Dichotomy of HIV-Sugar with Vaginal Microbes

项目概要 该提案基于 PI Mak 实验室的一项新发现，该发现表明“非静电、 特定的、可操纵的、“糖-糖”是艾滋病毒的附着因素。这种相互作用发生在 HIV 之间 受体结合之前的寡甘露糖（oligoman）和细胞 N-乙酰氨基葡萄糖（GlcNAc）。的 跨生命和非生命（病毒）领域的聚糖生物学保护导致了我们目前的提议 研究这种寡聚体-GlcNAc（或基于寡聚体）的相互作用是否可能支持某些动态- (i) 阴道微生物之间的相互作用； (二) 艾滋病毒； (iii) 它们的人类宿主。我们的研究结果表明，艾滋病毒增加一倍 使用其富含寡聚体的 HIV 包膜 (Env) 聚糖屏蔽作为附着因子可能会提供一般原则 至少部分地解释：（i）阴道乳酸菌如何提供防止艾滋病毒传播的保护； (ii) 如何 阴道淋病奈瑟菌会增加艾滋病毒传播的风险。由于许多阴道微生物都含有甘露糖- 结合蛋白和 GlcNAc 在细菌表面，我们的假设是寡聚核苷酸 N-的富集 HIV 包膜上的聚糖充当分子 Velcro 与甘露糖结合蛋白和/或 GlcNAc 糖相互作用 生殖器微生物的表面。我们进一步假设这些生殖器微生物的生物学特性 要么帮助捕获艾滋病毒以阻止其感染宿主，要么充当快递服务将艾滋病毒传递到宿主体内 主机。我们将结合使用超微结构成像技术、生物物理方法、分子病毒学 技术、飞行时间细胞计数 (CyTOF) -成像、聚糖基质辅助激光解吸电离 - 质谱成像（MALDI-MSI）和基于组织培养的上皮细胞模型系统来检查 通过基于寡聚体的结合通过两个特定目标传播艾滋病毒。在目标 1 中，我们将定义是否阴道 乳酸菌通过基于寡聚体的捕获来限制艾滋病毒的传播。在 Aim2 中，我们将评估 HIV 是否会搭便车跨越 通过寡糖介导的与淋病奈瑟菌的结合形成上皮下屏障。顺利完成本次 提案可能会产生概念证明数据，一个简单的基于寡头的机制可以解释 乳酸菌如何保护和淋病奈瑟菌如何增强艾滋病毒传播的能力分别相反。 我们还将使用 ATCC 预设的阴道微生物参考群体来评估这些基于聚糖的微生物 人口层面的互动。我们将对来自以下来源的阴道微生物组进行后续分析： 已建立且具有临床相关性的南非艾滋病毒队列。数据显示基于寡聚体的相互作用是 艾滋病毒性传播的促成因素可能为开发实用的口服药物带来新的希望 使用 GlcNAc 或甘露糖的优化版本提供自我管理的 HIV 缓解策略，考虑到这些 亲代单糖是广泛使用的、无毒的、非处方的营养保健品。

PROJECT SUMMARY This proposal is built upon a novel discovery from the PI Mak laboratory showing that a ‘non-electrostatic, specific, manipulatable, sugar-sugar’ is an attachment factor for HIV. This interaction occurs between HIV oligomannose (oligoman) and cellular N-acetylglucosamine (GlcNAc) prior to receptor engagement. The conservation of glycan biology across domains of life and the non-living (viruses) has led to our current proposal to investigate whether this oligoman-GlcNAc (or oligoman-based) interaction may underpin some of the dynamic- interplays amongst: (i) vaginal microbes; (ii) HIV; and (iii) their human host. Our findings that HIV double up to use its oligoman enriched HIV envelope (Env) glycan shield as an attachment factor may offer a general principle to account, at least in part, for: (i) how vaginal lactobacillus offers protection from HIV transmission; and (ii) how vaginal Neisseria gonorrhoeae increases the risk of HIV transmission. As many vaginal microbes have mannose- binding proteins and GlcNAc on their bacterial surface, our hypothesis is that the enrichment of oligoman N- glycans on HIV Env acts as a molecular Velcro to interact with mannose-binding proteins and/or GlcNAc sugars on the surface of genital microbes. We further hypothesize that the biology of these genital microbes would either helps trap HIV to deny their access to infect the host or acts as a courier service to deliver HIV into the host. We will use a combination of ultrastructural imaging techniques, biophysical approaches, molecular virology techniques, cytometry by time of flight (CyTOF) -imaging, glycan matrix-assisted laser desorption ionization - mass spectrometry imaging (MALDI-MSI), and tissue culture-based epithelial cell model system to examine the transmission of HIV via oligoman-based binding via two specific aims. In Aim1, we will define whether vaginal lactobacillus limits HIV transmission via oligoman-based trapping. In Aim2, we will assess if HIV hitch-hike across sub-epithelial barrier via oligoman-mediated bindings with N.gonorrhoeae. Successful completion of this proposal may result in proof of concept data that a simple oligoman-based mechanism could account for the opposing abilities of how lactobacillus protect and how N.gonorrhoeae enhance HIV transmission, respectively. We will also use pre-set vaginal microbes reference population from ATCC to evaluate these glycan-based interaction at the population level. We will perform follow-up analyses with vaginal microbiomes derived from established and clinically relevant South African HIV cohort. Data showing oligoman-based interaction is a contributing factor to sexual transmission of HIV may offer new hope for the development of a practical, orally delivery self-administrated HIV mitigation strategy using optimised versions of GlcNAc or mannose, given these parental monosaccharides are widely available, non-toxic, off-the counter, nutraceuticals.