Administrative Diversity Supplement for Aimee Potter

艾米·波特的行政多元化补充

基本信息

批准号：
10818164
负责人：
CYNTHIA N CORNELISSEN
金额：
$ 13.7万
依托单位：
GEORGIA STATE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-08-09 至 2027-07-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10818164
关键词：
Affect Affinity Algorithms Animals Antibiotic Resistance Antibiotics Award Bacteria Binding Binding Proteins Biochemical Biochemical Pathway Biological Assay Biological Availability Biology Centers for Disease Control and Prevention (U.S.)Chemicals Complement Complex Data Data Set Dependence Energy Supply Enterobactin Environment Enzyme-Linked Immunosorbent Assay Epithelium Erythrocytes Exposure to Future Genes Genetic Genetic Transcription Goals Gonorrhea Gram-Negative Bacteria Growth Hemoglobin Human Immune system Incidence Infection Inflammatory Response Interdisciplinary Study Investigation Iron Iron-Binding Proteins Lactoferrin Leukocyte L1 Antigen Complex Ligand Binding Ligands Lipoproteins Measures Membrane Metabolic Pathway Metabolism Metals Modeling Mucous Membrane Neisseria gonorrhoeae Neutrophil Infiltration Nutritional Nutritional Immunity Nutritional Requirements Parents Pathogenesis Pharyngeal structure Phenotype Postdoctoral Fellow Primates Principal Investigator Proteins Proton-Motive Force Public Health Rectum Research Resistance Role Serum Sexually Transmitted Diseases Siderophores Site Source Superbug Surface System Testing Time Training Transferrin World Health Organization Zinc antimicrobial bacterial genetics career chemokine combat conjunctiva drug-resistant gonorrhea gene product genetic manipulation genome-wide insight interest lactoferrin receptors member microbiota mutant neutrophil novel strategies novel therapeutics pathogen prototype psoriasin receptor reconstruction rectal response skills tenure track transcriptome transcriptome sequencing urogenital tract

项目摘要

PROJECT SUMMARY/ABSTRACT Neisseria gonorrhoeae (Ngo) is the bacterium that causes the sexually transmitted infection gonorrhea. Infection with Ngo triggers a potent inflammatory response at mucosal epithelial surfaces that is characterized by recruitment of neutrophils, which are unable to clear the bacteria. With mounting antibiotic resistance in Ngo, new therapeutics are urgently needed to treat gonorrhea, and one promising approach is to combat its dependence on metals such as iron and zinc that are acquired from its obligate human host. In humans and other mammalian hosts, essential metals are found within high affinity complexes with proteins and are not freely available. Examples of these proteins are the iron-binding transferrin and hemoglobin, found in abundance in serum/erythrocytes, and zinc-binding lactoferrin and calprotectin, which are found in abundance in neutrophils as well as mucosal secretions. To overcome this restriction (“nutritional immunity”), Ngo produces outer-membrane, surface-exposed transporters that bind to human metal-binding proteins and extract the metal from them directly, via energy supplied by the TonB system. There are 8 known TonB- dependent transporters (Tdts) in Ngo. Four Tdts are the focus of the parent application: TdfH and TdfJ, which bind to human calprotectin and psoriasin, respectively, to acquire zinc; and TdfF and TdfG, which are iron- regulated but whose ligands and functions remain undefined. Four other Tdts are involved in metal acquisition in Ngo: TbpA, which binds human transferrin; LbpA, which binds human lactoferrin; HpuB, which binds human hemoglobin; and FetA, which binds ferric enterobactin, a siderophore produced by members of the microbiota. In this diversity supplement application, we propose three Specific Aims that are independent of the parent R01 award: 1) Test the role for TbpAB, FetAB, and HpuAB in survival and outgrowth of Ngo exposed to primary human neutrophils, including the necessity for ligand binding and TonB-dependent transport; 2) Determine the bioavailable metals and their cognate metal-binding proteins that are available to Ngo during infection of neutrophils; and 3) Define the metabolic pathways Ngo differentially uses in metal-depleted and metal-replete growth conditions. The latter Aim will leverage a recent genome-scale metabolic network reconstruction our lab recently created, contextualized with publicly available metal-dependent transcriptional datasets of Ngo. Completion of these Aims will reveal new insights into the response of Ngo to human nutritional immunity and uncover new biology for future mechanistic investigation at this host-pathogen interface. It will provide the diversity supplement postdoctoral trainee with defined training in bacterial genetic manipulation and neutrophil functional assays, while engaging the trainee in the interdisciplinary research taking place under the auspices of the parent R01. At the same time, the trainee will develop career transitional skills and uncover new research foci, which will help her achieve her career goal of becoming an independent tenure-track principal investigator in Ngo biology and pathogenesis.

