Role of Notch-1/PLA2-IIA in oral dysbiosis, inflammation, and periodontal disease

Notch-1/PLA2-IIA 在口腔生态失调、炎症和牙周病中的作用

• 批准号: 10447008
• 负责人: Octavio Alberto Gonzalez
• 金额: $ 54.4万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2026-04-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10447008
• 关键词: Address; Adverse effects; Agreement; Animal Model; Antimicrobial Effect; Antimicrobial susceptibility; Apoptosis; Autophagocytosis; Bacteria; Cell Proliferation; Cell physiology; Cells; Clinical; Complex; Data; Development; Disease; Ecology; Enzymes; Epithelial; Epithelial Cells; Event; Exhibits; Firmicutes; Gene Expression; Gingiva; Health; Human; Immune; In Vitro; Inbred BALB C Mice; Infection; Inflammation; Inflammatory; Knowledge; Lead; Lesion; Mediating; Microbe; Modeling; Molecular; Mouse Strains; Mucosal Immunity; Mucous Membrane; Mus; Mutation; Natural Immunity; Oral; Oral health; PLA2G2A gene; Pathogenicity; Pathway interactions; Periodontal Diseases; Periodontitis; Phospholipase A2; Play; Porphyromonas gingivalis; Property; Proteobacteria; Public Health; Receptor Activation; Reporting; Research; Research Design; Resistance; Role; Surface; Testing; Time; Transgenic Mice; Variant; adaptive immunity; antimicrobial; base; bone loss; chemokine; chronic infection; chronic inflammatory disease; dysbiosis; gingipain; in vivo; innovation; macrophage; malignant mouth neoplasm; microbial; microbial community; microbiota; mouse model; nonhuman primate; notch protein; oral cavity epithelium; oral microbiome; oral pathogen; overexpression; pathogen; prevent; receptor; recruit; response

The mechanisms by which mucosal responses to the microbiota in the presence of specific pathogenic bacterial species (e.g. P. gingivalis-Pg), could abrogate the host-microbe symbiotic relationship leading to dysbiosis and inflammatory disease remain not fully understood. Our pioneering R21 studies demonstrated that Pg-induced phospholipase A2 group IIA (PLA2-IIA), in a mechanism that involves activation of Notch-1 receptor (N1R) by Pg gingipains, modulates the antimicrobial properties of oral epithelial cells (OECs). This is consistent with in vivo and clinical evidence supporting the potent (ng/mL) antimicrobial effects of PLA2-IIA. Noteworthy, oral bacterial species exhibited differential antimicrobial susceptibility to PLA2-IIA. Moreover, gingival PLA2-IIA expression and N1R activation were elevated during initiation and progression of periodontal disease, which was concurrent with oral dysbiosis in non-human primates. In agreement with these observations, new preliminary studies using mice models demonstrated increased gingival PLA2-IIA expression and N1R activation early after Pg infection. Remarkably, the oral microbiome of transgenic mice overexpressing PLA2-IIA (PLA2-IIA-Tg) exhibited significant differences in the abundance of bacterial species (decreased Firmicutes and increased Proteobacteria) compared to their wild type co-caged littermates. Oral dysbiosis in PLA2-IIA-Tg mice was associated with changes in the gingival expression of genes involved in intracellular sensing (NOD2), classical antimicrobial factors (S100a8/S100a9, hBD1), and M1/M2 macrophage chemokines. N1R is a critical modulator of mucosal immunity; therefore, the ability of Pg to activate N1R in OECs is an exciting and innovative observation. Preliminary data suggested that other OEC functions/responses (bacterial sensing, apoptosis, and autophagy) could involve Pg-induced N1R activation. Based on this evidence, we hypothesize that activation of Notch-1/PLA2-IIA pathway in gingival epithelial surfaces would be an innovative and plausible mechanism by which pathogens such as Pg could specifically enhance oral dysbiosis, inflammation and periodontal disease. To test this hypothesis, we propose three specific aims: (i) To determine the role of PLA2-IIA in Pg-induced oral dysbiosis, inflammation and periodontal disease, (ii) To determine the role of epithelial Notch-1 in Pg-induced PLA2-IIA and periodontitis, and (iii) To identify and validate oral epithelial innate responses modulated by Pg through Notch-1 activation. To address these knowledge gaps, we will use the Pg-induced periodontitis oral gavage model in mouse strains with or without a natural PLA2-IIA mutation, PLA2-IIA-Tg mice with their WT littermates, epithelial-specific N1R deficient mice, as well as OEC cultures to validate the role of this pathway in Pg-induced oral dysbiosis and periodontal disease. These findings will enable a better understanding of the cellular and molecular mechanisms that are specifically modulated by pathogens to alter microbial communities at mucosal surfaces as well as contribute to the evidence for future research designed to determine the role of N1R in oral health and other diseases (e.g., oral cancer).

在特定致病细菌存在下，粘膜对微生物群的反应的机制 物种（例如牙龈疟原虫）可以消除宿主 - 菌共生关系，导致营养不良和 炎症性疾病尚未完全了解。我们的开创性R21研究表明，PG诱导 磷脂酶A2 IIA（PLA2-IIA），涉及通过Notch-1受体（N1R）激活Notch-1受体（N1R）的机制 PG牙龈，调节口腔上皮细胞（OEC）的抗菌特性。这与IN一致 体内和临床证据支持PLA2-IIA的有效（NG/ML）抗菌作用。值得注意的，口头 细菌物种表现出对PLA2-IIA的差异抗菌敏感性。此外，牙龈PLA2-IIA 在牙周疾病的启动和进展过程中，表达和N1R激活升高，这是 与非人类灵长类动物的口腔失调同时出现。与这些观察结果一致，新的初步 使用小鼠模型的研究表明，牙龈PLA2-IIA的表达和N1R激活较早。 PG感染。值得注意的是，过表达PLA2-IIA（PLA2-IIA-TG）的转基因小鼠的口服微生物组 在细菌种类的丰度上表现出显着差异（较浓汤和增加 与野生型共同式同窝窝板相比，蛋白质细菌）。 PLA2-IIA-TG小鼠的口腔营养不良是 与涉及细胞内感测（NOD2）的基因的牙龈表达的变化，经典 抗菌因子（S100A8/S100A9，HBD1）和M1/M2巨噬细胞趋化因子。 N1R是关键调节器 粘膜免疫；因此，PG在OEC中激活N1R的能力是令人兴奋且创新的 观察。初步数据表明其他OEC功能/反应（细菌感应，凋亡和 自噬）可能涉及PG诱导的N1R激活。基于这些证据，我们假设激活 牙龈上皮表面中的Notch-1/pla2-IIA途径将是一种创新和合理的机制 哪些病原体（例如PG）可以特异性增强口腔失调，炎症和牙周疾病。到 检验该假设，我们提出了三个具体目的：（i）确定PLA2-IIA在PG诱导的 口腔营养不良，炎症和牙周疾病，（ii）确定上皮的作用 PG诱导的PLA2-IIA和牙周炎中的Notch-1，以及（iii）以识别和验证口服 PG通过Notch-1激活调节了上皮先天反应。解决这些 知识差距，我们将在有或没有A 天然PLA2-IIA突变，pla2-iia-tg小鼠与wt同窝仔，上皮特异性N1R缺乏小鼠， 以及OEC培养物来验证该途径在PG诱导的口腔失调和牙周疾病中的作用。 这些发现将使对细胞和分子机制有更好的了解，这些机制是专门的 病原体调节以改变粘膜表面的微生物群落，并有助于证据 对于未来的研究，旨在确定N1R在口腔健康和其他疾病（例如口腔癌）中的作用。