Defining the impact of zinc on Streptococcus agalactiae biofilm and colonization

确定锌对无乳链球菌生物膜和定植的影响

• 批准号: 10358527
• 负责人: Ryan S Doster
• 金额: $ 2.87万
• 依托单位: VANDERBILT UNIVERSITY MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-09 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10358527
• 关键词: Academic Medical Centers; Adult; Area; Bacterial Infections; Bacterial Physiology; Cells; Clinical; Communicable Diseases; Complement; Core Facility; Data; Development; Epithelial; Epithelial Cells; Event; Female; Fetal Membranes; Future; Genetic; Genetic Transcription; Genomics; Glucose; Goals; Human; Immune response; Impairment; In Vitro; Infection; Inflammatory Response; Innate Immune Response; Light; Link; Medicine; Mentors; Mentorship; Microbial Biofilms; Microscopy; Modeling; Morbidity - disease rate; Mus; Mutagenesis; Newborn Infant; Nutrient; Pathogenesis; Pathway interactions; Pediatrics; Perinatal; Perinatal Infection; Physicians; Pregnancy; Pregnant Women; Premature Birth; Premature Labor; Production; Program Development; Regulator Genes; Regulatory Pathway; Research; Research Personnel; Resolution; Risk; Risk Factors; Role; Scholarship; Scientist; Stimulus; Streptococcal Infections; Streptococcus Group B; Supplementation; Techniques; Technology; Testing; Tissue membrane; Tissues; Training; United States; Vagina; Woman; Work; Zinc; Zinc deficiency; Zinc supplementation; career; career development; dietary; early onset; host colonization; human model; in vitro Model; in vivo; infection risk; innovation; instructor; instrument; intraamniotic infection; intrauterine infection; microbial host; mortality; mouse model; multidisciplinary; mutant; neonatal infection; neonatal sepsis; novel strategies; organ on a chip; pathogen; premature; prevent; reproductive tract; response; stillbirth; transcriptome; transcriptomics; zinc-binding protein

Project Summary During pregnancy, zinc deficiency is associated with an increased risk of infection. Streptococcus agalactiae, also known as Group B Streptococcus (GBS), colonizes 20-30% of pregnant women. GBS vaginal colonization increases the risk for stillbirth, preterm labor, and neonatal sepsis. One strategy that GBS may use to stably colonize the reproductive tract is to produce biofilm, and we have found that GBS biofilm production is suppressed in the presence of zinc. This proposal tests the hypothesis that GBS biofilm production is negatively regulated by zinc, and this biofilm contributes to stable colonization of host tissues by limiting innate immune responses against GBS. To test this hypothesis, Aim 1 will use mutagenesis and transcriptomic approaches to identify genetic pathways regulating GBS biofilm. Aim 2 will define the impact of zinc-regulated biofilm at the host-microbial interface by evaluating GBS mutants lacking cadD, a zinc efflux transporter, which results in impaired biofilm production. We will examine these interactions using in vitro models of human vaginal epithelial cells, explanted human fetal membranes, and an Instrumented Fetal Membrane Organ on a Chip (IFMOC) model under conditions that modify zinc availability. Aim 3 will use a mouse vaginal colonization model to define how the zinc transporter, cadD, impacts vaginal colonization in settings of zinc deficiency or supplementation. Completion of these Aims will shed new light on how zinc impacts GBS-host interactions that culminate in colonization (and infection) of the reproductive tract. Dr. Ryan Doster is an Instructor in the Division of Infectious Diseases at Vanderbilt University Medical Center. His past work investigated gestational tissue innate immune responses to infection. This project will expand his research focus to mechanisms of GBS colonization. This project complements Dr. Doster’s clinical training in pediatrics and adult infectious diseases medicine. Under the mentorship of Dr. David Aronoff, Dr. Jennifer Gaddy, and a multidisciplinary Scholarship Oversight Committee, Dr. Doster will master techniques including bacterial mutagenesis, transcriptomics, and modeling infection within the reproductive tract to determine how zinc alters bacterial physiology and pathogenesis. Vanderbilt offers cutting-edge technology and expertise in these areas through Vanderbilt Technologies for Advanced Genomics and other shared research core facilities. These studies and a strong career development program will prepare Dr. Doster for transition to an independent researcher investigating the pathogenesis of bacterial infections during pregnancy.

项目摘要 在怀孕期间，锌缺乏症与感染风险增加有关。链球菌agalactiae， 也称为B组链球菌（GBS），占20-30％的孕妇。 GBS阴道定殖 增加死产，早产劳动和新生儿败血症的风险。 GB可以使用的一种策略 殖民化生殖道是生产生物膜，我们发现GBS生物膜的产生是 在存在锌的情况下被抑制。该建议检验了GBS生物膜产生的假设是 受锌的负调节，这种生物膜通过限制先天而导致宿主组织的稳定定植 对GB的免疫反应。为了检验该假设，AIM 1将使用诱变和转录组学 鉴定调节GBS生物膜的遗传途径的方法。 AIM 2将定义锌调节的影响 通过评估缺少CADD的GBS突变体（锌外排运输蛋白）的GBS突变体，该突变体在宿主微生物界面处的生物膜（生物膜） 导致生物膜产生受损。我们将使用人的体外模型检查这些相互作用 阴道上皮细胞，外植的人类胎儿膜和仪器上的胎儿器官 在修改锌可用性的条件下，芯片（IFMOC）模型。 AIM 3将使用鼠标阴道定植 定义锌转运蛋白CADD如何影响阴道定植的模型 补充。这些目标的完成将为锌如何影响GBS-Host互动的新启示 在复制道的定植（和感染）中达到最终形式。 Ryan Doster博士是范德比尔特大学医学中心的传染病科讲师。 他过去的工作调查了妊娠组织先天免疫的感染。这个项目将扩大他的 研究重点是GBS定殖的机制。该项目完成了Doster博士的临床培训 儿科和成人传染病医学。在戴维·阿罗诺夫（David Aronoff）博士的心态下 Gaddy和多学科奖学金监督委员会，Doster博士将掌握包括 细菌诱变，转录组学和生殖道中的建模感染，以确定如何 锌改变细菌生理和发病机理。范德比尔特（Vanderbilt）提供了最先进的技术和专业知识 这些领域通过范德比尔特技术用于高级基因组学和其他共享研究核心 设施。这些研究和强大的职业发展计划将使Doster博士为过渡到 独立研究人员研究了怀孕期间细菌感染的发病机理。