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

随着艾滋病毒感染者预期寿命的延长，主要是由于采用了 抗逆转录病毒疗法（ART），已经变得明显，这些人差异获得广泛的 健康问题，包括口腔健康并发症，严重影响生活质量， 大量的医疗费用。我们之前发现了人类口腔癌蛋白质组谱的改变 从PLWH患者中分离的角质形成细胞，表明细胞应激升高， 提供强大的先天免疫保护。我们还证明了这些上皮细胞显示出改变的 表观遗传标记，增殖能力降低，对微生物挑战反应弱。 我们现在假设在PLWH中，口腔上皮细胞功能障碍倾向于 对继发性病毒感染的易感性，如HPV。我们召集了一个跨学科的 专家团队的专业知识包括艾滋病毒和HPV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。我们建议采用一种新的非侵入性方法收集口腔 PLWH的粘膜细胞，并进行口腔上皮的基因组和表观基因组分析（目的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HPV感染细胞扩增。 PLWH的口腔粘膜，我们已经证明表现出改变的蛋白质组，更容易受到 HPV感染与健康对照口腔上皮细胞相比（目的2）。这些研究报告将 第一个建立了HIV感染对原代上皮细胞的转录组学和表观基因组学影响， 建立一个新的基于培养的测定系统，这对未来的机制和治疗研究至关重要， 能够直接比较这些体内和体外方法， 这些特征是在PLWH中观察到的HPV易感性增加的核心。 成功完成这一R21的目标将使有针对性的基于假设的基因组或 表观基因组研究（即shRNA、CRISPR或小分子），以识别特定基因/途径 介导HPV易感性，并功能性检测HPV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治疗方法的发现和应用。 基于表观基因组学的临床干预措施;所有这些都符合NOSI NOT-DE-21-019“基本 以及与艾滋病毒/艾滋病相关的口腔健康转化研究。”

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.”