Mechanistic probes to study the immune response in periodontal disease

研究牙周病免疫反应的机制探针

基本信息

批准号：
10189556
负责人：
Patrick M Woster
金额：
$ 40.47万
依托单位：
MEDICAL UNIVERSITY OF SOUTH CAROLINA
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-07-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10189556
关键词：
Actinobacillus actinomycetemcomitans Active Sites Adjuvant Adult Affect American Attenuated Automobile Driving Biochemical CRISPR/Cas technology Calvaria Chemicals Chromatin Clinical Collection Connective Tissue Diseases Crystallization Data Development Disease Disease Progression Docking Drug Design Drug Targeting Enzyme Kinetics Enzymes Epigenetic Process Family Fostering Genetic Transcription Goals Health Health Sciences Hematoxylin and Eosin Staining Method Histones Human Immune response In Vitro Infiltration Inflammation Inflammatory Inflammatory Response Interleukin-6 KDM1A gene Knock-out Knowledge Libraries Ligature Lipopolysaccharides Lysine Maps Maximum Tolerated Dose Measures Modeling Modification Mus Operative Surgical Procedures Oral health Osteoclasts Pathogenesis Pathologic Patients Periodontal Diseases Periodontic specialty Periodontitis Phenotype Play Population Process Production Proteins Research Role Sampling Source South Carolina Structure Surface Antigens System TNF gene Techniques Testing Therapeutic Therapeutic Agents Tissues Toxic effect analog base bone loss chromatin remodeling computational chemistry cytokine demethylation design experimental study high throughput screening histological stains histone demethylase immunoregulation in silico in vivo inflammatory bone loss inhibitor/antagonist lead optimization macrophage meetings microCT novel novel therapeutics osteoclastogenesis osteoimmunology periodontopathogen periopathogen pharmacophore prevent progression marker response scaffold side effect small molecule therapeutic target tool trimethyllysine

项目摘要

Abstract Periodontal diseases affect 42% of adult Americans and are characterized by bacterial-driven inflammatory bone loss. Traditional and emerging treatments for periodontitis management do not typically target the host immune response, which is the major source of tissue damage. The demethylation activity of the chromatin remodeling enzyme lysine-specific demethylase 1 (KDM1A) at the transcription activating mark histone 3 lysine 4 (H3K4) leads to a decrease in pro-inflammatory cytokine transcription. By contrast, the chromatin remodeling enzyme lysine specific demethylase 4B (KDM4B) specifically demethylates the transcription deactivating mark histone 3 trimethyllysine 9 (H3K9me3), leading to up regulated expression of pro-inflammatory cytokines (PICs). Interestingly, cross talk between these two enzymes leads to a balanced system wherein lysine 9 hypomethylation by KDM4B serves as a prerequisite to lysine 4 hypomethylation by KDM1A. The research plan outlined in this proposal will exploit this crosstalk for the design of new chemical probes for use in the study of the epigenetic basis for PD. The the central hypothesis of this study is that promotion of KDM1A activity by introduction of a specific KDM4B or KDM4E inhibitor will alleviate PD by reducing the expression of PICs in diseased tissue and reducing osteoclast formation. Inhibitors so identified will be useful as chemical probes to study the biochemical basis of inflammation and bone loss in PD. We will test this hypothesis through completion of the following Specific Aims: Specific Aim 1: We will use structure-based design techniques to discover novel inhibitors of KDM4B or 4E for use as chemical tools to elucidate the mechanism underlying inflammation and bone loss in PD; Specific Aim 2: We will define the cellular mechanism by which KDM4B/4E over expression contributes to periodontal inflammation and bone loss; Specific Aim 3: We will evaluate novel and known KDM4B/4E inhibitors for immunomodulatory activity in vivo using two models of PD. Our preliminary results demonstrate that KDM4B and 4E protein is more abundant in vivo in periodontally diseased connective tissue, and that inhibition of KDM4B results in significant decreases in PIC production and osteoclastogenesis in tissue pre-treated with a periopathogenic lipopolysaccharide. New and existing inhibitors will be used to further validate KDM4B as a therapeutic target in PD, and computational chemistry docking experiments paired with physical compound screens will be employed to correlate compound structure and changes in disease progression markers. Selected KDM4B inhibitors will be interrogated both in vitro and in vivo for efficacy and toxicity. This study will provide a more robust understanding of epigenetic mechanisms that play a significant role periodontal disease progression, validate KDM4B as a drug target for periodontitis, and result in development of chemical probes with therapeutic potential for local immunomodulatory adjuvant treatment of PD. This project will break down barriers between the fields of periodontics and drug design and will foster the development of knowledge on a critically needed aspect of translational oral health science.

抽象的牙周病影响 42% 的美国人，其特点是细菌驱动的炎症骨质流失。传统和新兴的牙周炎治疗方法通常不针对宿主免疫反应，这是组织损伤的主要来源。染色质的去甲基化活性转录激活标记组蛋白 3 赖氨酸上的重塑酶赖氨酸特异性去甲基化酶 1 (KDM1A) 4 (H3K4) 导致促炎细胞因子转录减少。相比之下，染色质重塑赖氨酸特异性去甲基化酶 4B (KDM4B) 特异性地使转录失活标记去甲基化组蛋白 3 三甲基赖氨酸 9 (H3K9me3)，导致促炎细胞因子表达上调（图片）。有趣的是，这两种酶之间的串扰导致了一个平衡系统，其中赖氨酸 9 KDM4B 引起的低甲基化是 KDM1A 引起的赖氨酸 4 低甲基化的先决条件。研究该提案中概述的计划将利用这种串扰来设计新的化学探针，用于 PD表观遗传学基础的研究。本研究的中心假设是 KDM1A 的促进作用通过引入特定的 KDM4B 或 KDM4E 抑制剂的活性将通过减少病变组织中的 PIC 并减少破骨细胞的形成。如此鉴定的抑制剂将可用作化学品探针研究帕金森病炎症和骨质流失的生化基础。我们将检验这个假设通过完成以下具体目标：具体目标 1：我们将使用基于结构的设计发现 KDM4B 或 4E 新型抑制剂的技术，用作阐明其机制的化学工具 PD 中潜在的炎症和骨质流失；具体目标 2：我们将定义细胞机制 KDM4B/4E 过度表达导致牙周炎症和骨质流失；具体目标 3：我们将使用两种 PD 模型评估新型和已知的 KDM4B/4E 抑制剂的体内免疫调节活性。我们的初步结果表明，KDM4B 和 4E 蛋白在牙周组织中的体内含量更加丰富患病结缔组织，抑制 KDM4B 会导致 PIC 产生显着减少，用病周脂多糖预处理的组织中的破骨细胞生成。新的和现有的抑制剂将用于进一步验证KDM4B作为PD的治疗靶点，以及计算化学对接与物理化合物筛选相结合的实验将用于关联化合物结构和疾病进展标志物的变化。选定的 KDM4B 抑制剂将在体外和体内进行研究体内功效和毒性。这项研究将为表观遗传机制提供更深入的理解在牙周病进展中发挥重要作用，验证 KDM4B 作为牙周炎的药物靶点，并导致开发具有局部免疫调节佐剂治疗潜力的化学探针治疗PD。该项目将打破牙周病学和药物设计领域之间的障碍将促进转化口腔健康科学急需方面知识的发展。