Characterization of Treponema Denticola-Mediated Interactions With Periodontal Ligament Cells Leading to Persistent Host Cell/Tissue Destruction

齿垢密螺旋体介导的与牙周膜细胞相互作用导致宿主细胞/组织持续破坏的特征

• 批准号: 10217098
• 负责人: Erin Trent Malone
• 金额: $ 5.18万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-05 至 2023-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10217098
• 关键词: Address; Adult; Adverse effects; American; Bacteremia; Bacteria; Behavior; Cell Communication; Cell Culture Techniques; Cell Differentiation process; Cell membrane; Cell physiology; Cells; Characteristics; Chronic; Clinical; Complex; Data; Development; Disease; Dose; Elements; Enzymes; Epithelial; Epithelial Cells; Event; Exhibits; Extracellular Matrix Degradation; Food Preservatives; Gelatinase A; Genetic Transcription; Gram-Negative Bacteria; Gram-Positive Bacteria; Homeostasis; Human; Immune; Immune Evasion; Immune response; In Vitro; Investigation; Lead; Lipoproteins; Literature; MMP2 gene; Matrix Metalloproteinases; Mediating; Membrane Proteins; Metalloproteases; Molecular; Natural regeneration; Nisin; Oral; Oral cavity; Pathogenesis; Pathogenicity; Peptide Hydrolases; Periodontal Diseases; Periodontal Ligament; Periodontitis; Periodontium; Phenotype; Physiological; Prevalence; Probiotics; Process; Proteins; Publishing; Reporting; Role; Serine Protease; Source; Systemic disease; Testing; Therapeutic; Therapeutic Agents; Tissues; Transcription Process; Translating; Treponema denticola; Work; aged; antimicrobial; bactericide; bacteriocin; catalyst; cell behavior; combat; commensal bacteria; dentilisin; dysbiosis; extracellular; member; microbial; microbiome; migration; mutant; oral bacteria; oral biofilm; oral spirochetes; pathogenic bacteria; pathogenic microbe; protein complex; repaired; soft tissue; success

Project Summary/Abstract Periodontal disease is characterized by destruction of the hard and soft tissues that comprise the periodontium. This destruction translates to a degradation of the extracellular matrices (ECM), which is mediated by bacterial proteases, host-derived matrix metalloproteinases (MMPs), and other proteases released by host tissues and immune cells. Furthermore, pathogenic microbes are a catalyst for periodontal disease pathogenesis since the development of a dysbiosis of the microbiome enhances a persistent immune response. These bacterial pathogens interact with host tissue and thereby trigger adverse cellular functions, including a heightened immune response, tissue destruction, and tissue migration. One implicated bacteria, the oral spirochete, Treponema denticola (T. denticola or Td), is highly associated with periodontal disease (5). We’ve demonstrated that a Td outer membrane protein complex called dentilisin, contributes to the chronic activation of pro-MMP-2 in periodontal ligament (PDL) cells (26,27). Dentilisin exposure also triggers increased expression levels of activators and effectors of active MMP-2 in PDL cells. Despite these advances, there is no mechanism known for dentilisin-induced MMP-2 activation or PDL cytopathic behaviors leading to disease, or potential treatment. One possible therapeutic to combat these interactions is nisin. Nisin is a naturally occurring bacteriocin, widely used food preservative, and probiotic agent naturally made by bacterial species similar to those in the gut and oral cavity. Our lab demonstrated that nisin is an effective anti-microbial against key periodontopathic bacteria at low concentrations, including Td, with the potential to disrupt oral biofilm formation in vitro (11). Importantly, although most nisin related literature documents its role against gram-positive bacteria, our published work (11) highlights its efficacy against gram-negative bacteria. Furthermore, nisin at low concentrations is also more effective against certain oral pathogenic versus commensal bacteria; making it a potentially useful and selective agent for oral applications. Proper clearance of Td may provide a more symbiotic microbiome and “reset” homeostatic conditions needed for repair, remodeling or regeneration of the periodontium. Thus, the main hypotheses of this proposal are that T. denticola interactions with PDL cells mediate adverse effects on homeostasis and cellular functions leading to a compromised cellular phenotype. Additionally, nisin is a potential therapeutic for abrogating these effects. This hypothesis will be tested in the following specific aims: 1) Determine the mechanism of dentilisin-induced activation of pro- MMP-2 in PDL cells; 2) Characterize the effects of T. denticola(Td) challenge on PDL cell processes and differentiation; 3) Determine whether sub-antimicrobial doses of nisin can abrogate Td/dentilisin-mediated activation of MMP-2 in PDL cells. Success of this proposal will provide molecular evidence for a mechanism of Td-mediated adverse cytopathic effects leading to characteristics of severe periodontitis; as well as evidence on nisin as a potential therapeutic for reducing or reversing Td-mediated periodontopathic processes in vitro.

