Targeted Cultivation of New Periodontal Pathogens

新牙周病原菌的定向培养

• 批准号: 9561809
• 负责人: Flavia Rocha Teles
• 金额: $ 27.61万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-08 至 2018-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9561809
• 关键词: Adult; Affect; Animal Model; Bacteria; Biological; Cardiovascular system; Clinical; Communicable Diseases; Communities; Computer Simulation; Data; Development; Devices; Diagnosis; Disease; Elements; Environment; Etiology; Forsythia; Fostering; Generations; Genomics; Goals; Growth; Habitats; Health; Immune response; In Situ; In Vitro; Infection; Knowledge; Link; Microbial Biofilms; Minority; Mission; Modeling; Names; National Institute of Dental and Craniofacial Research; Oral; Organism; Outcome; Pathogenesis; Pathogenicity; Pathway Analysis; Periodontal Diseases; Periodontitis; Play; Porphyromonas gingivalis; Prevention; Procedures; Public Health; Research; Ribosomal RNA; Role; Sampling; Scientist; Testing; Tooth Loss; Treatment Cost; Virulence; Work; base; bone loss; cost effective; improved; innovation; interest; member; microbial community; microbiota; next generation sequencing; novel; oral infection; oral microbiome; pathogen; public health relevance; respiratory; subgingival biofilm; success; treatment response

DESCRIPTION (provided by applicant): Periodontitis affects 64 million US adults, leading to bone and tooth loss and contributing to systemic conditions. Subgingival bacteria play a major role in periodontitis. Because 50% of the oral microbiome is unrecognized or uncultured, important species may have been overlooked, representing a gap in knowledge. The progress in cultivating relevant oral uncultured taxa has been slow. The few studies available are either low-throughput or open-ended, which are less suitable to cultivate putative pathogens. Because our goal is to improve periodontal health, we aim at cultivating taxa that are relevant in disease, and not just any uncultured taxa. Results from my R03 showed that many unrecognized/uncultured subgingival taxa were uncommon, thus unlikely to be relevant. In contrast, I identified 30 abundant uncultured taxa that merit further pursuit and will be the focus of this proposal. Our long term goal is to identify new periodontal pathogens and their pathogenic mechanisms. The overall objective of this proposal is to recover and domesticate uncultured taxa associated with periodontal disease. Our central hypothesis is that their growth results from interactions with community members and the habitat, which have not yet been reproduced in vitro. Our rationale is that targeted systematic cultivation of relevant taxa in ex vivo biofilms is a cost-effective approach to isolate new periodontal pathogens. We will use technological and conceptual innovative approaches to achieve our goals. We will test this hypothesis and accomplish our objective by pursuing the following Specific Aims: 1) Support the growth of prominent uncultured taxa in ex vivo periodontal biofilms: we will use the Calgary Biofilm Device to multiplex the cultivation conditions (using multiple growth conditions in paralle) of subgingival samples pre-screened for the target taxa using 16S rRNA lllumina sequencing; 2) Identify "helper" species that can sustain the growth of target uncultured periodontal taxa: we will use CLASI-FISH (a multiplex FISH) on the biofilms and correlation network analysis of 16S rRNA data to identify in situ and in silico the best "helper" species based on their co-occurrence and physical proximity with target uncultured taxa; and 3) Domesticate "uncultured" periodontal taxa in a small consortium of helper species: we will use the best growth conditions and helpers species identified above for the high-throughput parallel co-cultivation of uncultured taxa. This proposal is significant because it will allow the cultivation of novel putative periodontal pathogens, a key step to determine their pathogenicity. Our contribution to the field will be the identification of ecological relationships that allow their growth, the generation of whole genomic sequences as well as the development of a systematic approach that can be useful to study other biofilm diseases. The outcomes of our study will support NIDCR's research initiatives for FY2014 (the role of uncultivable bacteria in the oral microbiome) and their mission to foster the knowledge of the etiology and pathogenesis of oral infectious diseases.

描述（由申请人提供）：牙周炎影响6400万美国成年人，导致骨和牙齿脱落并导致全身疾病。牙龈下细菌在牙周炎中起主要作用。由于50%的口腔微生物组未被识别或未被培养，重要的物种可能被忽视，这代表了知识上的差距。相关口腔未培养类群的培育进展缓慢。现有的少数研究要么是低通量的，要么是开放式的，这些研究不太适合培养假定的病原体。因为我们的目标是改善牙周健康，我们的目标是培养与疾病相关的分类群，