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

基本信息

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
项目类别：
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

项目摘要

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).

在特定病原菌存在的情况下，粘膜对微生物群的反应机制种（例如 P. gingivalis-Pg），可能会破坏宿主与微生物的共生关系，导致菌群失调和炎症性疾病仍未完全了解。我们开创性的 R21 研究表明，Pg 诱导磷脂酶 A2 IIA 组 (PLA2-IIA)，其机制涉及通过激活 Notch-1 受体 (N1R) Pg gingipains 可调节口腔上皮细胞 (OEC) 的抗菌特性。这与在体内和临床证据支持 PLA2-IIA 的有效 (ng/mL) 抗菌作用。值得注意的是，口头细菌种类对 PLA2-IIA 表现出不同的抗菌敏感性。此外，牙龈PLA2-IIA 在牙周病的发生和进展过程中，N1R 的表达和激活水平升高，与非人类灵长类动物的口腔生态失调同时发生。与这些观察一致，新的初步使用小鼠模型的研究表明，术后早期牙龈 PLA2-IIA 表达和 N1R 激活增加 PG感染。值得注意的是，转基因小鼠的口腔微生物组过度表达 PLA2-IIA (PLA2-IIA-Tg) 细菌种类丰度存在显着差异（厚壁菌门减少，而细菌种类增加）变形菌）与其野生型同笼同窝动物相比。 PLA2-IIA-Tg 小鼠的口腔生态失调与细胞内传感 (NOD2) 相关基因的牙龈表达变化相关，经典抗菌因子（S100a8/S100a9、hBD1）和 M1/M2 巨噬细胞趋化因子。 N1R 是一个关键的调制器粘膜免疫；因此，Pg 激活 OEC 中 N1R 的能力是令人兴奋和创新的观察。初步数据表明其他 OEC 功能/反应（细菌感应、细胞凋亡和自噬）可能涉及 Pg 诱导的 N1R 激活。基于这一证据，我们假设激活牙龈上皮表面的 Notch-1/PLA2-IIA 通路将是一种创新且合理的机制 Pg等病原体可以特异性增强口腔生态失调、炎症和牙周病。到为了检验这一假设，我们提出了三个具体目标：(i) 确定 PLA2-IIA 在 Pg 诱导中的作用口腔生态失调、炎症和牙周病，(ii) 确定上皮细胞的作用 Pg 诱导的 PLA2-IIA 和牙周炎中的 Notch-1，以及 (iii) 识别和验证口腔 Pg 通过 Notch-1 激活调节上皮先天反应。为了解决这些由于知识差距，我们将在有或没有牙周炎的小鼠品系中使用 Pg 诱导的牙周炎口服灌胃模型。天然 PLA2-IIA 突变、PLA2-IIA-Tg 小鼠及其 WT 同窝小鼠、上皮特异性 N1R 缺陷小鼠、以及 OEC 培养物，以验证该途径在 Pg 诱导的口腔生态失调和牙周病中的作用。这些发现将有助于更好地理解特定的细胞和分子机制。受病原体调节以改变粘膜表面的微生物群落并有助于证据用于未来旨在确定 N1R 在口腔健康和其他疾病（例如口腔癌）中的作用的研究。