Uncovering the roles of phages in the ecology of Porphyromonas gingivalis in periodontal disease

揭示噬菌体在牙周病牙龈卟啉单胞菌生态学中的作用

• 批准号: 10528068
• 负责人: Kathryn M Kauffman
• 金额: $ 16万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT BUFFALO
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-07-01 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10528068
• 关键词: Address; Animal Model; Bacteria; Bacteriophages; Bioinformatics; Biological; Biological Models; Chronic; Clinical; Clinical Research; Clustered Regularly Interspaced Short Palindromic Repeats; Collection; Communities; Cross-Sectional Studies; Data Set; Development; Disease; Disease Progression; Ecology; Environment; Exposure to; Foundations; Future; Genome; Genomics; Goals; Health; Human; In Vitro; Individual; Infection; Information Systems; Knowledge; Link; Longitudinal Studies; Mediating; Methods; Microbe; Mouth Diseases; Oral; Oral cavity; Periodontal Diseases; Phylogenetic Analysis; Physiology; Play; Population; Population Dynamics; Porphyromonas gingivalis; Predatory Behavior; Prophages; Research; Resistance; Resources; Role; Saliva; Sampling; Sampling Studies; Shapes; Site; Source; Structure; Study models; Surface; System; Systemic disease; Technology; Testing; Therapeutic Uses; Variant; Virulence; Virulence Factors; Virus; Work; acute infection; base; capsule; chronic infection; cohort; extracellular; in vivo; insight; knowledgebase; mutant; nanopore; novel therapeutics; oral bacteria; oral microbial community; oral microbiome; particle; pathogen; pathogenic bacteria; pressure; rational design; receptor; screening; symbiont; tool; volunteer

PROJECT SUMMARY/ABSTRACT The broad goal of the proposed work is to address the lack of knowledge about the roles of bacterial viruses (phages) in shaping the ecology of oral microbial communities. Phages are important predators and symbionts of bacteria in all environments, and they are abundant in the oral cavity (up to 108 ml-1 in saliva and 107 mg-1 plaque). Through acute and chronic infections of their bacterial hosts, phages have the potential to shape bacterial population structure, colonization dynamics, and strain level variation in virulence. Yet, though bacteria in the oral microbiome have been studied for decades, little is known about how phages shape the structure and function of these communities and thereby may play a role in human health and disease. In this application, a bacterial pathogen that has been – and continues to be - intensively studied for its role as keystone species and driver in periodontal disease, Porphyromonas gingivalis (Pg), is developed as a model for studying bacteria-phage interactions in the oral microbiome. To date, no phages able to infect or kill Pg have been isolated, nor have Pg genomes been systematically investigated for evidence of chronically infecting forms of phages (“prophages”). The central hypothesis of the proposed work is that Pg isolates harbor diverse phages integrated into their genomes as prophages, and that these prophages have the potential to alter the ecology of Pg in the oral microbiome. Two complementary aims, providing both breadth and depth of insight into the roles of phages in the ecology of Pg, are used to address this hypothesis. In Aim 1, the goal is to identify and functionally characterize prophages encoded in genomes of Pg. This is achieved by obtaining new isolates of Pg from volunteers with periodontal disease to expand the number of available Pg genomes, and bioinformatically identifying and characterizing prophages in these genomes, including with respect to their capacity to contribute to bacterial virulence and intraspecies competition. In Aim 2, the goal is to determine the receptors used by prophages to infect their Pg hosts. This is achieved by establishing model systems of Pg phages and using cultivation based methods to identify their host range determinants. Completion of these aims will provide the first view of the phylogenetic and functional diversity of Pg phages, insights into their host ranges, and the first identification of receptors that they use to infect their Pg hosts. This will provide the research community with the knowledgebase and tools necessary to test hypotheses (in vitro, in vivo, and in longitudinal and cross-sectional studies) that address how interactions with phages shape Pg genomes, physiology, population dynamics, inter-species and inter-kingdom interactions, and role in periodontal disease and its progression. Uncovering fundamental principles of how phage-bacteria interactions are structured in the oral microbiome will also lay an essential foundation for the rational design of robust, safe, and efficient phage-based therapeutics for use in oral and systemic disease including periodontal disease.

项目概要/摘要 拟议工作的总体目标是解决对细菌病毒的作用缺乏了解的问题 （噬菌体）塑造口腔微生物群落的生态。噬菌体是重要的捕食者 细菌在所有环境中都有共生体，并且在口腔中含量丰富（唾液中高达108ml-1） 和 107 mg-1 斑块）。通过细菌宿主的急性和慢性感染，噬菌体具有 塑造细菌种群结构、定植动态和菌株水平变化的潜力 毒力。然而，尽管口腔微生物组中的细菌已经被研究了几十年，但人们对它们知之甚少。 噬菌体如何塑造这些群落的结构和功能，从而在人类中发挥作用 健康和疾病。在此应用中，一种细菌病原体已经并且将继续是- 对其作为牙周病关键物种和驱动因素的作用进行了深入研究，卟啉单胞菌 gingivalis (Pg) 被开发为研究口腔微生物组中细菌-噬菌体相互作用的模型。到 迄今为止，还没有分离出能够感染或杀死 Pg 的噬菌体，也没有系统地研究过 Pg 基因组 调查了慢性感染形式的噬菌体（“原噬菌体”）的证据。中心假设为 拟议的工作是 Pg 分离株将多种噬菌体作为原噬菌体整合到其基因组中， 这些原噬菌体有可能改变口腔微生物组中 Pg 的生态。二 互补的目标，提供了对噬菌体在生态学中的作用的广度和深度的洞察 Pg，用于解决这个假设。目标 1 的目标是识别并功能表征 Pg 基因组中编码的前噬菌体。这是通过从志愿者那里获得新的 Pg 分离株来实现的 与牙周病一起扩大可用 Pg 基因组的数量，并通过生物信息学方法识别 并表征这些基因组中的前噬菌体，包括它们贡献的能力 细菌毒力和种内竞争。在目标 2 中，目标是确定所使用的受体 噬菌体感染其 Pg 宿主。这是通过建立 Pg 噬菌体模型系统并使用 基于栽培的方法来确定其宿主范围决定因素。完成这些目标将提供 首次了解 Pg 噬菌体的系统发育和功能多样性，了解其宿主范围，以及 首次鉴定出它们用来感染 Pg 宿主的受体。这将为研究提供 拥有测试假设（体外、体内和体内）所需的知识库和工具的社区 纵向和横断面研究）解决与噬菌体的相互作用如何塑造 Pg 基因组， 生理学、人口动态、物种间和王国间相互作用以及在牙周病中的作用 疾病及其进展。揭示噬菌体与细菌相互作用的基本原理 口腔微生物组的结构也将为合理设计稳健、安全、 以及有效的基于噬菌体的疗法，用于治疗口腔和全身疾病，包括牙周病。