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

基本信息

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
项目类别：
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

项目摘要

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. .

牙周疾病是由多数菌感染驱动的慢性炎症性疾病。发病机理这种疾病涉及先天和适应性免疫调查的激活和可能的扰动。超过我们的调查过程表明，聚集的放线菌菌（AA），a 在局部攻击性牙周炎（LAP）发病机理中实施的假定病原体，产生免疫毒素，细胞杀伤的毒素（CDT）。 CDT是一种异三聚毒素，可作为AB2起作用毒素：细胞结合（b）单位是CDTA和CDTC亚基的积累，以及活性（a）亚基CDTB。我们已经证明CDTB充当5'-磷脂酰肌醇（PI）三磷酸磷酸酶将PI-3K信号脂质降解为PI-3,4,5-三磷酸盐（PIP3）至PI-3,4P2；此行动导致了封锁信号通路。人类巨噬细胞中PI-3K信号传导的封锁导致促炎性细胞因子反应涉及规范和非规范炎症体激活。 CDT还引起了生产源自包括PGE2和血栓烷的花生四烯酸的炎症介质。最近，我们有观察到的吞噬细胞缺陷在经CDT处理的巨噬细胞中，与phago-lysosomys体成熟的改变一致。我们提议CDT伴有巨噬细胞功能，从而导致炎症和持续感染。我们的总体假设是AA CDT有助于改变当地的主机防御 AA生存，使其他微生物能够逃避宿主防御。我们研究的目的是扩展我们的调查并促进我们对将PI-3K信号阻滞与下游促进联系起来的分子事件的了解炎症反应和囊泡运输和融合改变。具体而言，我们建议CDTB诱导 GSK3依赖性HSP90激活，进一步的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花生酸的事件中的关键中间体（特定目的 1）。我们计划促进我们对CDT扮演非规范炎症体的作用的理解涉及Gasdermin D（GSDMD）孔形成和凋亡。我们的重点是确定CDTB 激活caspase-4并有助于GSDMD孔的形成；后者的研究将重点放在PI-3,4P2和毛孔上通过ESCRT系统维修（特定AIM2）。在特定的目标3中，我们将专注于囊泡运输的扰动和Phago莱斯体形成。我们建议改变PI分布部分是由于CDTB介导的生产 PI3,4P2的最爱CDTB逆行转运，并调节了Phago-lysosomes的形成。长期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，然后又产生CDT的生物的致病性。这些研究特别重要，因为没有产生CDT 仅通过AA，但许多其他病原体会导致慢性感染和炎症性疾病。洞察力通过这项调查获得的将提高我们对圈和其他分子事件的理解由产生CDT的生物引起的疾病，并确定用于治疗干预的新途径。。

项目成果

期刊论文数量（7）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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
作者：
通讯作者：

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)
影响因子：
0
作者：
Shenker,BruceJ;Walker,LisaM;Zekavat,Ali;Weiss,RobertH;Boesze-Battaglia,Kathleen
通讯作者：
Boesze-Battaglia,Kathleen

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