A. Actinomycetemcomitans Cdt Induces Pro-Inflammatory Innate Immune Responses

A.放线菌 Cdt 诱导促炎症先天免疫反应

• 批准号: 10640870
• 负责人: Kathleen Boesze-Battaglia
• 金额: $ 52.27万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-04-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10640870
• 关键词: Actinobacillus actinomycetemcomitans; Antigens; Apoptosis; Apoptotic; Arachidonic Acids; Binding; Carrier Proteins; Caspase; Cell Cycle Arrest; Cell Line; Cell Proliferation; Cell physiology; Cells; Cessation of life; Cholesterol; Chronic; Complex; Defect; Dinoprostone; Disease; Eicosanoids; Endosomes; Epithelial Cells; Event; Exhibits; Goals; Heat shock proteins; Heat-Shock Proteins 90; Host Defense; Human; IL18 gene; Immunotoxins; Impairment; Individual; Infection; Inflammasome; Inflammation; Inflammation Mediators; Inflammatory; Inflammatory Response; Inflammatory Response Pathway; Innate Immune Response; Interleukin-6; Intoxication; Investigation; Knowledge; Link; Lipids; Lymphocyte; Lytic; Macrophage; Mediating; Membrane Lipids; Membrane Microdomains; Microbe; Molecular; Movement; Organism; Outcome; PI3 gene; PIK3CG gene; Pathogenesis; Pathogenicity; Pathway interactions; Periodontal Diseases; Periodontitis; Phagocytes; Phagocytosis; Phagolysosome; Phagosomes; Phosphatidylinositols; Phosphoric Monoester Hydrolases; Phosphorylation; Play; Production; Property; Prostaglandin-Endoperoxide Synthase; Proteins; Research; Role; Signal Pathway; Signal Transduction; Sorting; System; Targeted Toxins; Testing; Therapeutic Intervention; Thromboxanes; Toxic effect; Toxin; Translating; Virulence Factors; adaptive immune response; cell type; cellugyrin; cytokine; cytolethal distending toxin; holotoxins; insight; microbial; monocyte; novel; oral pathogen; pathogen; repaired; response; retrograde transport; small molecule; spatiotemporal; synaptogyrin; trafficking; tripolyphosphate; vesicle transport; virtual

Periodontal disease is a chronic inflammatory disorder driven by polymicrobial infection. The pathogenesis of this disorder involves activation, and possibly perturbation, of both innate and adaptive immune responses. Over the course of our investigation we have demonstrated that Aggregatibacter actinomycetemcomitans (Aa), a putative pathogen implicated in the pathogenesis of localized aggressive periodontitis (LAP), produces an immunotoxin, the cytolethal distending toxin (Cdt). Cdt is a heterotrimeric holotoxin which functions as an AB2 toxin: the cell binding (B) units are comprised of the CdtA and CdtC subunits and the active (A) subunit, CdtB. We have demonstrated that CdtB functions as a 5'-phosphatidylinositol (PI) triphosphate phosphatase as it degrades the PI-3K signaling lipid, PI-3,4,5-triphosphate (PIP3) to PI-3,4P2; this action leads to blockade of the signaling pathway. Blockade of PI-3K signaling in human macrophages leads to a pro-inflammatory cytokine response involving both canonical and noncanonical inflammasome activation. Cdt also induces production of inflammatory mediators derived from arachidonic acid including PGE2 and thromboxanes. Most recently, we have observed phagocytic defects in Cdt-treated macrophages consistent with altered phago-lysosome maturation. We propose that Cdt perturbs macrophage function thereby contributing to both inflammation and sustained infection. Our overarching hypothesis is that the Aa Cdt contributes to altered local host defense which facilitate Aa survival and enables other microbes to evade host defense. The goal of our study is to extend our investigation and advance our knowledge of the molecular events that link PI-3K signaling blockade to downstream pro- inflammatory responses and altered vesicular trafficking and fusion. Specifically, we propose that CdtB induces GSK3â-dependent HSP90 activation and further that HSP90, via its effects on NLRP3, Cox-2 and possibly ESCRT proteins is a critical intermediary in events leading to release of mature cytokines and eicosanoids (specific aim 1). We plan to advance our understanding of the role that Cdt plays activating the noncanonical inflammasome which involve gasdermin D (GSDMD) pore formation and pyroptosis. Our focus will be to determine how CdtB activates caspase-4 and contributes to GSDMD pore formation; the latter studies will focus on PI-3,4P2 and pore repair via the ESCRT system (specific aim2). In specific aim 3 we will focus on perturbation of vesicular transport and phago-lysosome formation. We propose that altered PI distribution due in part to CdtB-mediated production of PI3,4P2 facilitates CdtB retrograde transport and modulates the formation of phago-lysosomes. The long-term goals of our study are to translate our understanding of the molecular events that govern Cdt toxicity and, in turn, the pathogenicity of Cdt-producing organisms. These studies are of particular significance as Cdt is produced not only by Aa but many other pathogens that contribute to chronic infectious and inflammatory disorders. Insight gained through this investigation will advance our understanding of the molecular events underlying LAP and other diseases caused by Cdt-producing organisms and identify new avenues for therapeutic intervention. .

