Exploring the Ecological Role of Rothia mucilaginosa and It's Iron Binding Siderophore Enterobactin
探索 Rothia mucilaginosa 及其铁结合铁载体肠杆菌素的生态作用
基本信息
- 批准号:10366025
- 负责人:
- 金额:$ 29.25万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2021
- 资助国家:美国
- 起止时间:2021-03-05 至 2024-02-29
- 项目状态:已结题
- 来源:
- 关键词:3-DimensionalActinobacteria classAddressAffinityAnti-Inflammatory AgentsBackBacteriaBindingBiochemical ProcessBiological AssayBiological ModelsBody FluidsCarrier ProteinsCatecholsCell LineCell SeparationCellsChelating AgentsCoculture TechniquesCommunitiesComplexConfocal MicroscopyDental PlaqueDoseEcologyEnterobactinEnvironmentEnzymesEpithelial CellsEscherichia coliExposure toFerritinFlow CytometryFluorescence-Activated Cell SortingFoundationsFree RadicalsFutureGene ClusterGenesGenetic TranscriptionGenomeGoalsGrowthGut MucosaHumanHuman MicrobiomeHuman bodyImmune systemIn VitroIncubatedIndividualInterdisciplinary StudyIronIron Chelating AgentsKnowledgeLabelLifeLightMeasurementMeasuresMetabolicMicrobial BiofilmsMicroscopyMolecularMonitorMucous MembraneOralOral cavityOral healthOral mucous membrane structureOrganismOutcomeOxidative StressPathogenicityPhenotypePlayPopulationProductionProteinsPublishingReactive Oxygen SpeciesReporterReporter GenesResearchRhodamineRoleSalivaSentinelSiderophoresSignal TransductionStreptococcus mutansStreptococcus salivariusSystemTaxonomyTestingTissuesTransferrinWorkbacterial communitycommensal bacteriacomparativecytokineexperimental studyextracellulargut microbiotahost microbiotahost-microbe interactionshuman modelin vivointerestiron metabolismmembermetagenomic sequencingmetatranscriptomicsmicrobialmicrobial hostmicrobiome researchmultiple omicsnasal microbiotaoral bacteriaoral cavity epitheliumoral commensaloral microbial communityreceptorresponserhosmall moleculethree-dimensional modelingtranscriptomics
项目摘要
Project Summary
The oral cavity is a highly diverse microbial environment, consisting of >2000 bacterial, archaeal, and fungal
species most of which have not been functionally characterized. Numerous studies have identified that oral
Rothia mucilaginosa, a Gram-positive oral commensal Actinobacteria, is highly abundant in saliva and dental
plaque in global human populations, however, its ecological role is unknown. We identified that R. mucilaginosa
produced the catechol siderophore enterobactin, the strongest iron-chelating molecule known. We also identified
the enterobactin biosynthetic gene cluster (ent-BGC) in global Rothia genomes, which suggests that enterobactin
is crucial in Rothia ecology. The purified enterobactin compound impacted growth differentially when amended
to cultures of other oral bacterial species. It boosted the growth of commensal Streptococcus salivarius while it
reduced the growth of some strains of pathogenic S. mutans. The overarching goal of this study is to determine
the role of R. mucilaginosa produced enterobactin in interactions with both the oral microbiota and human oral
epithelial cells representing the oral mucosa. We propose an interdisciplinary research approach with two
specific aims: Aim 1: Characterization of molecular and ecological responses of oral bacteria to R.
mucilaginosa produced enterobactin. The activities of enterobactin will be characterized both in highly diverse
oral in vitro grown biofilms, and in bacterial monocultures. The capacity to import enterobactin by different
bacterial community members will be revealed by amending growth cultures with fluorescently rhodamine-
labeled enterobactin, fluorescence-activated cell sorting, and confocal microscopy. Multi-OMICS sequencing will
be conducted to characterize functional changes both in biofilm communities as well as in single and dual-
species cultures, specifically targeting genes encoding transport proteins and release mechanisms of
enterobactin. Aim 2: Determine the impact of enterobactin on oral mucosal sentinel cells. Co-cultivation
systems including both 2D and 3D models of human oral epithelial cells will be applied to study interactions with
the oral microbiota and enterobactin. Outcomes of the interactions will be characterized by using a multipronged
approach including a comparative gene transcription approach and a gene reporter system that reveals ROS
activation in host cells. Production of extracellular pro- and anti-inflammatory cytokines in cell medium relation
to enterobactin and oral bacteria will also be addressed to elucidate interactions of importance.
