Cross Sectional Association of the Oral Microbiota and the Inflammasome with Oral HPV among HIV+ Adults

项目概要/摘要 HPV感染是最常见的性传播疾病（STI），占口咽部感染的70%。 癌的艾滋病毒感染者（PLWH）受HPV的影响不成比例，并且风险也增加 癌症的发展，即使与抑制性抗逆转录病毒疗法（ART），这表明ART可能不完全 恢复口服HPV特异性免疫。因此，了解其他可能促进 HPV在PLWH中的传播和持续存在对于癌症预防的发展至关重要， 监测策略，并减少PLWH中HPV相关恶性肿瘤的负担。口腔微生物丛 和炎性体蛋白复合物是维持免疫稳态的重要因子， 在口咽癌的发病机制中观察到。因此它们 代表了生物标志物发现和治疗的新靶点。我们的长期目标是研究这些 一些因素可能导致免疫失调，并可能影响口腔HPV感染的自然史。 PLWH，以建立一套HPV相关恶性肿瘤的早期检测标志物， 治疗干预。我们假设口腔微生物组的生态失调和炎性小体的失调 可能是HPV相关并发症的危险因素，并可能有助于抑制免疫系统 促进PLWH中随后的炎症。这项横断面研究计划招募200名病毒学 来自波多黎各社区组织的受抑制PLWH（≥21岁且<49岁的男性和女性） (PR)CONCRA，提供专门治疗艾滋病毒和性传播疾病的预防和保健服务。PLWH 参加PR CONCRA的人构成了一个独特的高危人群，其中艾滋病毒和口腔HPV的流行率 感染率更高，使我们的专家团队在艾滋病毒，HPV，流行病学，微生物学，口腔健康和 生物信息学可以高效地建立这些关系。这项建议的具体目标是：（1） 表征唾液中的口腔微生物群，并研究其与HPV状态和HPV基因型的关系， 和（2）确定炎性体蛋白（NLRP 3、AIM 2和半胱天冬酶）的水平之间的关联。 1、IL-1β和裂解的IL-1β）和炎症标志物（IL-1β、IL-6、IL-8、TNF-α、IFN-γ）与口腔HPV感染的关系 和HPV基因型在收集唾液样本后，参与者将接受口腔健康评估。 将通过问卷调查收集社会人口统计学、行为、临床特征和风险因素。 受试者将接受稳定的抑制性ART至少6个月，以尽量减少由于以下原因引起的炎症信号： 艾滋病毒活跃复制。我们将通过对16 S rDNA基因和HPV进行测序来表征口腔微生物群。 基因分型。炎性体蛋白和炎性标志物将通过蛋白质印迹法测量， 多重ELISA检测。

PROJECT SUMMARY/ABSTRACT HPV-infection is the most common sexually transmitted disease (STI), and it accounts for 70% of oropharyngeal cancers. People living with HIV (PLWH) are disproportionately affected by HPV and are also at an increased risk of cancer development, even with suppressive antiretroviral therapy (ART), suggesting that ART may not fully recover oral HPV-specific immunity. Therefore, understanding other underlying factors that could facilitate the transmission and persistence of HPV in PLWH is crucial for the development of cancer prevention and surveillance strategies, and reduce the burden of HPV-associated malignancies in PLWH. The oral microbiota and inflammasome protein complex are essential factors for maintaining immune homeostasis factors, which are altered by HIV infection and have been observed in the pathogenesis of oropharyngeal cancer. Thus they represent novel targets for biomarker discovery and therapeutics. Our long-term goal is to investigate how these factors could contribute to immune dysregulation, and may affect the natural history of oral HPV infection in PLWH, in order to establish a set of early detection markers for HPV-related malignancies and personalized treatment interventions. We hypothesize that dysbiosis of the oral microbiome and inflammasome dysregulation can be risk factors for HPV-related complications and can contribute to the dampening the immune system promoting subsequent inflammation in PLWH. This cross-sectional study plans to recruit 200 virologically suppressed PLWH (men and women ≥21 and <49 years old) from a community-based organization, Puerto Rico (PR) CONCRA, that offers preventive and health services specialized in the treatment of the HIV and STI. PLWH who attends PR CONCRA constitute a unique high-risk group where the prevalence of both HIV and oral HPV infection is higher, allowing our team of experts in HIV, HPV, epidemiology, microbiology, oral health and bioinformatics to establish these relationships with high efficiency. The specific aims of this proposal are: (1) to characterize the oral microbiota in saliva and investigate its relationship with HPV status and HPV genotypes, and (2) to determine the association between the levels of inflammasome proteins (NLRP3, AIM2, and caspase- 1, IL-1β and cleaved IL-1β) and inflammation markers (IL-1β, IL-6, IL-8, TNF-α, IFN-γ) with oral HPV infection and HPV genotypes. After collection of saliva samples, participants will undergo oral health evaluations. Sociodemographic, behavioral, clinical characteristics, and risk factors will be collected through a questionnaire. Participants will be on stable, suppressive ART for at least 6 months to minimize inflammatory signals due to HIV active replication. We will characterize the oral microbiota by sequencing the 16S rDNA gene and HPV genotypes by PCR. Inflammasome proteins and inflammatory markers will be measured by Western Blot and multiplex ELISA respectively.