牙周病是一种由多种微生物感染引起的慢性炎症性疾病。发病机制 这种疾病的发生涉及先天性和适应性免疫反应的激活，甚至可能的扰动。超过 在我们的调查过程中，我们已经证明，Aggregatibacter actinomycetemcomitans (Aa) 是一种 假定的病原体与局部侵袭性牙周炎（LAP）的发病机制有关，产生一种 免疫毒素，细胞致死膨胀毒素（Cdt）。 Cdt 是一种异三聚体全毒素，其功能类似于 AB2 毒素：细胞结合 (B) 单位由 CdtA 和 CdtC 亚单位以及活性 (A) 亚单位 CdtB 组成。 我们已经证明 CdtB 作为 5'-磷脂酰肌醇 (PI) 三磷酸磷酸酶发挥作用，因为它 将 PI-3K 信号脂质 PI-3,4,5-三磷酸 (PIP3) 降解为 PI-3,4P2；这一行动导致封锁 信号通路。阻断人类巨噬细胞中的 PI-3K 信号传导会产生促炎细胞因子 涉及典型和非典型炎症小体激活的反应。 Cdt 还诱导产生 源自花生四烯酸的炎症介质，包括 PGE2 和血栓素。最近，我们有 在 Cdt 处理的巨噬细胞中观察到吞噬细胞缺陷，这与吞噬溶酶体成熟的改变一致。我们 提出 Cdt 扰乱巨噬细胞功能，从而导致炎症和持续感染。 我们的首要假设是 Aa Cdt 有助于改变本地宿主防御，从而促进 Aa 生存并使其他微生物能够逃避宿主防御。我们研究的目的是扩大我们的调查范围 并增进我们对将 PI-3K 信号传导阻断与下游亲连接起来的分子事件的了解 炎症反应和改变的囊泡运输和融合。具体来说，我们建议 CdtB 诱导 GSK3â 依赖性 HSP90 激活以及 HSP90 通过其对 NLRP3、Cox-2 和可能的 ESCRT 的影响 蛋白质是导致成熟细胞因子和类二十烷酸释放的事件中的关键中介（具体目标 1）。我们计划加深对 Cdt 激活非典型炎症小体作用的理解 其中涉及gasdermin D (GSDMD) 孔形成和细胞焦亡。我们的重点是确定 CdtB 如何 激活 caspase-4 并有助于 GSDMD 孔形成；后期研究将重点关注PI-3,4P2和孔隙 通过 ESRT 系统进行修复（特定目标2）。在具体目标 3 中，我们将重点关注囊泡运输的扰动 和吞噬溶酶体的形成。我们认为 PI 分布的改变部分是由于 CdtB 介导的产生 PI3,4P2 促进 CdtB 逆行转运并调节吞噬溶酶体的形成。长期来看 我们研究的目标是转化我们对控制 Cdt 毒性的分子事件的理解，进而 产生 Cdt 的微生物的致病性。这些研究具有特别重要的意义，因为 Cdt 的产生不是 仅由 Aa 引起，但许多其他病原体也会导致慢性感染和炎症性疾病。洞察力 通过这项研究获得的成果将加深我们对 LAP 和其他分子事件的理解 产生 Cdt 的生物体引起的疾病并确定治疗干预的新途径。 。

项目成果

Aggregatibacter actinomycetemcomitans Cytolethal Distending Toxin-Induces Cell Cycle Arrest in a Glycogen Synthase Kinase (GSK)-3-Dependent Manner in Oral Keratinocytes.

• DOI: 10.3390/ijms231911831
• 发表时间: 2022-10-05
• 期刊: International journal of molecular sciences
• 影响因子: 5.6

The Active Subunit of the Cytolethal Distending Toxin, CdtB, Derived From Both Haemophilus ducreyi and Campylobacter jejuni Exhibits Potent Phosphatidylinositol-3,4,5-Triphosphate Phosphatase Activity.

• DOI: 10.3389/fcimb.2021.664221
• 发表时间: 2021
• 期刊: Frontiers in cellular and infection microbiology
• 影响因子: 5.7
• 作者: Huang G;Boesze-Battaglia K;Walker LP;Zekavat A;Schaefer ZP;Blanke SR;Shenker BJ
• 通讯作者: Shenker BJ

The Cell-Cycle Regulatory Protein p21CIP1/WAF1 Is Required for Cytolethal Distending Toxin (Cdt)-Induced Apoptosis.

细胞周期调节蛋白 p21CIP1/WAF1 是细胞致死膨胀毒素 (Cdt) 诱导的细胞凋亡所必需的。

• DOI: 10.3390/pathogens9010038
• 发表时间: 2020
• 期刊: Pathogens (Basel, Switzerland)
• 作者: Shenker,BruceJ;Walker,LisaM;Zekavat,Ali;Weiss,RobertH;Boesze-Battaglia,Kathleen
• 通讯作者: Boesze-Battaglia,Kathleen