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: 10215214
• 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 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10215214
• 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 龋病是一种全球性传染病，影响着80%的人类人口的生活。 严重的儿童早期龋病(SECC)特别令人关注，其特点是广泛的牙齿。 龋齿会极大地影响六岁以下儿童的牙齿健康。这 构成了重大的经济负担，可能对儿童的福祉产生重大影响， 可能有助于晚年的整体健康。变形链球菌是最常见的细菌 与SECC和早期儿童龋齿(ECC)的启动有关。生物合成基因簇 细菌中的(BGC)通常编码小分子和其他抗菌肽的基因。这 研究计划建议研究一种新型的丁内酯-梯烷杂化(BGC) 来自龋病高危人群的变形链球菌，以确定其对变形链球菌毒力和生存的影响 特征。初步数据显示，BL-BGC在龋齿中显著上调，并发生在 在高龋病风险研究人群中最流行的变形链球菌菌株类型。这个 假设具有BL-BGC的变形链球菌临床菌株将具有更强的毒力特性 和致龋性。目的1鉴定含有BL1基因的变形链球菌临床菌株的表型。 利用基因突变研究BGC及其与龋病关键毒力和生存性状的关系 和体内生物膜分析。目的2确定BL-BGC对毒力表达的影响 应用rna测序和代谢组学分析变形链球菌生物膜中的基因和代谢物。目标 3将通过动物模型确定BL-BGC如何影响变形链球菌的定植和毒力 有很多龋齿。这项拟议的研究将阐明BL-BGC在变形链球菌毒力和 在K99阶段的体能。变异链霉菌产生的代谢产物与BL-BGC的作用键 毒力特性将在R00阶段得到提纯和表征，为 变形链球菌在龋病发病机制中的作用该BGC和其他BGC的特征可以 导致旨在减少、逆转甚至预防SECC的新的治疗靶点。K99阶段 将为额外的研究培训提供时间和支持，建立有效的 合作，加强专业发展和提交出版物，从而 在R00阶段成功过渡到独立的学术研究任命。本研究 在使用来自高危龋齿人群的临床变异链球菌的bgc方面是独一无二的。 允许临床翻译结果和使用代谢组学来鉴定BL-BGC 相关化合物/代谢物。这项研究将通过建立一个 利用分子生物学方法验证流行病学的多学科计划 联系，沟通这些领域，并促进对变形链球菌在龋病中毒力的了解。