Dominant microRNAs as biomarkers in innate immunity and periodontitis

主要 microRNA 作为先天免疫和牙周炎生物标志物

• 批准号: 10063992
• 负责人: EDWARD K CHAN
• 金额: $ 38.13万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-12-01 至 2024-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10063992
• 关键词: Affect; Alzheimer' s Disease; Amyloidosis; Autoimmune Diseases; Bacteria; Biological Assay; Biological Markers; Biology; Cardiovascular Diseases; Cells; Cellular biology; Chronic; Clinical; Clinical Research; Complex; Data; Dendritic Cells; Dental Cementum; Dental Plaque; Development; Disease; Disease Management; Ecology; Endothelium; Endotoxins; Epithelial; Epithelial Cells; Fibroblasts; Forsythia; Fusobacterium nucleatum; Future; Gene Expression; Generations; Gingiva; Gingival Crevicular Fluid; Gingival Pocket; Gingivitis; Goals; Hemophilia A; Human; Hybrids; IRAK4 gene; Immune signaling; In Vitro; Individual; Infection; Inflammation; Inflammatory; Inherited; Innate Immune Response; Journals; Kinetics; Knockout Mice; LDL Cholesterol Lipoproteins; Laboratories; Ligation; Lipopolysaccharides; Malignant Neoplasms; Maxilla; Mediating; Messenger RNA; MicroRNAs; Modeling; Molecular Biology; Mus; Myelogenous; Natural Immunity; Oral; Osteoblasts; Osteoclasts; Pathogenesis; Pathway interactions; Patients; Peer Review; Peptidoglycan; Periodontal Diseases; Periodontal Ligament; Periodontitis; Play; Porphyromonas gingivalis; Prealbumin; Predisposition; Production; Protocols documentation; Publishing; Regulation; Reporting; Resistance; Rheumatoid Arthritis; Role; Saliva; Series; Severity of illness; Signal Pathway; Signal Transduction; Small RNA; Spleen; Streptococcus gordonii; System; Systemic disease; TLR1 gene; TLR2 gene; TLR4 gene; TNF Receptor-Associated Factors; Therapeutic Intervention; Time; Tissues; Toll-Like Receptor Pathway; Toll-like receptors; Treponema denticola; Virus Diseases; Work; adverse pregnancy outcome; alveolar bone; base; clinically relevant; conventional therapy; crosslink; cytokine; design; dysbiosis; improved; in vivo; individual patient; innate immune pathways; knockout gene; mRNA Expression; macrophage; microbial; monocyte; mouse model; neutrophil; novel therapeutics; oral cavity epithelium; pathogen; pathogenic bacteria; periodontopathogen; recruit; response; therapeutic target; therapy outcome

