Novel carbohydrate binding functions of the CR3 I-domain modulate gonococcal-cervical cell interactions

CR3 I 结构域的新型碳水化合物结合功能调节淋球菌-宫颈细胞相互作用

• 批准号: 10318111
• 负责人: Jennifer L Edwards
• 金额: $ 38.75万
• 依托单位: RESEARCH INST NATIONWIDE CHILDREN'S HOSP
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-16 至 2023-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10318111
• 关键词: Adhesions; Affect; Agonist; Bacteria; Bacterial Proteins; Behavior; Binding; Binding Sites; Biological; C3bi; Calorimetry; Carbohydrates; Cell Communication; Cells; Centers for Disease Control and Prevention (U.S.); Cervical; Cervicitis; Cervix Uteri; Cessation of life; Coculture Techniques; Complement; Data; Development; Diagnosis; Disease; Ensure; Epithelial; Epithelial Cells; Event; Female; Fimbriae Proteins; Foundations; Frequencies; Future; Goals; Gonorrhea; HIV Infections; Health; Human; Immune; Immune response; Immune system; Immunity; Immunologic Receptors; In Vitro; Infection; Infertility; Innate Immune Response; Lectin; Ligands; Link; Literature; Macrophage-1 Antigen; Mass Spectrum Analysis; Mediating; Mediator of activation protein; Medical Care Costs; Molecular; Morbidity - disease rate; Mucosal Immunity; Mucous Membrane; Multi-Drug Resistance; Natural Immunity; Nature; Neisseria gonorrhoeae; Outcome; Pathogenesis; Pathway interactions; Pattern recognition receptor; Pelvic Inflammatory Disease; Peptides; Phagocytes; Phase; Pilum; Play; Polysaccharides; Post-Translational Protein Processing; Prevalence; Prevention; Preventive measure; Process; Proteins; Public Health; Recombinants; Reporting; Role; Seminal; Signal Transduction; Structure; Surface; Surface Plasmon Resonance; Testing; Therapeutic; United States; Ursidae Family; Variant; Woman; Work; appendage; base; experience; glycosylation; human disease; human pathogen; improved; innate immune function; innovation; insight; macrophage; microbial; neutrophil; novel; novel strategies; novel therapeutics; pathogen; pathogenic bacteria; prevent; receptor; reproductive tract; response; sugar; trafficking; vaccine development

PROJECT SUMMARY The prevalence of Neisseria gonorrhoeae (Ng), its associated morbidity, and the emergence of untreatable strains, support a critical need for new preventative and therapeutic strategies for this important human- pathogen. Such developments require a complete understanding of the pathobiology of this archetypal, host- adapted pathogen. We show that phase-variable, glycan post-translational modifications on Ng surface appendages, pili (fimbriae), play a vital role in human infection. Complement receptor 3 (CR3) is an important pattern recognition receptor that is the key receptor mediating Ng colonization of human cervical mucosa. CR3 contains an “I-domain” region, which is known as a binding site for protein ligands. However, we found that Ng binding to the I-domain is mediated by the pilin-linked glycan (PLG). This is a seminal finding in innate immunity, as all previous literature ascribe CR3 carbohydrate binding to a separate, so called "lectin" domain. This was also the first demonstration that post-translational modifications made to a bacterial protein modulate pathogenesis. These findings will impact our understanding of microbial pathogenesis and innate immune responses. The goals of the present application are to define the specific contribution of the six, naturally occurring, PLG structures in mediating the Ng-CR3 I-domain interaction and to determine the biological relevance of each of these interactions to infection in females. Guided by strong preliminary data, we hypothesize that variable PLG structures initiate key, but highly divergent, outcomes with CR3 I-domain engagement. We will resolve our hypothesis through two specific aims: Aim 1) Define the effect of variation in pilin glycan structure on direct PLG-I-domain interactions; we will define the specific molecular interactions occurring with the CR3 I-domain for the six natural Ng PLG structures. Aim 2) Define the effect of PLG-I- domain interactions on Ng pathogenesis and CR3 function; we will define targeted epithelial cell responses to CR3 I-domain engagement by Ng that bear different PLG structures and their effect on Ng pathogenesis. Our approach is innovative in using biologically relevant human primary cervical cells, both alone and in co-culture with phagocytes, combined with low passage Ng isolates. This ensures that data obtained are relevant to human processes. By defining the effector functions controlled by I-domain lectin activity on primary human epithelial cells, we will provide critical new information regarding Ng pathogenesis, CR3 function, and the fundamental mechanisms that govern human cervical mucosal immunity. Moreover, we will define the utility of targeting the PLG-CR3 I-domain interaction as a new, and improved, host-targeted approach to treat and/or prevent Ng disease in women. Our studies will very likely impact work on other human pathogens and on innate immunity, broadening the significance of our outcomes. We have extensive experience in defining the molecular mechanisms of Ng pathogenesis. We are ideally placed to do this work in having discovered CR3 as the key receptor for Ng cervical infection and the biosynthetic pathways for pili glycosylation.

