Virulence Determinants of Borrelia burgdorferi

伯氏疏螺旋体的毒力决定因素

• 批准号: 9102864
• 负责人: STEVEN J. NORRIS
• 金额: $ 43.08万
• 依托单位: UNIVERSITY OF TEXAS HLTH SCI CTR HOUSTON
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9102864
• 关键词: Adherence; Affect; Arthritis; Arthropods; Bacteria; Biological Assay; Black-legged Tick; Blood capillaries; Borrelia; Borrelia burgdorferi; C3H/HeN Mouse; Cardiovascular system; Chemotaxis; Communities; Data; Dermal; Development; Diagnosis; Disease; Evaluation; Feeding Methods; Gene Expression Regulation; Genes; Genetic study; Genome; Goals; Grant; Health; Heart; Human; Immersion Investigative Technique; Infection; Inflammatory Response; Invaded; Ixodes; Joints; Libraries; Lyme Disease; Mammals; Membrane Proteins; Methodology; Methods; Mus; Mutagenesis; Mutate; Mutation; Nervous system structure; Neurologic; North America; Nutrient; Order Spirochaetales; Organism; Parents; Pathogenesis; Pathway interactions; Phase; Plasmids; Prevention; Process; Proteins; Protocols documentation; Replicon; Research; Research Personnel; Resource Sharing; Resources; Role; Sampling; Screening procedure; Shapes; Site; Site-Directed Mutagenesis; Skin; Specimen; Symptoms; Techniques; Testing; Tick Infestations; Ticks; Time; Tissues; Toxin; United States; Virulence; base; capillary; cell motility; design; feeding; gene complementation; improved; interest; mutant; novel; pathogen; prevent; screening; transmission process

DESCRIPTION (provided by applicant): Borrelia burgdorferi is the causative agent of Lyme disease in North America and is transmitted by ticks of the genus Ixodes. It is a highly invasive spirochete that can cause infection and manifestations in humans and other mammals that persist for months to years. The infection has localized, disseminated, and persistent phases, and B. burgdorferi appears to cause dermal, neurologic, cardiovascular, and arthritic symptoms primarily though the ability to invade almost any tissue, establish long-term infection, and induce inflammatory responses. The bacterium produces no known toxins, and its mechanisms of pathogenesis are largely unknown. Genetic studies using low-passage, infectious B. burgdorferi have been challenging due to exceedingly low transformation rates and plasmid loss; as a result, fewer than 50 genes have been investigated by allelic exchange or other site-directed mutagenesis methods for their importance in the mammal-tick infectious cycle. During the prior grant period, a sequence-defined library of 4,479 signature-tagged mutagenesis (STM) transposon mutants was generated in a transformable, infectious clone of B. burgdorferi B31. The plasmid content of the clones in this library was also determined using a novel Luminex-based high throughput strategy. Using the library, we have already performed STM screening of mouse infectivity of 484 transposon mutant clones in 434 different genes. In Aim 1, we will complete the systematic analysis of the roles of the 790 mutated protein-encoding genes in the infection of C3H/HeN mice using the STM mutant library. For this analysis, we will employ the STM Luminex Assay protocol developed during the prior grant period to analyze each mutant clone in 3 mice per group, 5 different tissues, and two time points in a high throughput manner. The goal of Aim 2 is to determine the effects of the transposon mutations on infectivity, persistence and transmission of B. burgdorferi in Ixodes scapularis ticks. Groups of the STM mutants will be transmitted to mice by larval ticks infected by immersion with a mixture of organisms with different mutations and signature tags. The survival of the STM clones will be analyzed before and after feeding on mice, and transmission of the clones to the mice will also be determined by the STM Luminex Assay. In Aim 3, the findings in Aims 1 and 2 will be used to select classes of mutants for detailed infectivity, complementation, and functional analysis. These analyses will include studies of genes involved in chemotaxis, motility, nutrient transport, surface proteins involved in adherence and other roles, novel gene regulation pathways, and additional gene sets identified during the STM screening procedure. The STM library will also be made available to any interested investigator through BEI-Resources, providing maximal resource sharing and stimulation of Borrelia research. We anticipate that this project will continue to fuel new discoveries useful in improving the diagnosis, treatment, and prevention of Lyme borreliosis.

