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）：宿主对牙周相关病原体的免疫反应已被证明是参与牙周病的关键决定因素，通过调节促炎和抗炎细胞因子的产生来调节该疾病的严重程度。无法调节宿主炎症反应的性质或持续时间通常会介导有害的宿主效应，如在慢性炎症性疾病中观察到的那样。我们的实验室已将激酶糖原合酶激酶 3 (GSK3) 确定为炎症过程的中心调节因子，因为它能够在 TLR 激活时差异调节促炎细胞因子和抗炎细胞因子的水平。我们的具体假设是，牙龈卟啉单胞菌与免疫细胞上表达的 TLR2 和 TLR4 相互作用的能力以及随后由这些 TLR 介导的 GSK3 活性的差异调节是该生物体发病机制的主要机制，通过调节免疫细胞上表达的 TLR2 和 TLR4 的性质和程度。宿主的炎症反应。该假设基于：1）用牙龈卟啉单胞菌刺激 TLR2 或 TLR4 缺陷小鼠的免疫细胞，导致 GSK3 的抑制作用显着差异； 2)用牙龈卟啉单胞菌刺激的巨噬细胞或树突状细胞中GSK3的抑制导致促炎细胞因子（即IL-12）的水平严重降低，同时增加抗炎细胞因子IL-10的水平； 3) 直接比较用牙龈卟啉单胞菌刺激的 TLR2 和 TLR4 缺陷细胞表明，促炎细胞因子（如 IL-12）的水平主要由 TLR4 介导，而抗炎细胞因子 IL-10 的水平则主要由 TLR4 介导。主要由 TLR2 介导； 4) 对先天免疫细胞中 GSK3 的两种主要亚型的分析表明，树突状细胞仅含有可检测水平的 GSKS-p，而巨噬细胞则含有 GSK3、GSK3-a 和 GSK3-p 两种亚型； 5) 体内施用 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 的水平则有效增强。对牙龈卟啉单胞菌的反应； 6) 抑制小鼠中的 GSK3 导致针对牙龈卟啉单胞菌的主要 Th2 相关免疫反应。这些观察结果为所报道的 TLR2 和 TLR4 在介导促炎和抗炎细胞因子反应方面的差异提供了强有力的理由和焦点，并阐明了 GSK3 细胞信号传导途径在调节宿主对牙龈卟啉单胞菌的炎症反应中的关键作用。具体目标旨在描述宿主-微生物相互作用如何参与调节宿主对牙龈卟啉单胞菌的炎症反应。阐明和表征 GSK3 在调节这些免疫反应中的作用，并评估可以减轻体内疾病过程的治疗靶点 (GSK3) 的功效。