项目概要/摘要淋病奈瑟菌 (Ngo) 是引起性传播感染淋病的细菌。 Ngo 感染会在粘膜上皮表面引发强烈的炎症反应，其特点是通过募集中性粒细胞，中性粒细胞无法清除细菌，并且抗生素耐药性不断增加。非政府组织，迫切需要新的疗法来治疗淋病，一种有希望的方法是对抗它在人类和人类中，对从其专性人类宿主获得的铁和锌等金属的依赖。与其他哺乳动物宿主相比，必需金属存在于与蛋白质的高亲和力复合物中，而不是这些蛋白质的例子是铁结合转铁蛋白和血红蛋白，存在于中。血清/红细胞中含量丰富，锌结合乳铁蛋白和钙卫蛋白含量丰富为了克服这种限制（“营养免疫”），Ngo 发现了中性粒细胞和粘膜分泌物。产生外膜、表面暴露的转运蛋白，与人类金属结合蛋白结合，通过 TonB 系统提供的能量直接从其中提取金属有 8 种已知的 TonB-。 Ngo 中的依赖转运蛋白 (Tdts) 四个 Tdts 是父申请的重点：TdfH 和 TdfJ，其中分别与人钙卫蛋白和银屑病素结合，获得锌和 TdfF 和 TdfG，它们是铁- 受监管，但其配体和功能仍未定义。另外四个 Tdt 参与金属获取。 Ngo中：TbpA，结合人转铁蛋白；LbpA，结合人乳铁蛋白；HpuB，结合人；血红蛋白；和 FetA，它与铁肠杆菌素（微生物群成员产生的铁载体）结合。在这个多样性补充申请中，我们提出了三个独立于母公司的具体目标 R01 奖：1) 测试 TbpAB、FetAB 和 HpuAB 在暴露于 Ngo 的存活和生长中的作用原代人中性粒细胞，包括配体结合和 TonB 依赖性运输的必要性；2) 确定 Ngo 可以利用的生物可利用金属及其同源金属结合蛋白中性粒细胞感染；3) 定义 Ngo 在金属耗尽和金属耗尽时不同使用的代谢途径后一个目标将利用最新的基因组规模代谢网络。我们的实验室最近创建的重建，与公开的金属依赖性转录相关 Ngo 的数据集的完成将揭示 Ngo 对人类的反应的新见解。营养免疫并揭示新的生物学，用于未来对该宿主病原体的机制研究它将为多样性补充博士后实习生提供细菌遗传方面的明确培训。操作和中性粒细胞功能测定，同时让学员参与跨学科研究在母公司 R01 的主持下进行，同时，实习生将发展职业过渡。技能并发现新的研究焦点，这将帮助她实现成为独立的职业目标非政府组织生物学和发病机制的终身教授首席研究员。

项目成果

期刊论文数量（13）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Stealthy microbes: How Neisseria gonorrhoeae hijacks bulwarked iron during infection.

DOI：
10.3389/fcimb.2022.1017348
发表时间：
2022
期刊：
Frontiers in cellular and infection microbiology
影响因子：
5.7
作者：
通讯作者：

Mutagenesis of the Loop 3 α-Helix of Neisseria gonorrhoeae TdfJ Inhibits S100A7 Binding and Utilization.

DOI：
10.1128/mbio.01670-22
发表时间：
2022-08-30
期刊：
mBio
影响因子：
6.4
作者：
通讯作者：

Transcriptional activation of ompA in Neisseria gonorrhoeae mediated by the XRE family member protein NceR.

DOI：
10.1128/mbio.01244-23
发表时间：
2023-08-31
期刊：
MBIO
影响因子：
6.4
作者：
Holley, Concerta L. L.;Dhulipala, Vijaya;Maurakis, Stavaros A. A.;Greenawalt, Ashley Nicole;Read, Timothy D. D.;Cornelissen, Cynthia N. N.;Shafer, William M. M.
通讯作者：
Shafer, William M. M.

Point Mutations in TbpA Abrogate Human Transferrin Binding in Neisseria gonorrhoeae