Oral microbial community structure and assembly: from molecule to microbiome

口腔微生物群落结构和组装：从分子到微生物组

• 批准号: 10447147
• 负责人: Alex M Valm
• 金额: $ 35.75万
• 依托单位: STATE UNIVERSITY OF NEW YORK AT ALBANY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-07 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10447147
• 关键词: Activities of Daily Living; Address; Anaerobic Bacteria; Apical; Bacterial Adhesins; Binding; Biochemical; Cell Communication; Cells; Coculture Techniques; Communities; Complement; Corynebacterium; Dental Plaque; Development; Disease; Ecosystem; Escherichia coli; Event; Filament; Fusobacterium; Fusobacterium nucleatum; Genes; Genetic; Gingiva; Gingivitis; Growth; Health Promotion; Health Transition; Heterogeneity; Image; In Vitro; Inflammatory; Knowledge; Leptotrichia; Location; Maps; Mediating; Membrane Proteins; Messenger RNA; Metagenomics; Microbe; Microbial Biofilms; Modeling; Molecular; Oral; Oral cavity; Organ; Organism; Oxygen; Pathogenicity; Pathway interactions; Pattern; Periodontal Diseases; Periodontitis; Phase; Phenotype; Play; Process; Property; Proteins; Publishing; Research; Resolution; Role; Seeds; Spatial Distribution; Structure; Structure-Activity Relationship; Surface; Taxonomy; Testing; Tissues; Transcript; Work; healthy volunteer; imaging study; imaging system; metagenomic sequencing; microbiome; mutant; neutralizing antibody; novel; oral biofilm; oral microbial community; prevent; receptor; synthetic biology; theories; tooth surface; transcriptome sequencing

ABSTRACT Just as the function of metazoan tissues and organs is mediated by the patterned assembly of phenotypically distinct cells during development, the functions of oral microbes in promoting health and disease arise from the emergent properties of their patterned structure: the biofilm. For example, the accumulation of supragingival dental plaque into stable, long-range biofilms at the gingival margin is implicated in reversible gingivitis, which may progress to chronic periodontitis. Recent studies have called into question the relative contribution of two conspicuous, filamentous organisms in structuring supragingival plaque over long distances: Fusobacterium nucleatum and Corynebacterium matruchotii. There exists a critical need to understand the role that filamentous organisms play in biofilm assembly because it is well understood that changes in taxonomic abundance, ecological succession and biofilm structure mediate the transition from health to inflammatory periodontal diseases in the mouth. To address this knowledge gap, we have developed an in vitro oral biofilm model, amenable to systems imaging in which to study the molecular basis of long-range community structure and biochemical interaction. We will elucidate the role of micron-scale spatial structure in oral biofilm assembly, growth and function, through combined metagenomic sequencing and systems imaging of biofilms inoculated with dental plaque from healthy donors. We will further map novel Corynebacterium genes involved in long range spatial structure of biofilms, through co-culture RNA-seq and mRNA FISH imaging. We will next decipher the specific F. nucleatum molecular components that underly long-range biofilm structure, through targeted inhibition of F. nucleatum outer membrane proteins with neutralizing antibodies and through genetic complementation of these proteins in E. coli cells co-cultured with dental plaque inocula. The knowledge gained from the research proposed here will comprise a holistic understanding of oral community assembly processes and physical structure across spatial scales from molecule to microbiome and may identify new paradigms for intervening in the progression of dental plaque-mediated diseases.

抽象的 正如后生动物组织和器官的功能是由表型的模式组装介导的一样 口腔微生物在发育过程中具有不同的细胞，其促进健康和疾病的功能源于 它们的图案结构的新兴特性：生物膜。例如，龈上积聚 牙菌斑在牙龈边缘形成稳定的长距离生物膜与可逆性牙龈炎有关， 可能会发展为慢性牙周炎。最近的研究对两种因素的相对贡献提出了质疑 长距离构建龈上菌斑的显着丝状生物：梭杆菌 具核杆菌和马特鲁霍蒂棒杆菌。迫切需要了解以下角色： 丝状生物在生物膜组装中发挥作用，因为众所周知，分类学的变化 丰度、生态演替和生物膜结构介导从健康到炎症的转变 口腔中的牙周疾病。为了解决这一知识差距，我们开发了一种体外口腔生物膜 模型，适合系统成像，用于研究远程群落结构的分子基础 和生化相互作用。我们将阐明微米级空间结构在口腔生物膜组装中的作用， 通过结合宏基因组测序和接种生物膜的系统成像来观察生长和功能 带有来自健康捐赠者的牙菌斑。我们将进一步绘制涉及长链的新棒状杆菌基因图谱 通过共培养 RNA-seq 和 mRNA FISH 成像，研究生物膜的空间结构。我们接下来 破译长程生物膜结构下的特定 F. nucleatum 分子成分，通过 使用中和抗体并通过基因靶向抑制具核梭菌外膜蛋白 与牙菌斑接种物共培养的大肠杆菌细胞中这些蛋白质的互补。知识点 从此处提出的研究中获得的成果将包括对口头社区集会的整体理解 从分子到微生物组的跨空间尺度的过程和物理结构，并可能识别新的 干预牙菌斑介导的疾病进展的范例。