A novel mechanism of virulence control in Porphyromonas gingivalis

牙龈卟啉单胞菌毒力控制的新机制

• 批准号: 10441545
• 负责人: Jorge Frias-Lopez
• 金额: $ 66.65万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10441545
• 关键词: Abscess; Address; Adult; Anabolism; Animal Model; BCAR1 gene; Bacteria; Bacteriophages; Cell Line; Cell Survival; Cells; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Cobalamin; Complex; DNA; DNA Repair; Data; Development; Disease; Disease Progression; Epithelial Cells; Francisella; Gene Expression; Genes; Genetic; Gingiva; Goals; Health; Health Transition; Human; IL8 gene; Immune; Immune response; Immunity; Infection; Inflammation; Inflammatory; Innate Immune Response; Interleukin-1 beta; Iron; Knowledge; Libraries; Measures; Metabolism; Modeling; Molecular; Moths; Mus; Operon; Oral; Outcome; Pathogenicity; Periodontal Diseases; Periodontitis; Play; Porphyromonas gingivalis; Production; Proteins; Proteolysis; Pseudomonas aeruginosa; Publishing; Reporting; Research; Role; Sampling; Signal Transduction; Site; System; TLR2 gene; TLR4 gene; TNF gene; Testing; Therapeutic; Tissues; Toll-like receptors; Tooth structure; Viral; Virulence; Virulence Factors; Waxes; Work; base; bone loss; experimental study; gene function; human microbiota; in vivo; infancy; keratinocyte; macrophage; metatranscriptome; microbial; microbial community; mutant; neutrophil; novel; oral infection; pathogen; periodontopathogen; polymicrobial disease; prevent; programs; transcriptome

Abstract Porphyromonas gingivalis is a major pathogen of severe adult periodontitis, a polymicrobial disease caused by the coordinated action of a complex microbial community that leads to inflammation of tissues supporting the teeth. A central hurdle limiting progress in periodontal disease research is the paucity of information detailing microbial signals that correlate with clinical progression at a site from health to disease. Filling this void, we recently reported metatranscriptome findings of the microbial community from human clinical samples during periodontal disease progression and discovered that CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)-associated proteins in the periodontopathogen P. gingivalis were highly up-regulated only at those sites that progressed. CRISPRs-Cas systems are used by bacteria to prevent foreign DNA incorporation, as occurs with a viral attack. The goal of this research program is to understand the role that CRISPR-Cas systems have on virulence determinants of important periodontopathogens during disease progression. A comprehensive analysis of the mutants will provide information required to increase our understanding of not only CRISPR gene function, but also the contribution of these novel genes to virulence. To this end we propose the following Specific Aims: Aim 1. Identify targeted endogenous genes comparing transcriptome profiles of the wild-type and the mutants growing intracellularly. Aim 2. Determine the impact of CRISPR-associated genes on the innate immune host responses to P. gingivalis. Aim 3. Aim 3. Determine the role of CRISPR-Cas genes in the pathogenicity of P. gingivalis. We expect that this knowledge will facilitate the development of targeted approaches to prevent and treat periodontitis by inhibiting specific Cas proteins essential for virulence. Such results will fundamentally advance our understanding that such systems have in the metabolism of periodontal pathogens besides their traditional role assigned as a mechanism of protection against foreign DNA. We believe that the team we have assembled for this project has all the qualifications to accomplish successfully the goals proposed in the present application.

抽象的 牙龈卟啉单胞菌是严重成人牙周炎的主要病原体，这是一种由多种微生物引起的疾病 复杂微生物群落的协调作用，导致支持组织的炎症 牙齿。限制牙周病研究进展的一个主要障碍是缺乏详细信息 与从健康到疾病的临床进展相关的微生物信号。填补这个空白，我们 最近报道了人类临床样本中微生物群落的宏转录组发现 牙周病的进展，并发现 CRISPR（簇状规则间隔短 牙周病原菌 P. gingivalis 中的回文重复 (Palindromic Repeats) 相关蛋白仅高度上调 在那些取得进展的网站上。细菌利用 CRISPRs-Cas 系统来阻止外来 DNA 合并，就像病毒攻击时发生的那样。该研究计划的目标是了解 CRISPR-Cas系统对疾病期间重要牙周病原体的毒力决定因素 进展。对突变体的全面分析将提供增加我们所需的信息 不仅了解 CRISPR 基因功能，还了解这些新基因对毒力的贡献。到 为此，我们提出以下具体目标： 目标 1. 通过比较野生型和突变体的转录组图谱来识别目标内源基因 在细胞内生长。 目标 2. 确定 CRISPR 相关基因对宿主对疟原虫的先天免疫反应的影响。 牙龈。 目标 3. 目标 3. 确定 CRISPR-Cas 基因在牙龈卟啉单胞菌致病性中的作用。 我们期望这些知识将有助于制定有针对性的方法来预防和治疗 通过抑制对毒力至关重要的特定 Cas 蛋白来治疗牙周炎。这些成果将从根本上推进我们的 了解此类系统除了其传统作用外，还在牙周病原体的代谢中发挥作用 被指定为一种针对外来 DNA 的保护机制。我们相信我们为此组建的团队 该项目具有成功实现本申请中提出的目标的所有资格。