mycobiome to address the following specific aims: 1) Determine the combined effects of HIV and smoking on risk of oral HPV persistence; 2) Study the associations between the oral microbiome / mycobiome and risk of oral HPV persistence; 3) Characterize the microbial function and oral inflammation/immune pathways associated with risk of oral HPV persistence. We hypothesize that the oral microbiome and mycobiome influenced by HIV and smoking exhibit significantly higher proportions of virulence factors, which in turn dysregulate the oral barrier immunity and integrity, facilitating persistence of oral high-risk HPV types associated with increased risks for oral and oropharyngeal cancer.