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.

项目总结/摘要 拟议工作的广泛目标是解决对细菌病毒作用缺乏了解的问题 （3）塑造口腔微生物群落的生态。噬菌体是重要的捕食者， 所有环境中的细菌共生体，并且它们在口腔中大量存在（唾液中高达108 ml-1 和107 mg-1斑块）。通过对细菌宿主的急性和慢性感染， 潜在的塑造细菌种群结构，定殖动力学和应变水平的变化， 毒性然而，尽管口腔微生物组中的细菌已经研究了几十年，但对它们的作用知之甚少。 如何塑造这些社区的结构和功能，从而在人类社会中发挥作用， 健康和疾病。在本申请中，一种细菌病原体已经并将继续- 卟啉单胞菌作为牙周病的关键菌种和驱动菌， 牙龈菌（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，尚未分离到能感染或杀死Pg的病原体，也没有系统地研究Pg基因组。 调查慢性感染形式的噬菌体（“原噬菌体”）的证据。的中心假设 所提出的工作是Pg分离物携带作为前噬菌体整合到其基因组中的不同的噬菌体， 这些原噬菌体有可能改变口腔微生物组中Pg的生态。两 互补的目标，提供了广度和深度的洞察力的作用，在生态学的 PG，用于解决这一假设。在目标1中，目标是识别和功能表征 这是通过从志愿者中获得新的Pg分离株来实现的 与牙周病，以扩大可用的Pg基因组的数量，并生物信息学识别 并描述这些基因组中的原噬菌体，包括它们促进 细菌毒力和种内竞争。在目标2中，目标是确定 感染它们的Pg宿主。这是通过建立Pg函数的模型系统， 以种植为基础的方法，以确定其宿主范围的决定因素。这些目标的实现将为 第一个观点的系统发育和功能多样性的Pg的，洞察他们的主机范围， 这是第一次鉴定出它们用来感染Pg宿主的受体。这将为研究提供 社区的知识基础和工具，必要的测试假设（在体外，在体内， 纵向和横截面研究），解决了如何相互作用与非线形Pg基因组， 生理学、种群动态、种间和王国间的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-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坚固、安全， 以及用于口腔和全身性疾病包括牙周病的有效的基于噬菌体的治疗剂。