描述（由申请人提供）：伯氏疏螺旋体是北美莱姆病的病原体，通过硬蜱属蜱传播。它是一种高度侵入性的螺旋体，可引起人类和其他哺乳动物的感染和症状，并持续数月至数年。感染具有局部、播散和持续阶段，伯氏疏螺旋体似乎主要通过侵入几乎任何组织、建立长期感染并诱发皮肤、神经、心血管和关节炎症状。 炎症反应。该细菌不产生已知的毒素，其发病机制也很大程度上未知。由于转化率极低和质粒丢失，使用低传代、传染性伯氏疏螺旋体进行遗传研究一直具有挑战性；因此，通过等位基因交换或其他定点诱变方法研究了不到 50 个基因在哺乳动物蜱感染周期中的重要性。在之前的资助期间，在伯氏疏螺旋体 B31 的可转化、感染性克隆中生成了包含 4,479 个特征标签诱变 (STM) 转座子突变体的序列定义文库。还使用基于 Luminex 的新型高通量策略测定了该文库中克隆的质粒含量。使用该文库，我们已经对 434 个不同基因的 484 个转座子突变克隆的小鼠感染性进行了 STM 筛选。在目标1中，我们将利用STM突变体库完成对790个突变蛋白编码基因在C3H/HeN小鼠感染中的作用的系统分析。对于此分析，我们将采用在先前资助期间开发的 STM Luminex 检测方案以高通量方式分析每组 3 只小鼠、5 个不同组织和两个时间点的每个突变克隆。目标 2 的目标是确定转座子突变对肩胛硬蜱中伯氏疏螺旋体的感染性、持久性和传播的影响。成群的 STM 突变体将通过幼虫蜱传播给小鼠，幼虫蜱通过与具有不同突变和特征标签的生物体混合物浸泡而感染。将在喂食小鼠之前和之后分析 STM 克隆的存活情况，并且还将通过 STM Luminex Assay 确定克隆向小鼠的传播情况。在目标 3 中，目标 1 和 2 中的发现将用于选择突变体类别，以进行详细的感染性、互补性和功能分析。这些分析将包括对参与趋化性、运动性、营养物质运输的基因、参与粘附和其他作用的表面蛋白、新的基因调控途径以及在 STM 筛选过程中识别的其他基因组的研究。 STM 图书馆还将通过 BEI-Resources 向任何感兴趣的研究人员开放，从而提供最大限度的资源共享和伯氏疏螺旋体研究的刺激。我们预计该项目将继续推动新发现，有助于改善莱姆疏螺旋体病的诊断、治疗和预防。

项目成果

Peaceful coexistence amongst Borrelia plasmids: Getting by with a little help from their friends?

• DOI: 10.1016/j.plasmid.2013.05.002
• 发表时间: 2013-09-01
• 期刊: PLASMID
• 影响因子: 2.6
• 作者: Chaconas, George;Norris, Steven J.
• 通讯作者: Norris, Steven J.

The dynamic proteome of Lyme disease Borrelia.

莱姆病疏螺旋体的动态蛋白质组。

• DOI: 10.1186/gb-2006-7-3-209
• 发表时间: 2006
• 期刊: Genome biology
• 影响因子: 12.3
• 作者: Norris,StevenJ

Strict Conservation yet Non-Essential Nature of Plasmid Gene bba40 in the Lyme Disease Spirochete Borrelia burgdorferi.

莱姆病螺旋体伯氏疏螺旋体中质粒基因 bba40 的严格保守性但非必需性质。

• DOI: 10.1128/spectrum.00477-23
• 发表时间: 2023-06-15
• 期刊: Microbiology spectrum
• 影响因子: 3.7

Analysis of an ordered, comprehensive STM mutant library in infectious Borrelia burgdorferi: insights into the genes required for mouse infectivity.

• DOI: 10.1371/journal.pone.0047532
• 发表时间: 2012
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Lin T;Gao L;Zhang C;Odeh E;Jacobs MB;Coutte L;Chaconas G;Philipp MT;Norris SJ
• 通讯作者: Norris SJ

A high-throughput genetic screen identifies previously uncharacterized Borrelia burgdorferi genes important for resistance against reactive oxygen and nitrogen species.

• DOI: 10.1371/journal.ppat.1006225
• 发表时间: 2017-02
• 期刊: PLoS pathogens
• 影响因子: 6.7
• 作者: Ramsey ME;Hyde JA;Medina-Perez DN;Lin T;Gao L;Lundt ME;Li X;Norris SJ;Skare JT;Hu LT
• 通讯作者: Hu LT