The proposed study provides a unique opportunity to expand our knowledge on enterobactin functional role in
the oral microbiota, and in interactions with human oral epithelial cells, which is severely lacking. A deeper
knowledge of the role of iron scavenging siderophores in the oral cavity will likely bring about a shift in the
research field of oral microbial ecology and shine a new light on the importance of iron metabolism in oral health.
项目摘要
口腔是一个高度多样化的微生物环境,由细菌、古生菌和真菌组成。
其中大多数还没有功能特征的物种。许多研究已经证实,口头的
粘性罗氏菌是一种革兰氏阳性口腔共生放线杆菌,在唾液和牙齿中含量非常丰富
然而,斑块在全球人群中的生态作用尚不清楚。我们鉴定出粘质乳杆菌
产生了儿茶酚铁载体Enterobactin,这是已知的最强的铁络合分子。我们还确认了
在全球Rothia基因组中发现了Enterobactin生物合成基因簇(ent-BGC),这表明Enterobactin
在Rothia生态学中是至关重要的。纯化的肠杆菌素化合物对生长发育的影响不同
其他口腔细菌种类的培养。促进共生性唾液链球菌的生长,同时
抑制某些致病性变形链球菌的生长。这项研究的首要目标是确定
粘质乳杆菌产生的肠杆菌素在与口腔微生物区系和人类口腔相互作用中的作用
代表口腔粘膜的上皮细胞。我们提出了一种跨学科的研究方法,包括两个
具体目标:目标1:口腔细菌对R。
粘菌产生肠杆菌素。Enterobactin的活性将具有高度多样性的特点
口腔体外培养的生物膜,以及细菌单一培养。不同品种进口肠毒素的能力
细菌群落成员将通过用荧光罗丹明修饰生长培养物来揭示-
标记肠动蛋白、荧光激活细胞分选和共聚焦显微镜。多OMICS测序将
对生物膜群落以及单细胞和双细胞生物膜群落的功能变化进行表征。
物种培养,特别针对编码运输蛋白的基因和释放机制
肠抗菌素。目的2:探讨肠杆菌素对口腔黏膜前哨细胞的影响。共育
包括人类口腔上皮细胞2D和3D模型的系统将被应用于研究与
口腔微生物区系和肠道菌素。互动的结果将通过使用多管齐下的
包括比较基因转录方法和揭示ROS的基因报告系统的方法
宿主细胞中的激活。细胞培养上清液中细胞外促炎症和抗炎细胞因子的产生
还将阐述肠杆菌素与口腔细菌相互作用的重要性。
这项拟议的研究提供了一个独特的机会来扩大我们对Enterobactin在
口腔微生物区系,以及与人类口腔上皮细胞的相互作用,这是严重缺乏的。更深一层的
对口腔中铁清除铁载体的作用的了解可能会带来口腔中
了解口腔微生物生态学的研究领域,使人们重新认识到铁代谢在口腔健康中的重要性。
项目成果
期刊论文数量(4)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Piloting a city health adaptation typology with data from climate-engaged cities: Toward identification of an urban health adaptation gap.
利用气候参与城市的数据试点城市健康适应类型:确定城市健康适应差距。
- DOI:10.1016/j.envres.2020.110435
- 发表时间:2021
- 期刊:
- 影响因子:8.3
- 作者:Sheehan,MaryC;Freire,Mila;Martinez,GerardoSanchez
- 通讯作者:Martinez,GerardoSanchez
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Marcelo Freire其他文献
Marcelo Freire的其他文献
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{{ truncateString('Marcelo Freire', 18)}}的其他基金
Exploring the Ecological Role of Rothia mucilaginosa and It's Iron Binding Siderophore Enterobactin
探索 Rothia mucilaginosa 及其铁结合铁载体肠杆菌素的生态作用
- 批准号:
10218502 - 财政年份:2021
- 资助金额:
$ 29.25万 - 项目类别:
Regulation of ChemR23 in Resolution of Inflammation
ChemR23 在炎症消退中的调节
- 批准号:
9750754 - 财政年份:2018
- 资助金额:
$ 29.25万 - 项目类别:
Regulation of ChemR23 in Resolution of Inflammation
ChemR23 在炎症消退中的调节
- 批准号:
8568302 - 财政年份:2013
- 资助金额:
$ 29.25万 - 项目类别:
Regulation of ChemR23 in Resolution of Inflammation
ChemR23 在炎症消退中的调节
- 批准号:
9076616 - 财政年份:2013
- 资助金额:
$ 29.25万 - 项目类别:
Regulation of ChemR23 in Resolution of Inflammation
ChemR23 在炎症消退中的调节
- 批准号:
8669965 - 财政年份:2013
- 资助金额:
$ 29.25万 - 项目类别: