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

基本信息

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
项目类别：
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

项目摘要

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.

项目摘要/摘要：我的博士后研究重点是了解莱姆病的病原体伯氏疏螺旋体如何在自然界中生存——这涉及研究它与其天然哺乳动物储存宿主的关系，虽然之前的研究表明肩胛硬蜱的蜱虫比例不成比例。通过喂养无症状的白种伯氏杆菌小鼠获得伯氏疏螺旋体（然后可以传染给人类），它仍不清楚 (a) 为什么白斑白腹鼠比其他小型啮齿动物更适合传播伯氏疏螺旋体，(b) 为什么白种伯氏杆菌在慢性、全身性伯氏疏螺旋体感染的情况下仍然无症状，以及 (c) 是什么分子伯氏疏螺旋体用来成功感染白叶伯氏菌的工具阐明伯氏疏螺旋体的自然生态。代表了一个重要的公共卫生目标，因为虽然伯氏疏螺旋体感染通常是可以治疗的，但大量的一小部分患者的症状持续数月至数十年，因此，寻找预防伯氏杆菌的机制。该项目的长期目标是更好地了解伯氏杆菌感染的共生性。伯氏疏螺旋体和白拟杆菌之间的关系揭示了破坏地方性动物循环的目标并减轻莱姆病的负担。该资助旨在通过 (a) 执行更好地了解白种伯氏杆菌 (P. burgdorferi) 宿主与微生物的相互作用。监测伯氏疏螺旋体感染动态和白色伯氏杆菌炎症变化的实验工作时间，以及 (b) 识别伯氏疏螺旋体感染所需的基因对于第一个目标，我将。尝试了解 P. leucopus 组织中的细菌负荷与组织病理学、蜱感染性、以及基因表达随时间的变化，这将涉及对 B. 迄今为止，这些啮齿动物感染了伯氏杆菌，并将检验各种已提出的假设，但为了实现这一目标，我将利用数字微滴 PCR、RNA 测序、显微镜。值得注意的是，我将在组织学分析和异种诊断测试中同时进行相同的实验。两个小家鼠近交系（C57BL/6J 和 C3H/HeN，它们会发展出轻度和重度莱姆病样症状）相应的症状）将我的发现与常用的小鼠模型进行比较对于第二个目标，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总之，这些实验将极大地提高我们对宿主和宿主的理解。此外，该项目非常适合促进伯氏疏螺旋体在自然界中生存的病原体因素。我的长期目标是在学术机构启动独立研究项目，如数据集这项研究中产生的成果将作为我未来实验室的基础。