项目总结 淋病奈瑟菌(Ng)的流行、相关发病率和无法治疗的出现 菌株，支持对这种重要的人类-新的预防和治疗策略的迫切需要- 病原体。这样的发展需要对这种原型宿主的病理生物学有一个完整的了解。 适应病原体。我们证明了Ng表面上的相变、糖链的平移后修饰 附属物，菌毛(菌毛)，在人类感染中起着至关重要的作用。补体受体3(CR3)是一种重要的 模式识别受体是介导Ng在人宫颈黏膜定植的关键受体。CR3 含有一个“I-结构域”区域，它被称为蛋白质配体的结合部位。但是我们发现， NG与I-结构域的结合是由菌毛素连接的多糖(PLG)介导的。这是《天生》杂志的一个开创性发现 免疫，正如所有以前的文献将CR3碳水化合物结合到一个单独的，所谓的凝集素结构域。 这也是第一次证明了细菌蛋白质的翻译后修饰。 调节病机。这些发现将影响我们对微生物发病机制和先天遗传的理解。 免疫反应。本申请的目的是定义这六个人的具体贡献， 自然发生的PLG结构在介导Ng-CR3 I结构域相互作用中的作用 这些相互作用中的每一个与女性感染的生物学相关性。在强劲的初步数据指引下，我们 假设可变PLG结构启动CR3I-结构域的关键但高度不同的结果 订婚。我们将通过两个具体目标来解决我们的假设：目标1)定义变化的影响 PLG-I结构域直接相互作用上的菌胶多聚糖结构；我们将定义特定的分子相互作用 与六个天然Ng PLG结构的CR3I-结构域一起发生。目的2)定义PLG-I的作用- 结构域相互作用在Ng发病机制和CR3功能中的作用；我们将定义靶向上皮细胞对 具有不同PLG结构的Ng对CR3I结构域的参与及其在Ng发病中的作用。我们的 该方法在使用生物相关的人类原代宫颈细胞方面具有创新性，无论是单独培养还是共同培养 吞噬细胞，与低传代Ng分离株结合。这确保了所获得的数据与 人类的过程。通过定义原始人I-结构域凝集素活性控制的效应器功能 上皮细胞，我们将提供关于Ng发病机制、CR3功能和 控制人类宫颈粘膜免疫的基本机制。此外，我们还将定义效用 将PLG-CR3 I结构域相互作用作为一种新的、改进的宿主靶向治疗和/或 预防女性Ng病。我们的研究很可能会影响对其他人类病原体的研究 与生俱来的免疫力，扩大了我们结果的意义。我们有丰富的经验来定义 Ng发病的分子机制。我们发现CR3是完成这项工作的理想位置 Ng宫颈感染的关键受体和菌毛糖基化的生物合成途径。

项目成果

Complement Receptor 3 Mediates HIV-1 Transcytosis across an Intact Cervical Epithelial Cell Barrier: New Insight into HIV Transmission in Women.

• DOI: 10.1128/mbio.02177-21
• 发表时间: 2022-02-22
• 期刊: mBio
• 影响因子: 6.4
• 作者: Day CJ;Hardison RL;Spillings BL;Poole J;Jurcisek JA;Mak J;Jennings MP;Edwards JL
• 通讯作者: Edwards JL

Complete Genome Sequence of Serogroup B Neisseria meningitidis Strain C311.

• DOI: 10.1128/mra.00788-21
• 发表时间: 2021-10-21
• 期刊: Microbiology resource announcements
• 影响因子: 0.8
• 作者: Jen FE;Atack JM;Zhang Y;Edwards JL;Jennings MP
• 通讯作者: Jennings MP

Analysis of Bacterial Phosphorylcholine-Related Genes Reveals an Association between Type-Specific Biosynthesis Pathways and Biomolecules Targeted for Phosphorylcholine Modification.

• DOI: 10.1128/spectrum.01583-23
• 发表时间: 2023-08-17
• 期刊: MICROBIOLOGY SPECTRUM
• 影响因子: 3.7
• 作者: Zhang, Yuan;Jen, Freda E. -C.;Edwards, Jennifer L. L.;Jennings, Michael P. P.
• 通讯作者: Jennings, Michael P. P.

Complete Genome Sequences of Seven Neisseria gonorrhoeae Clinical Isolates from Mucosal and Disseminated Gonococcal Infections.

• DOI: 10.1128/mra.00734-21
• 发表时间: 2021-10-28
• 期刊: Microbiology resource announcements
• 影响因子: 0.8
• 作者: Jen FE;Atack JM;Edwards JL;Jennings MP
• 通讯作者: Jennings MP