Novel Determinants of Streptococcus Pyogenes Virulence and Protective Immunity in the Primate Oropharynx: A Genome-wide Strategy

灵长类口咽部化脓性链球菌毒力和保护性免疫的新决定因素：全基因组策略

• 批准号: 10387431
• 负责人: James MALLORY Musser
• 金额: $ 44万
• 依托单位: METHODIST HOSPITAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10387431
• 关键词: Acute; Address; Animal Model; Antibiotics; Biology; Candidate Disease Gene; Clinical; Complement; Consultations; Data; Disease; England; Epidemic; Equus caballus; Event; Female genitalia; Funding; Gene Deletion; Genes; Genomics; Goals; Gold; Grant; Human; Immunity; Immunization; Individual; Infection; Knowledge; Libraries; Macaca fascicularis; Membrane Proteins; Modeling; Molecular; Monobactams; Myositis; National Institute of Allergy and Infectious Disease; Necrotizing fasciitis; Organism; Oropharyngeal; Pathogenesis; Pharyngeal structure; Pharyngitis; Predisposition; Primates; Process; Proteins; Public Health; Publications; Recording of previous events; Research; Respiratory Tract Diseases; Role; Saliva; Septicemia; Serotyping; Severities; Site; Specimen; Streptococcus; Streptococcus equi; Streptococcus pyogenes; Study models; Technology; Testing; Upper Respiratory Infections; Upper respiratory tract; Vaccination; Vaccine Antigen; Vaccine Research; Vaccines; Virulence; Virulence Factors; Work; base; experience; fitness; gene product; genetic information; genome sequencing; genome wide screen; genome-wide; genome-wide analysis; human disease; improved; in vivo; innovation; mouse model; multiple myeloma M Protein; mutant; nonhuman primate; novel; novel vaccines; pathogen; pathogenic bacteria; prevent; public health relevance; reproductive tract; success; vaccine candidate; vaccine development; vaccinology

PROJECT SUMMARY The human bacterial pathogen Streptococcus pyogenes (group A streptococcus, GAS) causes more than 600 million cases of pharyngitis annually worldwide. Despite 100 years of effort, this organism has successfully defied attempts to create a vaccine that protects humans from pharyngitis and invasive infections. We believe a new strategy is warranted and necessary. The proposed research seeks to discover, on a genome-wide scale using a strategy not previously applied to GAS vaccinology, novel GAS genes contributing to pharyngitis, and exploit this information for vaccine research efforts. It is our thesis that this knowledge gap has severely limited the ability to fully understand the virulence determinants at work in the upper respiratory tract (URT) and create a protective GAS vaccine. We will address these issues by combining our recent successes applying transposon- directed insertion site sequencing (TraDIS) technology to GAS in several settings, together with our 19 years of productively using a cynomolgus macaque (nonhuman primate, NHP) model of pharyngitis, in vaccinology research. We will first use TraDIS to conduct in vivo genome-wide screens to systematically identify serotype M89 GAS genes required for colonization, clinical disease, and persistence in the URT of NHPs. These data will complement and enrich analogous information we have already generated for serotype M1 and M28 GAS. Our central hypothesis is that defining the GAS genes that contribute to fitness in the NHP URT will significantly improve our understanding of the molecular processes occurring in this niche, thereby filling a massive knowledge gap and leading to new strategies for creating a vaccine that protects against GAS pharyngitis. We propose the following three specific aims: Specific Aim 1: Exploit TraDIS for genome- wide identification of serotype M89 GAS genes required for colonization, acute clinical disease, and persistence (i.e., fitness) in the URT of NHPs. Specific Aim 2: Use our combined M1, M28, and M89 TraDIS URT screen data to generate isogenic gene-deletion mutant strains and validate the importance of additional selected candidate genes in causing GAS pharyngitis in NHPs. Specific Aim 3: Determine if vaccination of NHPs with proteins encoded by genes identified in Specific Aims 1 and 2 confers protective immunity against experimental pharyngitis caused by homologous (serotype M1) and heterologous (serotype M28) M protein serotypes of GAS. The proposed line of research will exploit and significantly expand studies funded by an R21 grant, and our innovative and successful application of TraDIS to GAS, our long history of GAS pharyngitis and vaccinology studies using NHPs, our analyses of human specimens, and our contributions to understanding the basic biology of GAS molecular pathogenesis and genomics. In essence, this research represents a new way forward for GAS vaccine efforts. Important to note, an analogous strategy has been used to identify four proteins present in a new vaccine that protects horses against “strangles,” a severe URT disease caused by the closely related pathogen, Streptococcus equi.

项目摘要 人类细菌病原体链球菌（A组链球菌，气体）导致超过600多个 全球每年有百万个咽炎病例。尽管付出了100年的努力，但这种有机体还是成功地反抗了 试图创建一种保护人类免受咽炎和侵入性感染的疫苗。我们相信一个新的 策略是必要的。拟议的研究旨在以全基因组规模发现 以前不适用于气体疫苗学，有助于咽炎的新型气体基因的策略和剥削 此信息用于疫苗研究工作。我们的论点是，这种知识差距严重限制了 能够充分理解病毒在上呼吸道（URT）中的工作中确定并创建一个 保护性气体疫苗。我们将通过结合使用转座子的成功来解决这些问题 将插入站点测序（Tradis）技术定向到多种环境中，以及我们的19年 在疫苗学中，有效地使用雌雄同体（非人类灵长类动物，NHP）模型 研究。我们将首先使用Tradis进行体内全基因组筛选以系统地识别血清型 NHP的URT中需要的M89气体基因才能定植，临床疾病和持久性。这些数据将 我们已经为血清型M1和M28气体生成的补体和丰富信息。我们的 中心假设是，定义有助于NHP URT适应性的气体基因将 显着提高了我们对在这个利基市场发生的分子过程的理解，从而 填补巨大的知识差距，并导致创建一种保护疫苗的新策略 气咽炎。我们提出以下三个特定目标：特定目标1：利用传统的基因组 - 广泛鉴定定植，急性临床疾病所需的血清型M89气体基因 （即健身）在NHP的URT中。特定目标2：使用我们组合的M1，M28和M89 Tradis URT屏幕 数据以生成等生基因脱落突变菌株并验证其他选定的重要性 候选基因引起NHP的气咽炎。特定目标3：确定NHP的疫苗接种是否与 在特定目的1和2中鉴定出的基因编码的蛋白质1和2供认保护性免疫针对实验 由同源性（血清型M1）和异源（血清型M28）M蛋白血清型引起的咽炎。 拟议的研究线将探索并大大扩大由R21赠款资助的研究，我们的 传统在天然气中的创新和成功应用，我们的天然气咽炎和疫苗学的悠久历史 使用NHP的研究，我们对人类标本的分析以及我们对理解基本生物学的贡献 气体分子发病机理和基因组学。本质上，这项研究代表了汽油的新方法 疫苗的努力。重要的是要注意，已经使用类似策略来识别存在的四种蛋白质 在一种保护马匹免受“绞死”的新疫苗中 相关病原体，链球菌当量。