In vivo dual Bioluminescence Reporter System of Infectious Borrelia burgdorferi
传染性伯氏疏螺旋体的体内双生物发光报告系统
基本信息
- 批准号:8497620
- 负责人:
- 金额:$ 20.53万
- 依托单位:
- 依托单位国家:美国
- 项目类别:
- 财政年份:2012
- 资助国家:美国
- 起止时间:2012-07-01 至 2016-06-30
- 项目状态:已结题
- 来源:
- 关键词:AddressAmericanAntibioticsAntigensArthritisArthropod VectorsArthropodsBacteria sigma factor KatF proteinBioluminescenceBorreliaBorrelia burgdorferiBorrelia oxidative stress regulatorCarbon DioxideCardiacCase StudyCenters for Disease Control and Prevention (U.S.)CharacteristicsChronicCodon NucleotidesConnective TissueCuesDataDiagnosisDiseaseEnvironmentEvaluationExanthemaGene ExpressionGene Expression ProfileGene TargetingGenesGenetic TranscriptionGoalsImageIncidenceInfectionInfectious AgentKineticsLaboratoriesLifeLigand Binding DomainLightLipoproteinsLuciferasesLyme DiseaseModalityModelingMonitorMusNeedlesNeurologicNorth AmericaOrder SpirochaetalesOrganismOspC proteinOxygenPathogenesisPathologyPatientsPatternPhasePlayProcessProductionProteinsPublic HealthPublishingRegulationRegulatory PathwayReporterReportingRoleSkinStagingStudy SectionSurfaceSystemTechniquesTechnologyTemperatureTestingTick-Borne DiseasesTicksTimeTissue-Specific Gene ExpressionTranscriptional ActivationUnited StatesVirulenceVisionWorkbasedesignerythema migransfluin vivoinnovationinsightmemberpathogenpromoterresponsesuccesstissue tropismtransmission processvector
项目摘要
DESCRIPTION (provided by applicant): It is well established that Borrelia burgdorferi, the etiologic agent of Lyme disease, modulates gene expression during infection as it moves between an arthropod vector and mammalian hosts. Several genes required for the establishment of mammalian infection are known with the prototypical gene being ospC. The lipoprotein OspC is absolutely required for mammalian infectivity, and although a putative ligand-binding domain is essential for infectivity, the exact function of OspC is not known. Subsequent studies indicated that ospC is coordinately regulated via a response regulator (Rrp2) that, together with RpoN, drives the expression of RpoS, which then promotes the transcriptional activation of ospC and other infectivity-associated borrelial genes. However, the activation of ospC is transient as it is repressed following infection. In this regard, if ospC expression is made constitutive, the spirochetes are rapidly cleared. Despite this observation, the kinetics of ospC expression, i.e., the amplitude and diminution within a living system over time, is not known.
Recently we have used in vivo imaging to detect light emitting (i.e., luciferase [luc] expressing) infectious B. burgdorferi following needle inoculation in mice. The advantage of this approach is that B. burgdorferi can be visualized numerous times in live mice over time to track the infectious process. Given the sensitivity of this technique, an additional potential applicatio might be to assess the expression of targeted genes. To test this premise, we have fused the ospC promoter (PospC) to luc. Our Preliminary Data suggests that ospC is highly expressed early in the infectious process within skin, but is significantly reduced later in the infection, consistent with prior reports indicating that it is down regulated following colonization and dissemination. The utility of this approach will now be expanded to further study the spatial expression or ospC as well as other genes that are coordinately regulated with ospC via RpoS. To this end we propose the following Specific Aims: (1) Characterize the in vivo tissue tropism and temporal production of borrelial ospC utilizing a dual bioluminescence reporter system; and (2) Determine the in vivo expression patterns of genes involved in the Rrp2-RpoN-RpoS regulatory pathway. In the proposed studies, the fate of ospC transcription will be tracked following disseminated infection as well as genes involved in the infectious process to determine if RpoS regulation is highly coordinated or occurs at differential times as B. burgdorferi disseminates.
The ability to visualize these regulatory patterns of specific borrelial promoter-luc constructs, focusing on the promoters of genes known to be involved in experimental mouse infection, should provide important insight into the hierarchy and/or temporal expression of these loci to establish and maintain an infectious focus. This exciting approach provides a powerful non-invasive, real time modality to evaluate the activity of a given promoter in a temporal and spatial manner.
