A Rationally Targeted Approach to Preventing GBS Infection

预防 GBS 感染的合理针对性方法

• 批准号: 10076140
• 负责人: Thomas A Hooven
• 金额: $ 19万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-02-05 至 2022-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10076140
• 关键词: Amniotic Fluid; Animal Model; Antibodies; Antigens; Bacterial Infections; Basic Science; Binding; Bioinformatics; Biological; Blood; Data; Development; Disease; Essential Genes; Exposure to; Flow Cytometry; Genes; Genome; Goals; Growth; Health; Human; Immune; Immunization; Immunology; In Vitro; Infection; Interferon Type II; Interleukin-17; K-Series Research Career Programs; Knock-out; Knowledge; Laboratories; Libraries; Liquid substance; Mediating; Mentors; Mentorship; Methods; Modeling; Molecular Genetics; Mus; Neonatal; Neonatal Mortality; Oryctolagus cuniculus; Outcome; PAWR protein; Pathogenesis; Pediatrics; Perinatal Infection; Polysaccharides; Prevention; Prevention strategy; Proteins; Publishing; Research; Research Personnel; Scientific Advances and Accomplishments; Sepsis; Serotyping; Serum; Societies; Solid; Specificity; Streptococcus Group B; Surface; System; T cell response; T-Cell Activation; Techniques; Technology; Testing; Training; Universities; Vaccination; Vaccines; Vagina; Work; base; clinically relevant; early onset; efficacy testing; experimental study; fetal infection; fitness; gene product; genome wide screen; genome-wide; genomic locus; human disease; improved; in vivo; in vivo Model; infection risk; insight; intraamniotic infection; mortality; mouse model; mutant; neonatal morbidity; neonatal period; neonatal sepsis; next generation sequencing; novel; novel strategies; novel vaccines; pathogen; peripheral blood; polyclonal antibody; prevent; professor; programs; reproductive tract; screening; translational research program; transposon sequencing; vaccine candidate; vaccine development; vaccine efficacy; whole genome

PROJECT SUMMARY Dr. Thomas Hooven is a neonatologist and Assistant Professor of Pediatrics at Columbia University studying Streptococcus agalactiae (group B Streptococcus; GBS), the major infectious cause of neonatal morbidity and mortality. His research, under the mentorship of Dr. Adam Ratner, focuses on genes and gene networks that enable GBS to successfully colonize the maternal reproductive tract and to survive in amniotic fluid and blood during perinatal infection. Using a novel genome-wide screening technique based on next-generation sequencing of transposon-genome junctions from a saturated mutant library (Tn-seq), Dr. Hooven can accurately predict GBS genes whose protein products are necessary for bacterial growth under diverse experimental conditions. His proposed research seeks to use Tn-seq technology—in combination with ex vivo and in vivo models of colonization and invasion—to pinpoint surface-localized GBS proteins whose functions are essential for pathogenesis. Once validated by targeted knockout and antibody coincubation experiments, those proteins identified as essential for pathogenesis will be purified and tested as candidate vaccines to prevent vaginal colonization, ascending chorioamnionitis, and early-onset sepsis in clinically relevant mouse models. Recognizing that vaccine efficacy may depend on a combination of humoral and cellular immune mechanisms, this proposal also includes studies of opsonophagocytosis after antibody binding to candidate vaccine protein targets and T cell responses to immunization and vaginal colonization. By performing this research, Dr. Hooven will advance scientific understanding of GBS pathogenesis and bring society closer to a safe and effective vaccine to prevent devastating neonatal GBS infections. He will also expand his experimental repertoire through exposure to key methods in bioinformatics, immunology, molecular genetics, and vaccine development. This project will provide crucial training that will set the stage for his transition to becoming an independent investigator. Upon completion of the proposed research, Dr. Hooven will be ready to assume oversight of his own basic and translational research program aimed at advancing neonatal health through new insights into infection and new approaches toward its prevention and treatment.

项目概要 Thomas Hooven 博士是哥伦比亚大学的新生儿科医生和儿科助理教授 无乳链球菌（B 族链球菌；GBS）是新生儿发病的主要感染原因， 死亡。他的研究在 Adam Ratner 博士的指导下，重点关注基因和基因网络， 使 GBS 能够成功定植于母体生殖道并在羊水和血液中存活 围产期感染期间。使用基于下一代的新型全基因组筛选技术 Hooven 博士可以对饱和突变体文库中的转座子-基因组连接点进行测序 (Tn-seq) 准确预测 GBS 基因，其蛋白质产物是细菌在不同条件下生长所必需的 实验条件。他提出的研究旨在使用 Tn-seq 技术——结合离体 以及定植和入侵的体内模型——查明表面定位的 GBS 蛋白，其功能 对于发病机制至关重要。一旦通过靶向敲除和抗体共孵育实验验证， 那些被确定为发病机制所必需的蛋白质将被纯化并作为候选疫苗进行测试 预防临床相关小鼠的阴道定植、上行性绒毛膜羊膜炎和早发性败血症 模型。认识到疫苗功效可能取决于体液免疫和细胞免疫的结合 机制，该提案还包括抗体与候选物结合后调理吞噬作用的研究 疫苗蛋白靶标以及 T 细胞对免疫和阴道定植的反应。 通过进行这项研究，胡文博士将增进对 GBS 发病机制的科学认识，并带来 社会更接近安全有效的疫苗，以预防毁灭性的新生儿 GBS 感染。他还将 通过接触生物信息学、免疫学、分子学等领域的关键方法来扩展他的实验范围 遗传学和疫苗开发。该项目将提供重要的培训，为他的未来奠定基础 过渡为独立调查员。完成拟议的研究后，胡文博士将 准备好监督自己的基础和转化研究计划，旨在推进 通过对感染的新见解及其预防和治疗的新方法来促进新生儿健康。