Molecular analysis of F. nucleatum interspecies interactions in biofilms

生物膜中具核梭菌种间相互作用的分子分析

基本信息

批准号：
8495763
负责人：
Renate Lux
金额：
$ 36.96万
依托单位：
UNIVERSITY OF CALIFORNIA LOS ANGELES
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-07-11 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8495763
关键词：
Acute Address Adherence Adhesions Animal Model Bacteria Bacterial Adhesins Behavior Binding Cells Chronic Communicable Diseases Communities Community Integration Complex Development Event Fusobacterium nucleatum Future Gene Expression Profile Gene Expression Regulation Gene Fusion Gene Silencing Genes Genetic Gingival Pocket Goals Human In Vitro Infection Investigation Knowledge Laboratories Libraries Luciferases Mediating Membrane Proteins Microbial Biofilms Modeling Molecular Molecular Analysis Molecular Target Open Reading Frames Operon Oral Oral cavity Organism Outcome Pathogenesis Pattern Periodontal Diseases Periodontitis Physiological Play Process Property Proteins Publishing Reporter Reporter Genes Research Role Signal Transduction Solid Streptococcus Structure Surface System Testing Therapeutic Therapeutic Intervention Time Tissue-Specific Gene Expression Tissues Tooth Loss Virulence Work base fascinate genetic manipulation improved insight luminescence mutant novel oral behavior oral biofilm oral tissue pathogen pathogenic bacteria research study tool tooth surface

项目摘要

DESCRIPTION (provided by applicant): The oral cavity harbors a complex microbial biofilm community capable of mediating acute and chronic polymicrobial infections including periodontal diseases which are still the most common reason for tooth loss worldwide. Specific interspecies interactions are key factors in constructing these communities and Fusobacterium nucleatum (Fn) has been established as the central "bridging" organism that enables arrangement of numerous oral bacterial species (both "early" and "late" colonizers) into well organized communities for biofilm formation and pathogenesis. The ability of Fn to adhere to a large variety of oral species has been clearly demonstrated through elaborate in vitro co-aggregation studies with planktonic cells, yet only limited studies have explored how this ability manifests in multispecies microbial biofilms. More importantly, lack of effective genetic manipulation tools and clear molecular targets has hampered investigation of the molecular mechanisms behind these Fn-mediated fascinating interspecies interactions. We recently achieved the first genetic characterization of a fusobacterial adhesin (RadD) that mediates interspecies adhesion with early oral colonizers. In our preliminary studies, we demonstarted that RadD- dependent binding to its partner species Streptococcus sanguinis (Ss) stimulates distinct phenotypic changes and differential gene expression in Fn. Based on these findings, we developed our working hypothesis that "Fn interacts via RadD and associated Rad-proteins with early colonizing partner species, which triggers significant morphological and physiological changes in Fn important for biofilm formation". To test this hypothesis, we propose a detailed examination of the role of RadD in Fn-mediated interspecies interactions using Streptococcus sanguinis (Ss) as model early colonizing partner species. Approaches will include characterization of biofilm behavior, oral community integration, gene expression patterns and identification of cellular components in Ss involved in the OMP-mediated interaction with Fn. This comprehensive approach will provide detailed knowledge at the molecular level for the events involved in biofilm formation including physical cell-cell contact and signaling. This research will provide novel targets for improved therapeutic intervention of a widespread human infectious disease and serve as a basis for future studies of Fn with late colonizing partner species or animal models.

描述（由申请人提供）：口腔含有一个复杂的微生物生物膜群落，能够介导急性和慢性多细胞感染感染，包括牙周疾病，这仍然是全球最常见的牙齿脱落原因。特定种间相互作用是构建这些群落的关键因素，核细菌核（FN）已被确定为中心的“桥接”生物体，可以使许多口腔细菌物种（包括“早期”和“晚期”殖民者）在生物膜形成和发病机理中排列众多口腔细菌（包括“早期”和“晚期”殖民者）。通过与浮游细胞的详尽的体外共聚集研究，清楚地证明了FN粘附在各种口腔物种上的能力，但只有有限的研究探索了这种能力如何在多种质子微生物生物膜中表现出来。更重要的是，缺乏有效的遗传操纵工具和清晰的分子靶标，这阻碍了对这些FN介导的引人入胜的种间相互作用背后的分子机制的研究。我们最近实现了融合蛋白（RADD）的第一个遗传表征，该粘蛋白（RADD）介导了与早期口服菌落的种间粘附。在我们的初步研究中，我们揭示了与其伴侣物种链球菌（SS）的radd依赖性结合刺激了FN中不同的表型变化和差异基因表达。基于这些发现，我们开发了我们的工作假设：“ FN通过RADD和相关的Rad蛋白与早期定植伴侣物种相互作用，这触发了FN对生物膜形成重要的形态和生理变化的显着变化”。为了检验这一假设，我们提出了对RADD在FN介导的种间相互作用中使用链球菌（SS）作为模型早期定居伴侣物种的作用的详细研究。方法将包括表征生物膜行为，口腔群落整合，基因表达模式以及涉及与FN相互作用的SS中细胞成分的鉴定。这种全面的方法将在分子水平上为生物膜形成的事件提供详细的知识，包括物理细胞 - 细胞接触和信号传导。这项研究将为改善广泛的人类传染病的治疗干预提供新的靶标，并为未来与晚定殖民伴侣物种或动物模型的FN进行研究。