Epigenetics of dysfunctional oral epithelium in people living with HIV and risk for HPV infection

随着艾滋病毒携带者(PLWH)预期寿命的增加，主要是由于引入了 抗逆转录病毒疗法(ART)，很明显，这些个体获得的差异很大。 健康问题，包括严重影响生活质量和引起的口腔健康并发症 高额的医疗费用。我们之前发现了一种改变的人类口腔蛋白质组图谱 从PLWH患者中分离的角质形成细胞，表明细胞应力增加和能力降低 提供强大的先天免疫保护。我们还证明了这些上皮细胞表现出改变 表观遗传标记，增殖能力降低，对微生物挑战的反应较弱。 我们现在假设，在PLWH中，口腔上皮细胞功能障碍容易发生 对继发性病毒感染的易感性，如HPV。我们已经组建了一个跨学科的 专家团队的专长包括艾滋病毒和人乳头状瘤病毒病毒学，口腔和上皮细胞生物学， 表观基因组学和生物信息学。我们建议应用一种新的非侵入性口腔采集方法 并对口腔上皮进行基因组和表观基因组分析(目标1)。 此外，使用相关的HPV感染模型，我们希望确定这些细胞是否从 我们已经证实的PLWH的口腔粘膜呈现出一种改变的蛋白质组，更容易受到 HPV感染与来自健康对照组的口腔上皮细胞相比(目标2)。这些研究将是 第一次建立了艾滋病毒感染对原代上皮细胞的转录和表观基因组效应， 建立一种新的基于培养的检测系统，对未来的机制和治疗研究至关重要，以及 能够在体内和体外方法之间进行直接比较，以强有力地识别关键分子 PLWH中HPV易感性增加的核心特征。 成功完成R21的目标将使基于假设的有针对性的基因组或 表观基因组学研究(即shRNA、CRISPR或小分子)以确定特定的基因/途径 介导人乳头瘤病毒的易感性，并从功能上检测人乳头瘤病毒感染水平。验证表观遗传学基础 PLWH对HPV感染易感性的研究将最终指导新的发现和应用 基于表观基因组学的临床干预；全部与NOSI NOT-DE-21-019“BASIC”的目标一致 以及与艾滋病毒/艾滋病有关的口腔健康翻译研究。

With the increase in life expectancy of people living with HIV (PLWH), principally due to the introduction of antiretroviral therapy (ART), it has become evident that these individuals differentially acquire a wide range of health problems, including oral health complications, which severely impact quality of life and incur substantial healthcare costs. We previously identified an altered proteomic profile of human oral keratinocytes isolated from PLWH patients, suggesting elevated cellular stress and reduced ability to provide robust innate immune protection. We also demonstrated that these epithelial cells display altered epigenetic markers, reduced proliferative capacity and respond weakly to microbial challenges. We now hypothesize that in PLWH, oral epithelial cell dysfunction predisposes towards susceptibility to secondary viral infections, such as HPV. We have assembled an interdisciplinary team of experts whose expertise encompasses HIV and HPV virology, oral and epithelial cell biology, epigenomics and bioinformatics. We propose to apply a novel non-invasive method of collecting oral mucosal cells from PLWH and conduct genomic and epigenomic analyses of the oral epithelium (Aim 1). Additionally, using a relevant HPV infection model, we wish to determine if such cells expanded from the oral mucosa of PLWH, which we have demonstrated exhibit an altered proteome, are more susceptible to HPV infection when compared to oral epithelial cells from healthy controls (Aim 2). These studies will be the first to establish the transcriptomic and epigenomic effects of HIV infection on primary epithelial cells, establish a new culture-based assay system so critical for future mechanistic and therapeutic studies, and enable direct comparisons between these in vivo and in vitro methods to robustly identify key molecular features that are central to the increased HPV susceptibility seen in PLWH. Successful completion of the goals of this R21 will enable targeted hypothesis-based genomic or epigenomic studies (i.e. shRNAs, CRISPR, or small molecules) to identify specific genes/pathways mediating the HPV susceptibility, and functionally test HPV infection levels. Validating the epigenetic basis of susceptibility to HPV infection in PLWH will eventually guide the discovery and application of novel epigenomic-based clinical interventions; all consistent with the goals of the NOSI NOT-DE-21-019 “Basic and translational oral health research related to HIV/AIDS.”