Neurobiological control of periodontal homeostasis through microRNA, TGF-beta, and Wnt signaling

通过 microRNA、TGF-β 和 Wnt 信号传导对牙周稳态的神经生物学控制

• 批准号: 9756189
• 负责人: Tom Diekwisch
• 金额: $ 15.39万
• 依托单位: TEXAS A&M UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-04 至 2020-02-18
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9756189
• 关键词: Affect; Animal Model; Animals; Ankylosis; Bioinformatics; Biological Assay; Bone Regeneration; Bone remodeling; Cicatrix; Collagen; Cytoskeletal Modeling; Cytoskeletal Proteins; Data; Data Set; Defect; Dentition; Diagnostic radiologic examination; Down-Regulation; Emotional; Engineering; Epithelial; Epithelial Cells; G-Protein-Coupled Receptors; Galanin; Ganglia; Gene Expression; Gene Expression Profiling; Goals; Health; Height; Homeostasis; Hybrids; In Vitro; Interruption; Jaw; Laboratories; Messenger RNA; MicroRNAs; Modeling; Nanosphere; Nerve; Nerve Fibers; Neurobiology; Neuropeptides; Operative Surgical Procedures; Osteoblasts; Osteoclasts; Osteocytes; Osteogenesis; Outcome; Pathway interactions; Patients; Peptides; Periodontal Diseases; Periodontal Ligament; Periodontal Pocket; Periodontitis; Periodontium; Phenotype; Prevention; Proteins; Rattus; Research; Rest; Reverse Transcriptase Polymerase Chain Reaction; Roentgen Rays; Role; Sensory; Signal Transduction; Signaling Molecule; Testing; Tissue Engineering; Tissues; Tooth Ankylosis; Tooth structure; Transforming Growth Factor beta; Trigeminal nerve structure; Up-Regulation; WNT Signaling Pathway; Walkers; alveolar bone; base; bone; clinically significant; craniofacial bone; design; glycogen synthase kinase 3 beta; inferior alveolar nerve; inhibitor/antagonist; innovation; mandibular nerve; mineralization; nerve supply; nerve transection; osteoclastogenesis; osteogenic; prevent; progenitor; response; sample fixation; skeletogenesis; soft tissue; spatial relationship; trend

The periodontium is a richly innervated tissue that undergoes continuous modelling and remodeling by alveolar bone osteoblasts, osteocytes, and osteoclasts. Studies from our laboratory have demonstrated a close spatial relationship between trigeminal nerve ganglia and the Epithelial Rests of Malassez (ERM), an epithelial cell network residing within the non-mineralized periodontal ligament. Inferior alveolar nerve (IAN) transection studies resulting in dento-alveolar ankylosis and a reduction in ERM have confirmed the essential role of sensory innervation for periodontal homeostasis. For the present application, we have established the IAN transection model in our laboratory and provided radiographic evidence for enhanced mineralization and ankylosis in the periodontal region of rat molars. Gene expression profiling comparing IAN transected and control tissues demonstrated an unexpected 28-fold significant increase in galanin (GAL) and a more than two-fold decrease in the TGF-β signaling molecules Smad2, Smad3 and Tgf-β1, and the Wnt inhibitors Dkk1, Dkk2 and Gsk-3β. In the same IAN transection group, microRNA miR-92b expression was more than two-fold upregulated, as verified via miRNA profiling and RT-PCR. MiR-92b upregulation after IAN transection in conjunction with bioinformatics data implicating Wnt and Tgf-β as possible miR-92b targets prompted us to speculate that GAL affects its skeletogenic downstream effects through miR-92b. In vitro studies revealed that GAL treatment promoted osteogenic differentiation of PDL progenitors and increased mineralization, while reducing osteoclastogenesis of BMMCs. Block of RhoA or application of GAL antagonist affected PDL cytoskeletal organization and gene expression, indicating that GAL functions through G protein coupled receptors. IWhen applied to periodontal pockets of animals suffering from periodontitis, GAL tissue engineering constructs accomplished a 20% increase in alveolar bone levels compared to controls, resulting in a clinically significant increase in alveolar bone height. Based on this promising new set of data we have designed a research plan to define the role of GAL in response to IAN transection, determine the mechanism underlying its effect on skeletogenesis, and exploit its applicability for bone regeneration and the prevention of periodontal ankylosis. The overall goal of our research plan is to test the hypothesis that periodontal nerves affect alveolar bone homeostasis through a GAL–GPCR–miR-92b–TGF-β/Wnt regulatory loop and that application of GAL neurosecretory peptides will stimulate Wnt signaling and new bone formation. We anticipate that the outcomes of our study will lead to innovative neuropeptide based/engineering hybrid approaches that will restore periodontal health in millions of patients and prevent the emotional and functional scars associated with lost teeth and incomplete dentitions.

牙周组织是一种神经支配丰富的组织，通过不断的建模和重塑 牙槽骨成骨细胞、骨细胞和破骨细胞。我们实验室的研究表明 三叉神经节和马拉塞斯上皮残余（ERM）之间密切​​的空间关系， 存在于非矿化牙周膜内的上皮细胞网络。下牙槽神经 (IAN) 导致牙槽骨强直和 ERM 降低的横断研究已证实 感觉神经支配对牙周稳态的重要作用。对于本应用程序，我们有 在我们的实验室建立了IAN横断模型，并为增强的射线照相证据提供了证据。 大鼠磨牙牙周区的矿化和强直。基因表达谱比较 IAN 横切组织和对照组织显示甘丙肽意外显着增加 28 倍 (GAL) 以及 TGF-β 信号分子 Smad2、Smad3 和 Tgf-β1 减少两倍以上， 以及 Wnt 抑制剂 Dkk1、Dkk2 和 Gsk-3β。在同一 IAN 横切组中，microRNA miR-92b 通过 miRNA 分析和 RT-PCR 验证，表达上调了两倍以上。 miR-92b IAN 横断后的上调结合生物信息学数据表明 Wnt 和 Tgf-β 为 可能的 miR-92b 靶标促使我们推测 GAL 影响其成骨下游效应 通过 miR-92b。体外研究表明 GAL 治疗促进 PDL 成骨分化 祖细胞和增加矿化，同时减少 BMMC 的破骨细胞生成。 RhoA 块或 GAL拮抗剂的应用影响了PDL细胞骨架组织和基因表达，表明 GAL 通过 G 蛋白偶联受体发挥作用。 I当应用于动物牙周袋时 患有牙周炎，GAL 组织工程结构使牙槽骨增加了 20% 与对照组相比，骨水平显着增加，导致牙槽骨高度在临床上显着增加。基于 根据这组有希望的新数据，我们设计了一项研究计划来定义 GAL 在响应中的作用 IAN 横切，确定其对骨骼发生影响的机制，并利用其 适用于骨再生和预防牙周强直。我们的总体目标 研究计划是检验牙周神经影响牙槽骨稳态的假设 通过 GAL–GPCR–miR-92b–TGF-β/Wnt 调节环以及 GAL 的应用 神经分泌肽将刺激 Wnt 信号传导和新骨形成。我们预计 我们的研究成果将带来基于神经肽/工程混合的创新方法 恢复数百万患者的牙周健康并预防相关的情感和功能性疤痕 牙齿缺失和牙列不完整。