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

项目摘要/摘要 人乳头瘤病毒感染是最常见的性传播疾病，占口咽疾病的70% 癌症。艾滋病毒携带者(PLWH)感染HPV的比例更高，风险也更高 即使使用抑制性抗逆转录病毒疗法(ART)，癌症的发展也是如此，这表明ART可能并不完全 恢复口服HPV特异性免疫。因此，了解其他潜在因素可能有助于 HPV在PLWH中的传播和持续对于癌症预防和治疗的发展至关重要 监测策略，并减少PLWH中HPV相关恶性肿瘤的负担。口腔微生物区系 和炎症体蛋白复合体是维持免疫动态平衡的必要因素，这些因素包括 被HIV感染改变，并在口咽癌的发病机制中观察到。因此，他们 代表了生物标记物发现和治疗的新靶点。我们的长期目标是调查这些 因素可能导致免疫失调，并可能影响口腔HPV感染的自然病史。 PLWH，以便建立一套针对HPV相关恶性肿瘤的早期检测标记物，并进行个体化 治疗干预。我们假设口腔微生物群失调和炎症体失调 可能是HPV相关并发症的危险因素，并可能导致免疫系统受抑 促进PLWH的后续炎症。这项横断面研究计划招募200名病毒学研究人员 来自波多黎各社区组织的Pwh(21岁和49岁的男性和女性≥)被压制 (PR)Concra，提供专门治疗艾滋病毒和性传播感染的预防和保健服务。PLWH 世卫组织参加PR Concra构成了一个独特的高危群体，在那里HIV和口腔HPV的流行 感染率更高，使我们的专家团队能够在艾滋病毒、HPV、流行病学、微生物学、口腔健康和 生物信息学来高效地建立这些关系。这项建议的具体目的是：(1) 口腔唾液微生物区系特征及其与HPV感染状态和HPV基因分型的关系 以及(2)确定炎症体蛋白(NLRP3、AIM2和caspase-1)水平之间的关系。 IL-1β和裂解IL-1β)和炎症标志物(IL-1β、IL-6、IL-8、肿瘤坏死因子-α、干扰素-γ)与口腔HPV感染的关系 和HPV基因分型。在采集唾液样本后，参与者将接受口腔健康评估。 将通过问卷收集社会人口学、行为、临床特征和危险因素。 参与者将接受至少6个月的稳定、抑制性ART，以最大限度地减少因以下原因而产生的炎症信号 艾滋病毒主动复制。我们将通过对16S rDNA基因和HPV进行测序来表征口腔微生物区系 用聚合酶链式反应进行基因分型。炎症体蛋白和炎症标志物将通过Western Blot和 分别采用多重酶联免疫吸附试验。

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.