描述(由申请人提供):众所周知,莱姆病的病因学伯氏伯氏菌(Borrelia Burgdorferi)在感染过程中调节基因表达,因为它在节肢动物载体和哺乳动物宿主之间移动。建立哺乳动物感染所需的几种基因已知原型基因是OSPC。脂蛋白OSPC绝对需要哺乳动物感染性,尽管假定的配体结合结构域对于感染性至关重要,但OSPC的确切功能尚不清楚。随后的研究表明,OSPC通过响应调节剂(RRP2)协调调节,该响应调节剂与RPON一起驱动RPOS的表达,然后促进OSPC和其他与感染相关的杂货基因的转录激活。但是,OSPC的激活是短暂的,因为它在感染后被抑制。在这方面,如果将OSPC表达为组成型,则将迅速清除螺旋体。尽管有这样的观察,但尚不清楚OSPC表达的动力学,即随着时间的流逝,活体系中的幅度和减小。
最近,我们已经使用体内成像来检测小鼠接种针头接种后的发光(即表达荧光素酶[Luc]表达)感染性B. burgdorferi。这种方法的优点是,随着时间的推移,可以在活小鼠中多次将B. burgdorferi可视化,以跟踪传染性过程。鉴于该技术的敏感性,可能是评估靶向基因的表达的其他潜在应用。为了测试这个前提,我们将OSPC启动子(POSPC)融合到LUC。我们的初步数据表明,OSPC在皮肤内感染过程的早期高度表达,但在感染后期显着降低,这与先前的报告一致,表明在定植和传播后,它被降低了。现在将扩展该方法的实用性,以进一步研究空间表达或OSPC以及其他通过RPOS协调调节的基因。为此,我们提出了以下特定目的:(1)表征利用双重发光报道器系统的体内组织的向量和颞骨OSPC的时间产生; (2)确定参与RRP2-RPON-RPOS调节途径的基因的体内表达模式。在拟议的研究中,将在传播感染后跟踪OSPC转录的命运以及传染过程中涉及的基因,以确定RPOS调节是否高度协调或在差异时间发生为Burgdorferi B. burgdorferi传播。
可视化特定杂质启动子 - 肺活构建体的这些调节模式的能力,重点是已知参与实验小鼠感染的基因启动子,应提供对这些基因座的层次结构和/或时间表达的重要见解,以建立和保持感染力。这种令人兴奋的方法提供了一种强大的非侵入性实时方式,可以以时间和空间方式评估给定启动子的活动。
项目成果
期刊论文数量(3)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
In Vivo Imaging Demonstrates That Borrelia burgdorferi ospC Is Uniquely Expressed Temporally and Spatially throughout Experimental Infection.
体内成像表明伯氏疏螺旋体 ospC 在整个实验感染过程中在时间和空间上都有独特的表达。
- DOI:10.1371/journal.pone.0162501
- 发表时间:2016
- 期刊:
- 影响因子:3.7
- 作者:Skare,JonathanT;Shaw,DanaK;Trzeciakowski,JeromeP;Hyde,JennyA
- 通讯作者:Hyde,JennyA
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Jenny A. Hyde其他文献
Jenny A. Hyde的其他文献
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{{ truncateString('Jenny A. Hyde', 18)}}的其他基金
Regulatory Pathways in Borrelial Pathogenesis
伯氏疏螺旋体发病机制的调节途径
- 批准号:
10677726 - 财政年份:2022
- 资助金额:
$ 20.53万 - 项目类别:
Regulatory Pathways in Borrelial Pathogenesis
伯氏疏螺旋体发病机制的调节途径
- 批准号:
10504708 - 财政年份:2022
- 资助金额:
$ 20.53万 - 项目类别:
Elucidating the Bacterial and Host Mechanisms Governing B. burgdorferi-Related Type I Interferon Responses
阐明控制伯氏疏螺旋体相关 I 型干扰素反应的细菌和宿主机制
- 批准号:
10302429 - 财政年份:2021
- 资助金额:
$ 20.53万 - 项目类别:
The role of small non-coding RNA in borrelial pathogenesis
小非编码 RNA 在疏螺旋体发病机制中的作用
- 批准号:
9090929 - 财政年份:2016
- 资助金额:
$ 20.53万 - 项目类别:
Characterization of C02 Sensing and Regulatory Response in Borrelia burgdorferi
伯氏疏螺旋体中CO2感应和调节反应的表征
- 批准号:
8715687 - 财政年份:2013
- 资助金额:
$ 20.53万 - 项目类别:
Characterization of C02 Sensing and Regulatory Response in Borrelia burgdorferi
伯氏疏螺旋体中CO2感应和调节反应的表征
- 批准号:
8583139 - 财政年份:2013
- 资助金额:
$ 20.53万 - 项目类别:
In vivo dual Bioluminescence Reporter System of Infectious Borrelia burgdorferi
传染性伯氏疏螺旋体的体内双生物发光报告系统
- 批准号:
8358909 - 财政年份:2012
- 资助金额:
$ 20.53万 - 项目类别:
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