A Rationally Targeted Approach to Preventing GBS Infection

项目总结 托马斯·霍芬博士是一名新生儿科医生，同时也是哥伦比亚大学儿科学的助理教授。 无乳链球菌(B群链球菌；GBS)，新生儿发病率和 死亡率。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能够成功地在产妇生殖道中定居，并在羊水和血液中存活 在围产期感染期间。使用基于下一代的新的全基因组筛选技术 来自饱和突变文库的转座子-基因组连接的测序(TN-seq)，胡芬博士可以 准确预测细菌在不同环境下生长所必需的GBS基因 实验条件。他提出的研究试图将TN-SEQ技术与体外实验相结合 以及体内的定植和侵袭模型--精确定位表面定位的GBS蛋白，其功能 在发病机制中是必不可少的。一旦通过靶向基因敲除和抗体共孵育实验进行验证， 那些被确认为致病必需的蛋白质将被提纯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 模特们。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，这项建议还包括抗体与候选抗体结合后的吞噬细胞作用的研究 疫苗蛋白靶点和T细胞对免疫和阴道定植的反应。 通过进行这项研究，霍芬博士将促进对GBS发病机制的科学理解，并为 社会更接近安全有效的疫苗，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、分子生物学的关键方法来扩展他的实验曲目 遗传学和疫苗研发。该项目将提供关键的培训，为他的 过渡成为一名独立调查员。在完成拟议的研究后，胡芬博士将 准备好监督他自己的基础和翻译研究计划，旨在推动 通过对感染的新见解和预防和治疗的新方法，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.