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因果因素Borrelia Burgdorferi如何在自然界中生存 - 涉及研究其与天然哺乳动物储层宿主的关系， peromyscus leucopus。虽然先前的工作表明ixodes肩cap脚不成比例通过以无症状的白血病小鼠为食来获得B. burgdorferi（然后可以传递到人类）尚不清楚（a）为什么白疟原虫看起来比其他小啮齿动物更适合于B. burgdorferi，（b）为什么白链假单胞菌仍然不对称上皮慢性，全身性B. burgdorferi感染和（c）什么分子工具B. Burgdorferi用来成功感染白菜假单胞菌。阐明B. burgdorferi的自然生态代表一个重要的公共卫生目标，因为，B. burgdorferi感染通常可以治疗，但很重要几十年来，患者的少量一直是症状。那是找到防止B的机制。 Burgdorferi感染至关重要。该项目的长期目标是更好地了解共生 B. burgdorferi和P. leucopus之间的关系，以揭示破坏Enzootic循环的目标并减少莱姆病的燃烧。这项赠款旨在更好地理解白菜假单胞菌。 burgdorferi主机 - 微型互动（a）执行监测B. burgdorferi感染和白假单疟原虫感染的动态的实验性工作时间，（b）识别白疟原虫感染所需的B. burgdorferi基因。对于第一个目标，我会尝试了解细菌在白假单疟原虫组织中的燃烧如何与组织病理学，tick感染，随着时间的流逝，基因表达的变化。这将涉及对B进行最全面的研究。到目前在现场未经测试。为此，我将利用数字磁盘PCR，RNA测序，显微镜组织学分析和Xenodiagnostic测试。值得注意的是，我将简单地执行相同的实验两种杂交杂交线（C57BL/6J和C3H/HEN，它们会形成轻度和严重的莱姆病样因此，症状是将我的发现与经常使用的鼠模型进行比较。为了第二个目标将使用我的实验室经常在过去使用的突变库来部署转座测序筛选出版物。总而言之，这些实验将极大地改善我们对宿主和宿主的理解病原体因自然界生存的促进生存的病原体因子。此外，该项目非常适合促进我的长期目标是在数据集中在学术机构启动独立研究计划在这项研究中产生的将成为我未来实验室的基础。