项目概要/摘要 牙周病的特点是构成牙周组织的硬组织和软组织遭到破坏。 牙周组织。这种破坏转化为细胞外基质（ECM）的降解，即 由细菌蛋白酶、宿主来源的基质金属蛋白酶 (MMP) 和其他蛋白酶介导 由宿主组织和免疫细胞释放。此外，病原微生物是牙周病的催化剂。 疾病发病机制，因为微生物群失调的发展增强了持续的免疫 回复。这些细菌病原体与宿主组织相互作用，从而引发不利的细胞功能， 包括增强的免疫反应、组织破坏和组织迁移。一种涉及细菌， 口腔螺旋体、齿垢密螺旋体（T. denticola 或 Td）与牙周病高度相关 (5)。 我们已经证明，一种称为牙菌素 (dentilisin) 的 Td 外膜蛋白复合物会导致慢性 牙周膜 (PDL) 细胞中 pro-MMP-2 的激活 (26,27)。接触牙本质也会引发增加 PDL 细胞中活性 MMP-2 激活剂和效应子的表达水平。尽管取得了这些进步，但还没有 已知牙菌素诱导的 MMP-2 激活或导致疾病的 PDL 细胞病变行为的机制，或 潜在的治疗。对抗这些相互作用的一种可能的治疗方法是乳链菌肽。 Nisin 是一种天然存在的 细菌素，广泛使用的食品防腐剂，以及由类似于细菌种类天然产生的益生菌剂 那些在肠道和口腔中的。我们的实验室证明乳链菌肽是一种有效的抗微生物剂，可对抗关键微生物 低浓度的牙周病细菌，包括 Td，有可能破坏口腔生物膜的形成 体外 (11)。重要的是，尽管大多数乳链菌肽相关文献都记载了其对抗革兰氏阳性菌的作用 细菌，我们发表的工作 (11) 强调了它对革兰氏阴性细菌的功效。此外，乳链菌肽 与共生细菌相比，低浓度对某些口腔致病菌也更有效；制作它 一种用于口服应用的潜在有用的选择性试剂。 Td 的适当清除可能会提供更多 共生微生物组和“重置”修复、重塑或再生所需的稳态条件 牙周组织。因此，该提案的主要假设是 T. denticola 与 PDL 的相互作用 细胞介导对稳态和细胞功能的不利影响，导致细胞受损 表型。此外，乳链菌肽是消除这些影响的潜在治疗方法。这个假设将 在以下具体目标中进行测试： 1) 确定牙菌素诱导的前体激活的机制 PDL 细胞中的 MMP-2； 2) 表征 T. denticola (Td) 攻击对 PDL 细胞过程的影响和 差异化； 3) 确定亚抗菌剂量的乳链菌肽是否可以消除 Td/dentilisin 介导的作用 PDL 细胞中 MMP-2 的激活。该提案的成功将为以下机制提供分子证据： Td 介导的不良细胞病变效应导致严重牙周炎的特征；以及证据 乳链菌肽作为一种潜在的治疗方法，可在体外减少或逆转 Td 介导的牙周病过程。