Treatment of Periodontitis by Homing M2 Macrophages

归巢 M2 巨噬细胞治疗牙周炎

• 批准号: 10197100
• 负责人: CHARLES SFEIR
• 金额: $ 47.01万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-07-01 至 2025-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10197100
• 关键词: Ablation; Address; Adoptive Transfer; Adult; Affect; Alveolar Bone Loss; Anti-Inflammatory Agents; Antibiotics; Bacterial Infections; Calculi; Cardiovascular Diseases; Cell Separation; Cells; Clinical; Clinical Treatment; Communities; Data; Debridement; Dental Calculus; Dental Plaque; Diabetes Mellitus; Disease; Disease Progression; Etiology; Genetic; Gingiva; Harvest; Helper-Inducer T-Lymphocyte; Homeostasis; Homing; Immune; Immune response; Immune system; Inflammation; Inflammatory; Inflammatory Response; Interleukin-10; Interleukin-4; Intervention; Knockout Mice; Ligands; Ligature; Light; Maxilla; Methods; Microbe; Microbial Biofilms; Modality; Modeling; Mouth Diseases; Mus; Nature; Nutrient; Pathogenesis; Patients; Periodicity; Periodontal Diseases; Periodontitis; Periodontium; Phase; Phenotype; Play; Population; Premature Birth; Preventive therapy; Process; Public Health; Rattus; Reporting; Research; Resolution; Role; Severities; Stains; Sum; System; Systemic disease; Technology; Testing; Therapeutic; Therapeutic Effect; Therapeutic Intervention; Tissue Harvesting; Tissues; Tooth Loss; Tooth structure; Translating; United States; Wild Type Mouse; aged; bone loss; chemokine; chronic inflammatory disease; cytokine; dental biofilm; dysbiosis; experimental study; feasibility testing; immunoregulation; in vivo; inflammatory bone loss; insight; macrophage; microCT; microbial; microbiota; monocyte; novel; novel therapeutic intervention; pathogen; prevent; recruit; repaired; response; single-cell RNA sequencing; standard care; standard of care; tumor-immune system interactions

Periodontitis (PD) is the second most common oral disease, affecting about half of adults (65 millions) in the United States. Periodontitis causes tooth loss and has been implicated in the pathogenesis of several serious systemic diseases including diabetes, cardiovascular diseases, and premature birth. Periodontitis is a chronic inflammatory disease, triggered by bacterial infection present in dental plaques and calculus; but actual disease manifestations are caused by host immune response to these pathogens. Current standard treatment is debridement of plaques and calculus to reduce the bacterial load; however, there are no therapies that address the immune component of PD. Thus, many research groups have started developing technologies to regulate the immune response and reduce inflammation. Our group has focused on recruiting and polarizing M2 macrophage as novel approaches for therapy of periodontitis. In our previous study, we successfully prevented alveolar bone loss by locally inducing M2 macrophages in mouse periodontitis models. M2 macrophages were polarized by injecting C-C motif chemokine 2 (CCL2) releasing PLGA microparticles (MPs) into periodontal tissues. The results indicated that inducing M2 macrophages by local delivery of CCL2 indeed reduces inflammation and bone loss in periodontitis. The proposed study will explore further the therapeutic potential of M2 macrophage induction by CCL2 as well as shed light on the mechanism underlying this process. We hypothesize that CCL2 plays a key role in maintaining periodontal immune homeostasis, and that its exogenous delivery, or genetic ablation, would modify the course of PD through periodontal breakdown, repair and microbiota dysbiosis. Macrophages play an important role in both the destructive and constructive phases of the inflammatory response by modulating their ability to polarize into either M1 (pro-inflammatory/pro-destructive) or M2 (anti-inflammatory/pro-reparative) macrophages. Our hypothesis is strongly supported by our recent in vivo data showing that the local delivery of CCL2 will induce differentiation of macrophages and monocytes to an M2 phenotype, thus leading to decreased periodontal inflammatory bone loss and initiation of periodontal repair. To test this hypothesis, we propose the following specific aims: 1) To test CCL2 MPs as an interventional and reparative periodontal therapy; 2) To study the periodontal phenotype, periodontal breakdown and repair in CCL2-knockout mice (CCL2-KO); and 3) To perform Single cell RNA sequencing (ScRNA-Seq) to acquire in- depth mechanistic data about the role of CCL2, its association, effect on and interactions with other immune cells. We anticipate that CCL2 released from PLGA MPs will inhibit further bone loss and promote repair in the mouse periodontitis model. We also expect that CCL2-KO mice and ScRNA-Seq analysis will provide data critical to understanding underlying mechanisms of CCL2’s immunoregulatory effects. In sum, we believe that M2 macrophage induction by CCL2 in inflamed periodontium could be a novel, promising strategy for treating PD.

牙周炎 (PD) 是第二常见的口腔疾病，影响着大约一半的成年人（6500 万人） 美国。牙周炎会导致牙齿脱落，并与多种严重疾病的发病机制有关 全身性疾病包括糖尿病、心血管疾病、早产等。牙周炎是一种慢性病 由牙菌斑和牙石中存在的细菌感染引发的炎症性疾病；但实际的疾病 表现是由宿主对这些病原体的免疫反应引起的。目前的标准治疗是 清创牙菌斑和牙石以减少细菌负荷；然而，没有治疗方法可以解决 PD 的免疫成分。因此，许多研究小组已经开始开发技术来调节 免疫反应并减少炎症。我们小组专注于招募和极化 M2 巨噬细胞作为治疗牙周炎的新方法。在我们之前的研究中，我们成功地阻止了 在小鼠牙周炎模型中局部诱导 M2 巨噬细胞导致牙槽骨丢失。 M2巨噬细胞是 通过将 C-C 基序趋化因子 2 (CCL2) 注射到牙周组织中释放 PLGA 微粒 (MP) 来极化 组织。结果表明，通过局部递送 CCL2 诱导 M2 巨噬细胞确实减少了 牙周炎中的炎症和骨质流失。拟议的研究将进一步探索其治疗潜力 CCL2 诱导 M2 巨噬细胞并揭示了这一过程的机制。我们 假设CCL2在维持牙周免疫稳态中发挥关键作用，并且其外源性 分娩或基因消融将通过牙周破坏、修复和治疗来改变 PD 的病程。 微生物群失调。巨噬细胞在破坏性和建设性阶段都发挥着重要作用 通过调节其极化为 M1（促炎症/促破坏）的能力来调节炎症反应 或 M2（抗炎/促修复）巨噬细胞。我们的假设得到了我们最近的研究的有力支持 体内数据显示，CCL2 的局部递送将诱导巨噬细胞和单核细胞分化为 M2 表型，从而减少牙周炎性骨丢失和牙周病的发生 维修。为了检验这一假设，我们提出以下具体目标：1）测试 CCL2 MP 作为干预措施 和修复性牙周治疗； 2）研究牙周表型、牙周破坏和修复 CCL2 基因敲除小鼠 (CCL2-KO)； 3) 进行单细胞 RNA 测序 (ScRNA-Seq) 以获取信息 关于 CCL2 的作用、其关联、对其他免疫的影响和相互作用的深度机制数据 细胞。我们预计 PLGA MP 释放的 CCL2 将抑制进一步的骨质流失并促进骨质修复 小鼠牙周炎模型。我们还预计 CCL2-KO 小鼠和 ScRNA-Seq 分析将提供关键数据 了解 CCL2 免疫调节作用的潜在机制。综上所述，我们认为M2 CCL2 在发炎的牙周组织中诱导巨噬细胞可能是治疗 PD 的一种新颖且有前景的策略。