P. gingivalis: Role of GSK3 in Host Inflammation

牙龈卟啉单胞菌：GSK3 在宿主炎症中的作用

• 批准号: 8055392
• 负责人: David A Scott
• 金额: $ 28.11万
• 依托单位: UNIVERSITY OF LOUISVILLE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-25 至 2012-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8055392
• 关键词: Abscess; Affect; Alveolar Bone Loss; Anti-Inflammatory Agents; Anti-inflammatory; Attenuated; Cell Nucleus; Cell surface; Cells; Chronic; Dendritic Cells; Disease; Event; Foundations; Gene Expression; Glycogen Synthase Kinase 3; Host Defense; Immune; Immune response; Immune system; Immunity; Inflammation; Inflammatory; Inflammatory Response; Interferons; Interleukin-1; Interleukin-10; Interleukin-12; Interleukin-6; Invaded; Laboratories; Mediating; Microbe; Modeling; Mus; Nature; Organism; Pathogenesis; Pathway interactions; Periodontal Diseases; Phosphotransferases; Play; Porphyromonas gingivalis; Principal Investigator; Process; Production; Property; Protein Isoforms; Receptor Activation; Receptor Signaling; Regulation; Reporting; Role; Severity of illness; Signal Pathway; Signal Transduction; Signaling Molecule; Solid; TLR2 gene; TLR4 gene; Toll-like receptors; base; cytokine; design; in vivo; inhibitor/antagonist; macrophage; microorganism interaction; mouse model; pathogen; programs; response; therapeutic target

DESCRIPTION (provided by applicant): The host immune response to periodontal-associated pathogens has been shown to be a key determinant involved in periodontal disease by regulating the production of pro- and anti-inflammatory cytokines that regulate the severity of this disease. An inability to regulate the nature or duration of the host's inflammatory response can often mediate detrimental host effects as observed in chronic inflammatory diseases. Our laboratory has identified the kinase glycogen synthase kinase 3 (GSK3) as a central regulator of the inflammatory process by its ability to differentially regulate the levels of pro- and anti-inflammatory cytokines upon TLR activation. Our specific hypothesis is that the ability of P. gingivalis to interact with TLR2 and TLR4 expressed on immune cells and the subsequent differential regulation of GSK3 activity mediated by these TLRs are major mechanisms responsible for the pathogenesis of this organism by regulating the nature and magnitude of the host's inflammatory response. This hypothesis is based on: 1) stimulation of immune cells from TLR2- or TLR4-deficient mice with P. gingivalis resulted in pronounced differences in the inhibition of GSK3; 2) inhibition of GSK3 in macrophages or dendritic cells stimulated with P. gingivalis resulted in the severe reduction in the levels of pro-inflammatory cytokines, i.e. IL-12, while concurrently augmenting the levels of the anti-inflammatory cytokine IL-10; 3) a direct comparison of TLR2- and TLR4- deficient cells stimulated with P. gingivalis demonstrated that the levels of pro-inflammatory cytokines such as IL-12 were largely mediated by TLR4 whereas the levels of the anti-inflammatory cytokine IL-10 were predominantly mediated by TLR2; 4) analysis of the two major isoforms of GSK3 in innate immune cells revealed that dendritic cells contain only detectable levels of GSKS-p, whereas macrophages contain both isoforms of GSK3, GSK3-a and GSK3-p; 5) in vivo administration of a GSK3 inhibitor resulted in a potent reduction in the levels of IL-1b, IL-6, IL-12, and IFN-y whereas the levels of the anti-inflammatory cytokine IL-10 were potently augmented in response to P. gingivalis; and 6) inhibition of GSK3 in mice resulted in a predominant Th2-assoicated immune response to P. gingivalis. These observations provide a strong rationale and focus concerning the reported differences of TLR2 and TLR4 to mediate pro- and anti- inflammatory cytokine responses and elucidate a critical role for the GSK3 cell-signaling pathway in modulating the host's inflammatory response to P. gingivalis. The specific aims are designed to characterize how host-microbial interactions are involved in the regulation of the host inflammatory response to P. gingivalis. to elucidate and characterize the involvement of GSK3 in regulating these immune responses, and to evaluate the efficacy of a therapeutic target (GSK3) that could attenuate the disease process in vivo.

描述（由申请人提供）：通过调节调节这种疾病严重程度的促促血细胞因子的产生，宿主对牙周相关病原体的免疫反应已被证明是牙周疾病中涉及的关键决定因素。无法调节宿主炎症反应的性质或持续时间通常会介导慢性炎症性疾病中观察到的有害宿主作用。我们的实验室通过在TLR激活后差异调节促促促促促血管和抗炎细胞因子的水平，将激酶糖原合酶3（GSK3）确定为炎症过程的中心调节剂。我们的具体假设是，在免疫细胞上表达的TLR2和TLR4与TLR2和TLR4相互作用的能力以及随后由这些TLR介导的GSK3活性的差异调节是负责这种生物体发病机理的主要机制，通过调节宿主炎症反应的性质和大小。该假设基于：1）刺激具有牙龈疟原虫的TLR2-或TLR4缺陷小鼠的免疫细胞，导致抑制GSK3的明显差异； 2）抑制巨噬细胞或牙龈疟原虫刺激的树突状细胞中GSK3的抑制作用导致促炎性细胞因子的水平严重降低，即IL-12，同时同时增加抗炎细胞因子IL-10的水平； 3）用牙龈疟原虫刺激的TLR2-和TLR4缺陷细胞的直接比较表明，促炎性细胞因子（例如IL-12）的水平在很大程度上是由TLR4介导的，而抗炎性细胞因子IL-10的水平主要由TLR2介导。 4）对先天免疫细胞中GSK3的两种主要同工型的分析表明，树突状细胞仅包含可检测水平的GSKS-P，而巨噬细胞均包含GSK3，GSK3-A和GSK3-P的同工型。 5）在体内给药GSK3抑制剂导致IL-1B，IL-6，IL-12和IFN-Y的水平有效降低，而抗炎细胞因子IL-10的水平则迅速增强，以响应于P. gingivalis； 6）在小鼠中抑制GSK3导致对牙龈疟原虫的占主导地位的免疫反应。这些观察结果提供了强烈的基本原理和重点，涉及TLR2和TLR4的差异，以介导促炎和抗炎细胞因子反应，并阐明了GSK3细胞信号途径在调节宿主对gingivalis炎症性host的炎症反应中的关键作用。该特定目的旨在表征宿主 - 微生物相互作用如何与宿主对牙龈疟原虫的炎症反应有关。为了阐明和表征GSK3在调节这些免疫反应中的参与，并评估可以在体内减弱疾病过程的治疗靶标（GSK3）的疗效。