Short Chain Fatty Acids: Route for Facilitation of Oral HPV via Inflammasome Dysregulation in People Living with HIV

项目总结/摘要 口腔微生物群和炎性体蛋白复合物是维持免疫的重要因素 体内平衡，这是改变了艾滋病毒感染，并已观察到的发病机制， 口咽癌口腔中的短链脂肪酸（SCFA），如丁酸盐， 牙周组织损伤，它可以作为一种致癌代谢物，并导致HPV相关的发展， 口腔和咽部致癌。此外，已发现丁酸盐调节以下转录： 免疫相关基因特别是，它上调局部炎症，同时下调 干扰素相关基因和上调炎性小体相关标志物，先天免疫所必需的 对病毒感染的反应。因此，丁酸可以通过以下方式在免疫功能障碍中发挥重要作用： 炎症体失调，促进口腔HPV感染，增加患有HPV的人患癌症的风险。 艾滋病毒（艾滋病毒携带者）。因此，我们的总体目标是量化PLWH与非HPV感染的口腔SCFA 中心假设是，患有口腔HPV感染的PLWH将具有更高水平的丁酸盐，这将是 与牙周病，口腔微生物群落的生态失调，更多的炎症和更多的 炎性小体调节障碍。本补充计划使用来自50个病毒学抑制的PLWH的样本 (25 HPV+和25 HPV-），年龄≥21岁且<49岁。本提案的具体目标 （i）量化PLWH唾液中微生物SCFA的水平，并评估其与感染的关系， HPV基因型（高风险vs.低风险）和（ii）确定SCFA水平与 牙周病、口腔微生物菌群失调、炎性体失调和炎症水平 标记。SCFA将通过液相色谱法和质谱法进行测量。我们将利用 HPV基因分型（PCR）、口腔微生物组数据（16 s rDNA分析）、炎性小体蛋白水平（蛋白质印迹） 和来自母研究的炎症标志物（多重ELISA）。该奖项将直接支持Yabdiel先生 Ramos-Valerio在微生物组研究和数据分析方面拥有专业知识，包括大规模的 宏基因组学数据和统计分析以及流行病学研究，重点是艾滋病毒、HPV和癌症。他 还将参加课程和重点指导，以扩大他在设计， 实施并完成临床/转化研究。最后，他将进一步发展学术 技能，这将确保他的成功，独立的研究生涯，包括负责任的研究行为， 科学写作，口头沟通技巧，语法，跨学科合作研究。这 补充将为他提供资源，以实现他的目标，被接受在MD/博士课程 并最终成为一名专门研究病原体驱动的癌症的独立研究医生。

PROJECT SUMMARY/ABSTRACT The oral microbiota and inflammasome protein complex are essential factors for maintaining immune homeostasis, which are altered by an HIV infection and have been observed in the pathogenesis of oropharyngeal cancer. Short chain fatty acids (SCFA) in the oral cavity, such as butyrate, exacerbates periodontal tissue damage, which can act as an oncometabolite and lead to the development of HPV-related oral and pharyngeal carcinogenesis. Furthermore, butyrate has been found to regulate the transcription of immune-related genes. Particularly, it upregulates local inflammation while downregulates the expression of interferon-related genes and upregulates inflammasome-related markers, necessary for the innate immune response to a viral infection. Therefore, butyrate could play an essential role on immune dysfunction by dysregulating the inflammasome and facilitating oral HPV infection, increasing cancer risk in people living with HIV (PLWH). Therefore, our overall objective is to quantify oral SCFA in PLWH with an without HPV infection with the central hypothesis that PLWH with oral HPV infection will have higher levels of butyrate, which will be associated with periodontal disease, dysbiosis of the oral microbiome, more inflammation and more inflammasome dysregulation. This supplement plans to use samples from 50 virologically suppressed PLWH (25 HPV+ and 25 HPV-), aged ≥21 and<49 years old from the parent study. The specific aims of this proposal is (i) to quantify the levels of microbial SCFA in saliva of PLWH and assess their relationship with infection and HPV genotypes (high-risk vs. low-risk) and (ii) To determine the association between the levels of SCFA with periodontal disease, oral microbiome dysbiosis, inflammasome dysregulation and levels of inflammation markers. SCFA will be measured by liquid chromatography followed by mass spectrometry. We will leverage HPV genotyping (PCR), oral microbiome data (16s rDNA profiling), inflammasome proteins levels (western blot) and inflammatory markers (multiplex ELISA) from the parent study. This award will directly support Mr. Yabdiel Ramos-Valerio in achieving expertise in microbiome research and data analysis, including large-scale metagenomics data and statistical analysis and epidemiological research focused on HIV, HPV, and cancer. He will also take courses and focused mentorship on expanding his knowledge and experience in designing, implementing, and completing a clinical/translational research study. Finally, he will further develop the academic skills that will ensure his successful, independent research career, including responsible conduct of research, scientific writing, oral communication skills, grantsmanship, and collaborative interdisciplinary research. This supplement will provide him with resources toward achieving his goal of being accepted in MD/PhD programs and eventually become an independent research physician specializing in pathogen-driven cancers.