The role of surface-bound ectonucleotidase CD73 in modulation of Porphyromonas gingivalis infection in gingival epithelial cells

表面结合核酸外切酶 CD73 在调节牙龈上皮细胞牙龈卟啉单胞菌感染中的作用

• 批准号: 10429903
• 负责人: Jaden Sophien Lee
• 金额: $ 5.18万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10429903
• 关键词: ATP Hydrolysis; Address; Adenosine; Adenosine Monophosphate; Adult; Affect; American; Anti-Inflammatory Agents; Bacterial Infections; Biological Models; Cells; Coupled; Cyclic AMP; Data; Development; Disease; Enzymes; Epithelial; Epithelial Cells; Equilibrium; Event; Feedback; Fellowship; Fostering; Future; Gene Expression; Generations; Germ Cells; Gingiva; Growth; Homeostasis; Host Defense Mechanism; Human; Hydrolysis; Immune; Immune response; Immune signaling; Immunologics; Immunology; Infection; Infection Control; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Interleukin-6; Invaded; Joints; Knowledge; Laboratories; Life; Malignant Neoplasms; Mediating; Mediator of activation protein; Metabolism; Microbe; Molecular; Mucous Membrane; NADPH Oxidase; Oral; Oral Microbiology; Oral cavity; Oral health; Oral mucous membrane structure; Outcome; Pathologic; Pathway interactions; Periodicity; Periodontal Diseases; Physiological; Population; Porphyromonas gingivalis; Production; Reactive Oxygen Species; Regulation; Research; Role; Scientist; Signal Pathway; Signal Transduction; Signaling Molecule; Stat3 protein; Stimulus; Surface; Testing; Time; Tissues; Training; Work; antimicrobial; arm; chemokine; cytokine; extracellular; microbial; microorganism; novel; pathogen; receptor; response; therapeutic development; therapeutically effective

PROJEST SUMMARY/ABSTRACT Danger signal molecule extracellular ATP (eATP) is released by infected cells and becomes metabolized to adenosine by multiple enzymatic steps, one of which is catalyzed by CD73, a surface-bound enzyme that converts adenosine monophosphate (AMP) to adenosine. This AMP hydrolysis by CD73 is an irreversible rate- limiting step in adenosine signaling that is often referred to as “an immunological switch” from pro-inflammatory (eATP) to anti-inflammatory (adenosine) mediator. Porphyromonas gingivalis is a keystone pathogen strongly associated with severe periodontal disease and can use multiple mechanisms to successfully invade, replicate, and disseminate within and through the human gingival epithelial cells (GECs), a major arm of initial defenses in mucosal surfaces. Despite the growing significance in various pathological states including cancer, inflammatory diseases, and recently cellular infection, the role of CD73 for regulating host immune response and bacterial infection in the oral cavity is not well understood. Our laboratory previously showed that P. gingivalis can inhibit eATP-mediated reactive oxygen species (ROS) generation by NADPH oxidase 2 and that the bacterial growth is modulated and enhanced by adenosine 2a receptor-coupled adenosine signaling in GECs. Our novel preliminary findings also revealed significantly increased CD73 expression with P. gingivalis in GECs. Furthermore, CD73-mediated signaling specifically impacted on the P. gingivalis intracellular survival, inhibited ROS generation, and dampened the gene expression of interleukin-6 (IL-6) whose addition led to decreased levels of P. gingivalis in GECs. Thus, our overarching hypothesis is that P. gingivalis can selectively regulate host CD73-mediated immune signaling pathways for affluent intracellular growth and survival in the gingival epithelium. We will test our hypothesis through completion of the following specific aims. Specific Aim 1 will establish the regulatory role of CD73 for P. gingivalis intracellular infection by further investigating functional importance supporting the P. gingivalis/CD73 coupled signaling in the GECs. Specific Aim 2 will elucidate select cellular and molecular mechanisms by which CD73 modulates P. gingivalis intracellular growth and survival, specifically through interfering with pro-inflammatory IL-6 signaling. Both aims will use primary GECs model system to functionally dissect out mechanisms and define the molecular events. Hence, this proposal will characterize novel physiologically relevant modulator(s) of oral bacterial infection and reveal the multidirectional cross-talk between host CD73/P. gingivalis/IL-6 axis for the modulation of intracellular P. gingivalis infection in the host cells. The results of this study will provide fundamentally novel molecular understanding of P. gingivalis persistence mechanisms in the oral mucosa and may ultimately contribute to future development of effective therapeutics for controlling P. gingivalis associated infections. This fellowship will also provide training in oral microbiology/immunology and will foster the development of the trainee into a unique and critically needed oral health academic clinician/scientist.

Projest摘要/摘要 危险信号分子细胞外ATP（EATP）被感染的细胞释放，并被代谢为 腺苷通过多个酶促步骤，其中之一是由CD73催化的，CD73是一种表面结合的酶 将腺苷一磷酸（AMP）转换为腺苷。 CD73的这种AMP水解是一种不可逆的速率 - 限制腺苷信号的步骤，通常称为促炎的“免疫转换” （EATP）到抗炎（腺苷）介体。牙龈卟啉单胞菌是一种强烈的基石病原体 与严重的牙周疾病有关，可以使用多种机制成功入侵，重复， 并通过人牙龈上皮细胞（GEC）传播，这是初始防御的主要部门 在粘膜表面。尽管在包括癌症在内的各种病理状态中具有越来越重要的意义，但 炎症性疾病和最近的细胞感染，CD73在控制宿主免疫反应和 口腔中的细菌感染尚不清楚。我们的实验室先前表明牙龈疟原虫 可以抑制NADPH氧化蛋白酶2的EATP介导的活性氧（ROS）2，并且细菌 GEC中的腺苷2A受体偶联腺苷信号传导调节生长并增强生长。我们的小说 初步发现还显示，在GEC中，CD73表达显着增加。 此外，CD73介导的信号转导对牙龈疟原虫细胞内存活有特异性影响，抑制了 ROS产生，并诅咒白介素6（IL-6）的基因表达，其增加导致下降 GEC中牙龈疟原虫的水平。这就是我们的总体假设是牙龈疟原虫可以选择性地 调节宿主CD73介导的免疫信号通路，以影响细胞内生长和存活率 在牙龈上皮中。我们将通过完成以下特定目标来检验我们的假设。具体的 AIM 1将通过进一步研究CD73在细胞内感染中的调节作用 功能重要性支持GEC中的牙龈疟原虫/CD73耦合信号传导。具体目标2将 阐明CD73调节细胞内生长的精选细胞和分子机制 和生存，特别是通过干扰促炎性IL-6信号传导。两个目标都将使用主要 GECS模型系统可以在功能上剖析机制并定义分子事件。因此，这个 建议将表征口腔细菌感染的新型物理相关调节剂 揭示宿主CD73/p之间的多向串扰。牙龈/IL-6轴以调制 宿主细胞中细胞内牙龈疟原虫感染。这项研究的结果将提供根本新颖的 对口服粘膜中牙龈疟原虫持续机制的分子理解，最终可能 有助于未来开发有效的疗法来控制牙龈疟原虫相关的感染。这 奖学金还将提供口腔微生物学/免疫学培训，并将促进受训者的发展 成为一个独特而急需的口腔健康学术临床/科学家。