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 人类细菌病原体化脓性链球菌（A组链球菌，GAS）导致超过600 全世界每年有100万例咽炎。尽管经过了100年的努力，这种生物成功地 试图制造一种疫苗，保护人类免受咽炎和侵入性感染。我们相信一个新的 战略是必要的和必要的。这项拟议中的研究试图在全基因组范围内发现， 一种以前没有应用于GAS疫苗学的策略，新的GAS基因有助于咽炎，并利用 这些信息用于疫苗研究。我们的论点是，这种知识差距严重限制了 能够充分了解上呼吸道（URT）中的毒力决定因素，并创建一个 保护性GAS疫苗我们将结合我们最近成功应用转座子- 定向插入位点测序（TraDIS）技术在几种情况下用于GAS，以及我们19年来 在疫苗学中有效地使用食蟹猴（非人灵长类动物，NHP）咽炎模型 research.我们将首先使用TraDIS进行体内全基因组筛选，以系统地鉴定血清型 NHP定植、临床疾病和在URT中的持久性所需的M89 GAS基因。这些数据将 补充和丰富了我们已经为血清型M1和M28 GAS生成的类似信息。我们 中心假设是，定义有助于NHP URT适应性的GAS基因将 显着提高我们的理解分子过程中发生的这个利基，从而 填补了巨大的知识空白，并导致新的战略，创造一种疫苗， GAS咽炎。我们提出了以下三个具体目标：具体目标1：利用TraDIS进行基因组- 广泛鉴定定植、急性临床疾病和持久性所需的血清型M89 GAS基因 (i.e.,在NHP的URT中。具体目标2：使用我们的组合M1、M28和M89 TraDIS URT屏幕 数据，以产生同基因基因缺失突变株，并验证额外选择的重要性， 在NHP中引起GAS咽炎的候选基因。具体目标3：确定NHP是否接种 由特异性目的1和2中鉴定的基因编码的蛋白质赋予针对实验性免疫的保护性免疫力。 由GAS的同源（血清型M1）和异源（血清型M28）M蛋白血清型引起的咽炎。 拟议的研究路线将利用并大大扩展由R21赠款资助的研究， 创新和成功地将TraDIS应用于GAS，我们在GAS咽炎和疫苗学方面的悠久历史 使用NHP的研究，我们对人类标本的分析，以及我们对理解基础生物学的贡献 GAS分子发病机制和基因组学的研究。从本质上讲，这项研究代表了GAS的一条新的前进道路。 疫苗的努力。值得注意的是，一个类似的策略已被用于确定四个蛋白质存在 在一种新的疫苗，保护马对“扼杀”，一个严重的URT疾病引起的密切 相关病原体马链球菌