Abstract Periodontal disease affects millions of individuals in the US alone and has been substantiated as a precursor to other debilitating systemic diseases, including cardiovascular disease, Alzheimer’s disease, rheumatoid arthritis, and adverse pregnancy outcomes. Our laboratories have shown that expression of certain microRNAs (miRNAs) are elevated in murine polymicrobial periodontitis. The current paradigm is that miRNAs are generally involved in fine-tuning gene expression. However, our in vitro studies have demonstrated that miR- 146a is a dominant miRNA that can be up-regulated 30 to 200+ fold during lipopolysaccharide stimulation and, more importantly, that this increase is sustained for days. We have demonstrated that miR-146a is a key regulator in endotoxin-induced tolerance and cross-tolerance using a monocyte/macrophage-based system. Similarly, we have demonstrated that miR-132 is a dominant miRNA in peptidoglycan-stimulated monocytes and can induce cross-tolerance. In the current proposal, these dominant miRNAs will be examined using in vitro and in vivo systems to critically determine their role in our established murine model of periodontitis with 4 major well-characterized periodontal pathogens (Porphyromonas gingivalis, Treponema denticola, Tannerella forsythia, Fusobacterium nucleatum) and Streptococcus gordonii as early colonizer. The overall hypothesis is that these miRNAs are the dominant miRNAs regulating toll-like receptor (TLR)/IL-1-signaling pathways. Four Specific Aims are proposed. Specific Aim 1 will define the mechanistic role of these miRNAs, including mapping of target mRNAs, in primary human oral epithelial cells in reference to human monocytes. Specific Aim 2 will determine the expression kinetics of the dominant miRNA in mono- or time-sequential polymicrobial infection-induced periodontitis in mice and examine the relative effects of TLR2 and TLR4 using gene knockout mice. Specific Aim 3 will evaluate the relative contribution of these dominant miRNAs in this periodontitis model using specific miRNA knockout mice. Specific Aim 4 will investigate the association of these dominant miRNAs as biomarkers in gingival crevicular fluid and gingival tissues in chronic periodontitis and correlation with therapeutic outcomes. The long-term goal is to determine how extensively these dominant miRNAs can serve as functional biomarkers and they regulate innate immune response pathways contributing to periodontitis. In future studies, this mouse periodontitis model will become critical to help develop manipulation of these miRNA functions and/or the TLR pathway into novel therapeutics for periodontitis. Since innate immune response is known to play critical roles in many other diseases, our findings will likely be applicable to other chronic inflammatory and autoimmune diseases.

抽象的 仅在美国，牙周病就影响着数百万人，并已被证实为一种前兆 其他使人衰弱的全身性疾病，包括心血管疾病、阿尔茨海默病、类风湿病 关节炎和不良妊娠结局。我们的实验室已经证明某些 microRNA 的表达 (miRNA) 在小鼠多种微生物牙周炎中升高。目前的范式是 miRNA 通常涉及基因表达的微调。然而，我们的体外研究表明，miR- 146a 是一种显性 miRNA，在脂多糖刺激期间可上调 30 至 200 倍以上，并且， 更重要的是，这种增长持续了数天。我们已经证明 miR-146a 是一个关键 使用基于单核细胞/巨噬细胞的系统调节内毒素诱导的耐受性和交叉耐受性。 同样，我们已经证明 miR-132 是肽聚糖刺激的单核细胞中的主要 miRNA 并可诱导交叉耐受。在当前的提案中，将使用以下方法检查这些显性 miRNA： 体外和体内系统，以严格确定它们在我们建立的小鼠牙周炎模型中的作用，其中 4 主要的明确特征的牙周病原体（牙龈卟啉单胞菌、齿垢密螺旋体、坦纳菌 连翘、具核梭杆菌）和戈登链球菌作为早期定殖者。总体假设是 这些 miRNA 是调节 Toll 样受体 (TLR)/IL-1 信号通路的主要 miRNA。四 提出了具体目标。具体目标 1 将定义这些 miRNA 的机制作用，包括 参照人单核细胞，在原代人口腔上皮细胞中绘制目标 mRNA 的图谱。具体的 目标 2 将确定单一或时间序列多微生物中主要 miRNA 的表达动力学 小鼠感染诱发的牙周炎并使用基因敲除检查 TLR2 和 TLR4 的相对影响 老鼠。具体目标 3 将评估这些显性 miRNA 在此牙周炎模型中的相对贡献 使用特定的 miRNA 敲除小鼠。具体目标 4 将研究这些显性 miRNA 的关联 作为慢性牙周炎龈沟液和牙龈组织的生物标志物及其相关性 治疗结果。长期目标是确定这些显性 miRNA 的服务范围有多大 作为功​​能性生物标志物，它们调节导致牙周炎的先天免疫反应途径。在 在未来的研究中，这种小鼠牙周炎模型对于帮助开发这些 miRNA 的操作将变得至关重要 功能和/或 TLR 途径成为牙周炎的新疗法。由于先天免疫反应是 已知在许多其他疾病中发挥关键作用，我们的研究结果可能适用于其他慢性病 炎症和自身免疫性疾病。