A Rationally Targeted Approach to Preventing GBS Infection

项目摘要 博士托马斯霍芬是哥伦比亚大学的儿科学助理教授， 无乳链球菌（B组链球菌; GBS）是新生儿发病的主要感染性原因， mortality.他的研究，在亚当·拉特纳博士的指导下，专注于基因和基因网络， 使GBS能够成功地定殖于母体生殖道并在羊水和血液中存活 围产期感染。使用一种新的全基因组筛选技术， Hooven博士从一个饱和的突变体库（Tn-seq）中测序转座子-基因组连接， 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-结合体外 和体内模型的殖民和入侵，以查明表面定位的GBS蛋白，其功能 对发病机理至关重要一旦通过靶向敲除和抗体共孵育实验验证， 那些被鉴定为致病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、上行性绒毛膜炎和早发性脓毒症 模型认识到疫苗的效力可能取决于体液免疫和细胞免疫的结合， 机制，该提案还包括抗体结合候选物后调理吞噬作用的研究。 疫苗蛋白靶点和T细胞对免疫和阴道定殖的应答。 通过进行这项研究，Hooven博士将推进对GBS发病机制的科学认识， 社会更接近一个安全有效的疫苗，以防止毁灭性的新生儿GBS感染。他还将 通过接触生物信息学、免疫学、分子生物学和生物医学领域的关键方法， 遗传学和疫苗开发。这个项目将提供关键的培训，将为他奠定基础。 成为独立调查员。在完成拟议的研究后，何芬博士将 准备好承担他自己的基础和转化研究计划的监督，旨在推进 通过对感染的新认识以及预防和治疗感染的新方法，促进新生儿健康。

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.