Impact of ß-glucan metabolism on the development of pathogenic biofilms

α-葡聚糖代谢对致病性生物膜形成的影响

• 批准号: 10057468
• 负责人: Kiyonobu Homma
• 金额: $ 15.95万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2022-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10057468
• 关键词: Advanced Glycosylation End Products; Affect; Alveolar Bone Loss; Applications Grants; Bacteria; Biomass; Candida; Carbon; Cells; Chronic; Communication; Communities; Complex; Data; Dental Plaque; Development; Diet; Disease; Enzymes; Forsythia; Foundations; Fusobacterium nucleatum; Future; Genetic Transcription; Glucans; Glucose; Goals; Gram-Negative Bacteria; Growth; Human; Hydrolase; Hydrolysis; In Vitro; Inflammation; Inflammatory; Investigation; Mediating; Metabolic; Metabolism; Microbial Biofilms; Modeling; Molecular; Mus; Nutrient; Odontogenesis; Operon; Oral; Pathogenesis; Pathogenicity; Periodontal Infection; Periodontitis; Physiology; Play; Poison; Production; Publications; Pyruvaldehyde; Regulation; Role; Sigma Factor; Source; Specificity; Stimulus; Structure; System; Tissues; Tooth Loss; Tooth structure; Virulence; Yeasts; alveolar bone; beta-Glucans; dental biofilm; dysbiosis; in vitro Model; inhibitor/antagonist; member; microbial; microbial community; mouse model; oral bacteria; oral microbial community; pathogen; polymicrobial biofilm; response; subgingival biofilm; therapy development; transcriptomics

Project Summary/Abstract Periodontitis is a bacterially induced chronic inflammation of the tooth supporting tissues that may result in alveolar bone destruction and tooth loss. Tannerella forsythia (Tf), a member of the ‘red-complex’ bacteria group, is one of the major pathogens implicated in periodontitis. T. forsythia has been shown to co-aggregate with the ‘bridge-bacterium’ Fusobacterium nucleatum (Fn) to form synergistic co-biofilms in vitro and induce alveolar bone loss in mice when co- infected with Fn. The overall goal of this proposal is to determine the molecular mechanisms of T. forsythia -F. nucleatum intergeneric interactions to better understand the pathogenesis of periodontitis. Our preliminary data show that a Tf secreted ß-glucanase enzyme (GlcA) whose expression is induced in response to Fn sensing plays a significant role in the development of mixed biofilms. We showed that this enzyme hydrolyzes ß-glucans into glucose, which serves as a nutrient for Fn to promote its biomass in Tf -Fn co-biofilms. Our data showed that the increased glucose availability did not affect Tf biomass but it rather enhanced the production of methylglyoxal (MGO), a highly reactive dicabonyl compound toxic to bacterial and host cells. The goal of this proposal is to understand the mechanistic basis of Tf-Fn interactions and how metabolic interactions between these two species contribute to the development of the dental plaque, dysbiosis and inflammation. Our working hypothesis is that b-glucanase produced by Tf in response to Fn, and possible other stimuli, releases glucose from dietary b- glucans as a nutrient for the microbial community at large and as a metabolic precursor for MGO secretion to favor microbial dysbiosis. To interrogate this hypothesis, we propose two specific aims: Aim 1: To define the molecular mechanism of regulation of Tf GlcA operon in response to Fn sensing. We will analyze the mechanism and function of ECF sigma-anti sigma system that is predicted to drive Tf glcA ß-glucanase operon to respond stimulate with Fn and other stimuli, and; Aim 2 To determine the mechanisms by which Fn resists Tf-produced methylglyoxal and how hydrolyzed glucans impact microbial community structure. Here, we will determine the molecular mechanisms by which Fn detoxifies MGO produced by Tf in biofilms and promote dental plaque development. Successful completion of this study will form the foundation for future investigations exploring the unique ability of Tf glucanase-MGO axis in promoting microbial dysbiosis and subsequently develop glucanase targeting inhibitors to block the synergistic Tf-Fn associations and dental plaque development for the treatment of periodontitis.

项目概要/摘要 牙周炎是细菌引起的牙齿支持组织的慢性炎症 这可能会导致牙槽骨破坏和牙齿脱落。连翘坦尼氏菌 (Tf)，成员 “红色复合体”细菌群的一员，是与牙周炎有关的主要病原体之一。 T。 连翘已被证明与“桥梁细菌”具核梭杆菌共同聚集 (Fn) 在体外形成协同生物膜并诱导小鼠牙槽骨丢失 感染Fn。该提案的总体目标是确定其分子机制 连翘-F。 nucleatum 属间相互作用，以更好地了解其发病机制 牙周炎。我们的初步数据表明，Tf 分泌 β-葡聚糖酶 (GlcA)，其 响应 Fn 感应而诱导表达，在混合细胞的发育中发挥重要作用 生物膜。我们发现这种酶可将 ß-葡聚糖水解成葡萄糖，而葡萄糖可作为 Fn 的营养物质 促进其在 Tf-Fn 共生物膜中的生物量。我们的数据表明，葡萄糖利用率的增加并没有 影响 Tf 生物量，但它反而增强了甲基乙二醛 (MGO) 的产生，甲基乙二醛是一种高反应性二碳基 对细菌和宿主细胞有毒的化合物。 该提案的目标是了解 Tf-Fn 相互作用的机制基础 以及这两个物种之间的代谢相互作用如何促进 牙菌斑的形成、菌群失调和炎症。我们的工作假设是 Tf 响应 Fn 和可能的其他刺激而产生的 b-葡聚糖酶，从膳食 b- 中释放葡萄糖 葡聚糖作为整个微生物群落的营养物质和 MGO 分泌的代谢前体 有利于微生物失调。为了质疑这一假设，我们提出了两个具体目标： 目标 1： 定义响应 Fn 传感的 Tf GlcA 操纵子调节的分子机制。 我们将分析所预测的ECF sigma-anti sigma系统的机制和功能 驱动 Tf glcA ß-葡聚糖酶操纵子对 Fn 和其他刺激作出反应；目标2 确定 Fn 抵抗 Tf 产生的甲基乙二醛的机制以及如何 水解葡聚糖影响微生物群落结构。在这里，我们将确定分子 Fn 解毒生物膜中 Tf 产生的 MGO 并促进牙菌斑的机制 发展。 本研究的成功完成将为今后的研究奠定基础 Tf 葡聚糖酶-MGO 轴在促进微生物失调并随后发展方面的独特能力 葡聚糖酶靶向抑制剂可阻断 Tf-Fn 协同作用和牙菌斑 开发用于治疗牙周炎。