Identifying Determinants of Borrelia burgdorferi and Peromyscus leucopus symbiosis

鉴定伯氏疏螺旋体和白白鼠共生的决定因素

• 批准号: 10751314
• 负责人: Jeffrey Bourgeois
• 金额: $ 6.91万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10751314
• 关键词: Acute; Affect; American; Bacteria; Bacterial Genes; Biological Assay; Black-legged Tick; Borrelia; Borrelia burgdorferi; CRISPR/Cas technology; Canis familiaris; Cell Line; Cells; Characteristics; Chronic; Data; Data Set; Defect; Development; Ecology; Elements; Enabling Factors; Experimental Designs; Fibroblasts; Foundations; Frequencies; Future; Gene Expression; Genes; Genetic; Genetic Screening; Glean; Goals; Grant; Health; Histology; Histopathology; House mice; Human; Immune response; Immunity; Immunologics; In Vitro; Inbreeding; Infection; Inflammation; Institution; Kinetics; Knock-out; Laboratories; Lead; Left; Libraries; Longitudinal Studies; Lyme Disease; Macrophage; Microscopic; Molecular; Monitor; Mus; Nature; Paper; Pathology; Pathway interactions; Patients; Peromyscus; Postdoctoral Fellow; Public Health; Publications; Publishing; Reagent; Research; Rodent; Rodent Model; Series; Side; Signal Pathway; Skin; Symbiosis; Symptoms; Techniques; Testing; Ticks; Time; Tissues; Training; United States; Work; cost; cytokine; data integration; digital; enzootic; experience; experimental study; feeding; follow-up; genome-wide; genomic tools; graduate school; health goals; host-microbe interactions; human pathogen; improved; in vivo; insight; interferon alpha receptor; mouse model; mutant; novel; pathogen; post-doctoral training; prevent; programs; response; screening; skills; tool; training opportunity; transcriptome sequencing; transmission process; transposon sequencing; vector tick

Project Summary/Abstract: My postdoctoral studies focus on understanding how Borrelia burgdorferi, the causal agent of Lyme disease, survives in nature—which involves studying its relationship with its natural mammalian reservoir host, Peromyscus leucopus. While previous work has demonstrated that Ixodes scapularis ticks disproportionately obtain B. burgdorferi (which can then be passed onto humans) by feeding on asymptomatic P. leucopus mice, it remains unclear (a) why P. leucopus appear better suited than other small rodents to pass on B. burgdorferi, (b) why P. leucopus remain asymptomatic despite chronic, systemic B. burgdorferi infection, and (c) what molecular tools B. burgdorferi uses to successfully infect P. leucopus. Elucidating the natural ecology of B. burgdorferi represents an important public health goal, as, while B. burgdorferi infection is typically treatable, a substantial fraction of patients remains symptomatic for months to decades. Thus, finding mechanisms to prevent B. burgdorferi infection is critical. The long-term goal of this project is to better understand the symbiotic relationship between B. burgdorferi and P. leucopus to reveal targets for disrupting the enzootic cycle and reducing the burden of Lyme disease. This grant seeks to better understand the P. leucopus-B. burgdorferi host-microbe interaction by (a) performing experimental work to monitor the dynamics of B. burgdorferi infection and P. leucopus inflammation change over time, and (b) identify B. burgdorferi genes that are required for P. leucopus infection. For the first goal, I will attempt to understand how bacterial burden in P. leucopus tissues correlates with histopathology, tick infectivity, and changes in gene expression over time. This will involve performing the most comprehensive study of B. burgdorferi infection in these rodents to date and will test a variety of hypotheses that have been postulated but left untested in the field. To accomplish this, I will leverage digital-droplet PCR, RNA-sequencing, microscopic histology analyses, and xenodiagnostic testing. Of note, I will simultaneously perform the same experiments in two Mus musculus inbred lines (C57BL/6J and C3H/HeN, which develop mild and severe Lyme disease-like symptoms, accordingly) to compare my findings against frequently used murine models. For the second aim, I will deploy transposon sequencing screening using a mutant library that my lab has frequently utilized in past publications. In sum, these experiments will dramatically improve our understanding of both the host and pathogen factors that facilitate B. burgdorferi survival in nature. Additionally, this project is well-suited to promote my long-term goal of starting an independent research program at an academic institution, as the datasets generated in this study will serve as the foundation of my future laboratory.

项目概要/摘要： 我的博士后研究集中在了解莱姆病的病原体伯氏疏螺旋体， 在自然界中生存-这涉及到研究它与其天然哺乳动物宿主的关系， 白足鹿鼠虽然以前的工作表明，肩突硬蜱不成比例地 得到B。通过喂养无症状的白足疟原虫小鼠， 目前尚不清楚（a）为什么白足鼠似乎比其他小型啮齿动物更适合传递B。伯氏菌，（B） 为什么尽管有慢性全身性B血症，白足疟原虫仍无症状。Burgdorferi感染，以及（c）什么分子 工具B。burgdorferi成功感染了P. leucopus。阐述了B的自然生态。burgdorferi 代表了一个重要的公共卫生目标，因为，而B.伯氏感染通常是可治疗的， 部分患者的症状持续数月至数十年。因此，找到预防B. 伯氏感染是致命的。这个项目的长期目标是更好地了解共生 B之间的关系。burgdorferi和P. leucopus揭示了破坏地方病循环的目标 减轻莱姆病的负担。 该资助旨在更好地了解白腹原杆菌-B。伯氏宿主-微生物相互作用，通过（a）进行 监测B动力学的实验工作。伯氏感染和白足巴斯德菌炎症转变 时间，以及（B）识别B。这些基因是白足巴斯德菌感染所需的伯氏螺旋体基因。对于第一个目标，我将 试图了解白足拟单胞菌组织中的细菌负荷如何与组织病理学，蜱感染性， 以及基因表达随时间的变化。这将涉及对B进行最全面的研究。 迄今为止，这些啮齿动物中的伯氏螺旋体感染，并将测试各种假设， 未经实地测试为了实现这一点，我将利用数字微滴PCR，RNA测序，显微镜， 组织学分析和异种诊断测试。值得注意的是，我将同时进行同样的实验， 两个小家鼠近交系（C57 BL/6 J和C3 H/HeN），它们具有轻度和重度莱姆病样病变， 症状），以将我的发现与常用的小鼠模型进行比较。第二个目标，我 我将使用我的实验室过去经常使用的突变体库来部署转座子测序筛选 出版物。总之，这些实验将大大提高我们对宿主和 促进B的病原体因素。burgdorferi在自然界中的生存。此外，该项目非常适合促进 我的长期目标是在一个学术机构开始一个独立的研究项目， 在这项研究中产生的将作为我未来实验室的基础。