Determinants of Streptococcus pyogenes fitness in the female primate genital tract: A genome-wide analysis

雌性灵长类生殖道中化脓性链球菌适应性的决定因素：全基因组分析

• 批准号: 9805040
• 负责人: James MALLORY Musser
• 金额: $ 20.19万
• 依托单位: METHODIST HOSPITAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-06-01 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9805040
• 关键词: Address; Anatomy; Animals; Bacterial Genes; Basic Science; Binding; Biology; Blood; Candidate Disease Gene; Clinical; Complement; Data; Diagnosis; Disease; Environment; Epidemic; Epithelium; Event; Female; Future; Gene Silencing; Genes; Genome; Goals; Gold; Human; In Vitro; Individual; Infection; Knowledge; Libraries; Macaca; Medicine; Methods; Modeling; Molecular; Monitor; Morbidity - disease rate; Motivation; Mouse Strains; Mucous Membrane; Mutagenesis; Myositis; Necrotizing fasciitis; Neonatal; Organism; Pathogenesis; Pharyngitis; Phase; Phylogenetic Analysis; Primates; Process; Puerperal Infection; Puerperal pyrexia; Recording of previous events; Reporting; Research; Research Activity; Role; Saliva; Sepsis; Septicemia; Serotyping; Severities; Site; Streptococcus pyogenes; Surface; Targeted Research; Technology; Time; Translational Research; Vaccines; Vagina; Virulence; Virulence Factors; Work; base; design; experience; fitness; gene product; genome sequencing; genome wide screen; genome-wide; genome-wide analysis; human disease; human pathogen; in vivo; innovation; interest; knockout gene; mortality; mouse model; mucosal site; mutant; neonatal sepsis; nonhuman primate; novel; pathogen; pathogenic bacteria; prevent; reproductive tract; subcutaneous; success; tool; transcriptome; transcriptome sequencing; translational approach; vaccine candidate

PROJECT SUMMARY The human bacterial pathogen Streptococcus pyogenes (group A streptococcus, GAS) causes more than 700 million human infections annually worldwide. The proposed research seeks to discover, on a genome- wide scale, GAS genes required for, or contributing to fitness in the female genital tract, an infection site for which very little basic understanding is known. We will use a serotype M28 GAS strain because for reasons that aren’t understood, these organisms are non-randomly associated with female genital tract disease, including puerperal sepsis (childbed fever) and neonatal invasive infections. The pathogen genes and fundamental molecular pathogenesis processes contributing to two phases of genital tract interaction will be studied: (i) colonization and proliferation and (ii) persistence. This basic science knowledge deficit has severely limited the ability to create new clinical tools such as a successful GAS vaccine. Thus, the studies are designed to address this critical knowledge deficit. We will combine our very recent success in applying transposon-directed insertion site sequencing (TraDIS) technology to GAS with our 18 years of productive experience with cynomolgus macaque models of GAS infection, currently the gold standard experimental animal to study this human pathogen. We will use TraDIS to conduct an in vivo genome-wide screen that will systematically identify GAS genes required for colonization and proliferation, and persistence in the genital tract of female cynomolgus macaques. The goal of the proposed studies is to identify new molecular mechanisms used by GAS to enhance fitness in this anatomic niche. The results of our studies may ultimately help in formulating novel translational strategies to prevent or treat GAS infections. To achieve our goals, the following two specific aims are proposed: Specific Aim 1: Exploit TraDIS for genome-wide identification of GAS genes required for (i) colonization and proliferation, and (ii) persistence (i.e., fitness) in the genital tract of female cynomolgus macaques. Specific Aim 2: Use isogenic gene-knockout mutant strains to validate the importance of four selected candidate genes to GAS colonization and proliferation, and persistence identified by the genome-wide screen. The proposed line of research will exploit our innovative and successful application of TraDIS to GAS, including infections in cynomolgus macaques. It will be the first use of TraDIS in the genital tract of female non-human primates and the first study to investigate determinants of GAS colonization and proliferation, and persistence, on a mucosal surface using genome-wide transposon mutagenesis.

项目概要 人类细菌病原体化脓性链球菌（A 族链球菌，GAS）可引起超过 全球每年有 7 亿人感染。拟议的研究旨在发现，在基因组上 广泛的GAS基因需要或有助于女性生殖道的健康，女性生殖道是感染部位 人们对此知之甚少。我们将使用血清型 M28 GAS 菌株，因为以下原因 目前尚不清楚，这些生物体与女性生殖道疾病非随机相关， 包括产后败血症（产褥热）和新生儿侵袭性感染。病原体基因和 促成生殖道相互作用的两个阶段的基本分子发病机制将是 研究了：（i）定植和增殖以及（ii）持久性。这种基础科学知识的匮乏 严重限制了创造新的临床工具（例如成功的 GAS 疫苗）的能力。因此，研究 旨在解决这一关键的知识缺陷。我们将结合我们最近在申请方面取得的成功 凭借我们 18 年的丰富经验，将转座子定向插入位点测序 (TraDIS) 技术应用于 GAS 食蟹猴GAS感染模型的经验，目前金标准实验 动物来研究这种人类病原体。我们将使用 TraDIS 进行体内全基因组筛选， 系统地鉴定定殖和增殖以及在生殖器中持续存在所需的 GAS 基因 雌性食蟹猴的肠道。拟议研究的目标是确定新的分子 GAS 使用的机制来增强该解剖学生态位的适应性。我们的研究结果可能最终 帮助制定新的转化策略来预防或治疗 GAS 感染。为了实现我们的目标， 提出了以下两个具体目标： 具体目标 1：利用 TraDIS 进行全基因组鉴定 (i) 定植和增殖以及 (ii) 在生殖道中持续存在（即适应性）所需的 GAS 基因 雌性食蟹猴。具体目标2：利用等基因基因敲除突变株来验证 四个选定的候选基因对 GAS 定植和增殖以及持久性的重要性已确定 通过全基因组筛选。拟议的研究路线将利用我们的创新和成功 TraDIS 在 GAS 中的应用，包括食蟹猴感染。这将是 TraDIS 的首次使用 在雌性非人类灵长类动物的生殖道中进行的研究以及第一项调查 GAS 决定因素的研究 使用全基因组转座子在粘膜表面定植、增殖和持久性 诱变。