Novel small molecule biosynthetic gene clusters in Streptococcus mutans and virulence of dental caries

变形链球菌新型小分子生物合成基因簇与龋齿毒力

• 批准号: 10378775
• 负责人: Stephanie Momeni
• 金额: $ 12.93万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 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 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10378775
• 关键词: 5 year old; 6 year old; Acids; Affect; Age; Animal Model; Appointment; Award; Bacteria; Bacterial Model; CISH gene; Career Choice; Child; Child Health; Child Welfare; Clinical; Collaborations; Communicable Diseases; Data; Dental Models; Dental Research; Dental caries; Development; Drosophila genus; Economic Burden; Epidemiology; Gene Cluster; Gene Expression; Genes; Genetic Transcription; Genome; Genotype; Health; Human; Hybrids; In Vitro; Ions; Journals; Knock-out; Knowledge; Lead; Life; Liquid Chromatography; Microbial Biofilms; Modeling; Molecular Biology; Mutagenesis; Mutation; Oral health; Pathogenesis; Phase; Phenotype; Population; Property; Publications; Publishing; RBM5 gene; Rattus; Recombinants; Research; Research Training; Risk; Role; Streptococcus mutans; Testing; Time; Virulence; Virulent; Work; antimicrobial peptide; base; biological adaptation to stress; butyrolactone; clinical phenotype; clinical translation; design; early childhood; environmental stressor; epidemiology study; fitness; fly; genome sequencing; health disparity; high risk; in vivo; insight; liquid chromatography mass spectrometry; metabolomics; microbial; multidisciplinary; mutant; new therapeutic target; novel; novel therapeutics; oral microbiome; prevent; programs; small molecule; study population; tandem mass spectrometry; targeted treatment; trait; transcriptome sequencing; transcriptomics; whole genome

Project Summary Dental caries is a global infectious disease impacting the lives of 80% of the human population. Severe early childhood caries (SECC) is of particular concern and characterized by extensive tooth decay that can dramatically affect the dental health of children under six years of age. This constitutes a significant economic burden, can have dramatic impact on the child’s well being and may contribute to overall health later in life. Streptococcus mutans is the bacterium most commonly associated with initiation of SECC and early childhood caries (ECC). Biosynthetic gene clusters (BGCs) in bacteria often encode genes for small molecules and other antimicrobial peptides. This research plan proposes to investigate a novel BGC (butyrolactone-ladderane hybrid, BL-BGC) of S. mutans from a high caries risk population to identify its impact on S. mutans virulence and survival traits. Preliminary data suggests the BL-BGC is significantly up regulated in dental caries and occurs in the most prevalent strain type of S. mutans within a high-caries risk study population. The hypothesis is that clinical S. mutans strains with the BL-BGC will have increased virulence properties and cariogenicity. Aim 1 will characterize phenotypes of clinical S. mutans strains containing the BL- BGC and their association with dental caries key virulence and survival traits using gene mutagenesis and in-vivo biofilm analysis. Aim 2 will determine the impact of BL-BGC on expression of virulence genes and metabolites in S. mutans biofilms using RNA-sequencing and metabolomics analysis. Aim 3 will determine the how BL-BGC impacts S. mutans colonization and virulence using animal models of dental caries. The proposed study will elucidate the role of BL-BGC in S. mutans virulence and fitness during the K99 phase. Metabolites produced by S. mutans with BL-BGC impacting key virulence traits will be purified and characterized in the R00 phase, providing new insights into S. mutans’ role in the pathogenesis of dental caries. Characterization of this and other BGCs may lead to new therapeutic targets designed to reduce, reverse, or even prevent SECC. The K99 phase of this award will provide the time and support for additional research training, building effective collaborations, enhancing professional development and submission of publications leading to a successful transition to an independent academic research appointment in the R00 phase. This study is unique in the use of BGCs from clinical S. mutans isolates from a high-risk caries population allowing for clinical translation of results and the use of metabolomics for identification of BL-BGC related compounds/metabolites. This study will enable an independent career path by establishing a multidisciplinary program utilizing molecular biology approaches to validate epidemiological associations, bridging these fields and advancing the knowledge of S. mutans virulence in caries.

项目摘要 牙科车是一种影响80％人口的生活的全球感染性疾病。 严重的幼儿汽车（SECC）特别关注，并以广泛的牙齿​​为特征 衰减会极大地影响六岁以下儿童的牙齿健康。这 构成巨大的经济燃烧，可能会对孩子的福祉产生巨大影响 可能在以后的生活中有助于整体健康。链球菌突变是细菌最常见的 与SECC和幼儿汽车（ECC）的启动有关。生物合成基因簇 （BGC）在细菌中通常编码小分子和其他抗菌胡椒的基因。这 研究计划的提案要研究一种新型的BGC（丁乙烯酮 - 甲烷杂种，BL-BGC） 来自高的葡萄链球菌具有风险人群，以识别其对链球菌病毒和生存的影响 特质。初步数据表明，BL-BGC在龋齿中受到了显着调节，并且发生 在最普遍的菌株类型中，在高中风险研究人群中。这 假设是，具有BL-BGC的临床链球菌菌株将具有增加的病毒特性 和致富性。 AIM 1将表征包含Bl-的临床链球菌菌株的表型 BGC及其与牙齿的关联使用基因诱变具有关键病毒和生存特征 和体内生物膜分析。 AIM 2将确定BL-BGC对病毒表达的影响 使用RNA序列和代谢组学分析，Mutans生物膜中的基因和代谢产物。目的 3将确定BL-BGC使用动物模型如何影响链球菌定植和病毒 龋齿。拟议的研究将阐明BL-BGC在链球菌病毒中的作用，并 在K99阶段的健身。由S. mutans产生的代谢产物具有BL-BGC撞击钥匙 病毒特征将在R00阶段进行纯化和表征，从而提供新的见解 S. Mutans在龋齿的发病机理中的作用。这个和其他BGC的表征可能 导致旨在减少，逆转甚至防止SECC的新治疗靶标。 K99阶段 该奖项将为其他研究培训提供时间和支持，建立有效 合作，增强专业发展和提交出版物，导致 成功过渡到R00阶段的独立学术研究任命。这项研究 在使用来自高危龋齿种群的临床链球菌分离株的BGC中是独一无二的 允许结果的临床翻译和代谢组学识别BL-BGC 相关化合物/代谢产物。这项研究将通过建立一个独立的职业道路 利用分子生物学方法验证流行病学的多学科计划 关联，桥接这些领域并促进携带中的Mutans病毒的知识。