Integrating microbial glycan arrays with genomic sequences to study host microbe interactions

摘要：虽然成功的免疫依赖于宿主免疫因子快速识别和 对各种微生物决定因素的反应，微生物多聚糖往往是第一个也是最重要的 与宿主免疫因子的接触点。然而，大多数天然免疫和获得性免疫的结合特异性 影响微生物多聚糖的因素在很大程度上仍不清楚。此外，尽管几乎普遍使用了 基因组测序方法表征微生物区系，这些方法在很大程度上无法识别微生物 基于其独特的多糖特征，排除了与相关的不同微生物菌株的鉴定 当使用这种方法时，糖链结构。为了有效地克服目前研究中的局限性 宿主-微生物相互作用，必须开发一个可供各种研究人员使用的平台 定义宿主如何与微生物多聚糖相互作用。我们的长期目标是开发一个 可用于定义先天免疫和获得性免疫网络的微生物多糖和基因组序列 广泛的研究人员与微生物多聚糖的相互作用。我们的假设是微生物多聚糖 填充有病原体和宿主微生物区系的阵列与相应的基因组序列相结合将 提供了一种独特且广泛有用的策略来定义宿主免疫因素对微生物的特异性 多聚糖。我们的假设是基于我们最近发现的先天免疫和获得性免疫 因子可以利用一个充满完整微生物的平台与各种不同的微生物多聚糖相互作用 以及它们相应的以微阵列形式印刷的微生物多聚糖。因为微生物群落是 非常多样化，我们的初步数据也表明，微生物可以从 复杂微生物混合物和微生物及其各自的葡聚糖可以类似地打印并 询问宿主免疫因子的相互作用。此外，从这些分离的基因中产生的基因组文库 微生物可以成功地融入到研究宿主微生物的传统方法中 互动，直接使该领域民主化。为了提供所需的微生物覆盖范围 有效地评估宿主微生物的相互作用，我们将在这些初步发现的基础上开发一套完整的 将通过以下具体目标向科学界广泛提供的工具： 目标1：开发集成的微生物多糖阵列和填充已知病原体的基因组数据库。 具体目标2：开发整合的微生物多糖阵列和填充了Normal的基因组数据库 微生物区系突出表现为与宿主免疫因子的相互作用。鉴于东道主的基本性质 与微生物多糖的相互作用以及阵列平台在使该领域“民主化”方面取得的有据可查的成功 在这项建议中开发的综合工具将为广泛的研究人员提供 有机会研究宿主-微生物的相互作用，对基础和疾病相关有重大影响 流程。

Summary: While successful immunity relies on the ability of host immune factors to rapidly recognize and respond to a variety of microbial determinants, microbial glycans often serve as the first and most important contact point with host immune factors. However, the binding specificity of most innate and adaptive immune factors toward microbial glycans remains largely unknown. Furthermore, despite the near universal use of genomic sequencing approaches to characterize microbiota, these methods largely fail to identify microbes based upon their unique glycan signatures, precluding the identification of distinct microbial strains with relevant glycan structures when using this approach. In order to effectively overcome current limitations in the study of host-microbial interactions, a platform must be developed that can be used by a wide variety of investigators to define how hosts interact with microbial glycans. Our long-term goal is to develop an integrated platform of microbial glycans and genomic sequences that can be used to define the network of innate and adaptive immune interactions with microbial glycans by a wide range of investigators. Our hypothesis is that microbial glycan arrays populated with pathogens and host microbiota coupled with corresponding genomic sequences will provide a unique and broadly useful strategy to define the specificity of host immune factors toward microbial glycans. Our hypothesis is formulated on the basis of our recent discoveries that innate and adaptive immune factors can interact with a variety of distinct microbial glycans using a platform populated with intact microbes and their corresponding microbial glycans printed in a microarray format. As microbial communities are extremely diverse, our preliminary data also demonstrate that microbes can be specifically isolated from a complex microbial mixture and that microbes and their respective glycans can be similarly printed and interrogated for host immune factor interactions. Furthermore, the genomic library generated from these isolated microbes can be successfully incorporated into traditional approaches designed to study host-microbial interactions, directly democratizing the field. In order to provide the breadth of microbial coverage needed to effectively assess host microbial interactions, we will build on these initial findings to develop an integrated set of tools that will be broadly available to the scientific community through the following specific aims: Specific Aim 1: Develop integrated microbial glycan arrays and genomic databases populated with known pathogens. Specific Aim 2: Develop integrated microbial glycan arrays and genomic databases populated with normal microbial flora highlighted by interactions with host immune factors. Given the fundamental nature of host interactions with microbial glycans and the documented success of array platforms in “democratizing” the field of glycosciences, the integrated tools developed in this proposal will provide a wide range of investigators the opportunity to study host-microbial interactions with significant implications on fundamental and disease-related processes.