Mechanistic probes to study the immune response in periodontal disease

研究牙周病免疫反应的机制探针

• 批准号: 10600100
• 负责人: Yuri Karl Peterson
• 金额: $ 40.61万
• 依托单位: MEDICAL UNIVERSITY OF SOUTH CAROLINA
• 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 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10600100
• 关键词: Actinobacillus actinomycetemcomitans; Active Sites; Adjuvant Therapy; Adult; Affect; American; Attenuated; Automobile Driving; Biochemical; CRISPR/Cas technology; Calvaria; Chemicals; Chromatin; Clinical; Collection; Connective Tissue Diseases; 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; Inflammation; Inflammatory; Inflammatory Infiltrate; Inflammatory Response; Interleukin-6; KDM1A gene; Knock-out; Knowledge; Libraries; Ligature; Lipopolysaccharides; Lysine; Macrophage; 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; 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; lead optimization; meetings; microCT; novel; novel therapeutics; osteoclastogenesis; osteoimmunology; overexpression; periodontopathogen; periopathogen; pharmacophore; prevent; progression marker; response; scaffold; side effect; small molecule; therapeutic target; tool; trimethyllysine

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 牙周疾病影响42％的成年美国人，其特征是细菌驱动的炎症 骨丢失。牙周炎管理的传统和新兴治疗方法通常不会针对宿主 免疫反应，这是组织损伤的主要来源。染色质的脱甲基活性 在转录激活标记组蛋白3赖氨酸的重塑酶赖氨酸特异性脱甲基酶1（KDM1A） 4（H3K4）导致促炎细胞因子转录的减少。相比之下，染色质重塑 酶赖氨酸特异性脱甲基酶4B（KDM4B）特异性脱甲基脱​​甲基二甲基二甲基脱甲基脱甲基脱甲基脱甲基酶 组蛋白3三甲基透析9（H3K9ME3），导致促炎细胞因子的调节表达 （图片）。有趣的是，这两种酶之间的串扰导致赖氨酸9的平衡系统9 KDM4B的低甲基化是通过KDM1A赖氨酸4甲基化的先决条件。研究 该提案中概述的计划将利用此串扰，以设计新的化学探针供该化学探针 PD的表观遗传基础的研究。这项研究的中心假设是促进KDM1A 通过引入特定的KDM4B或KDM4E抑制剂的活动将通过降低表达来减轻PD 病态组织中的照片并减少破骨细胞的形成。如此确定的抑制剂将是化学物质的 研究PD炎症和骨质流失的生化基础的探针。我们将检验这个假设 通过完成以下特定目的：特定目标1：我们将使用基于结构的设计 发现KDM4B或4E的新型抑制剂的技术用作化学工具以阐明机制 PD的潜在炎症和骨质流失；特定目的2：我们将定义细胞机制 表达上的KDM4B/4E有助于牙周炎症和骨质流失；特定目标3：我们将 使用两种PD模型评估新型和已知的KDM4B/4E抑制剂，用于体内免疫调节活性。 我们的初步结果表明，KDM4B和4E蛋白在体内更丰富 患病的结缔组织以及对KDM4B的抑制会导致PIC产生和 用毛细胞病的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中的治疗靶标，计算化学对接 将采用与物理化合物筛选配对的实验，以相关联合结构和 疾病进展标记的变化。选定的KDM4B抑制剂将在体外和在 体内功效和毒性。这项研究将为表观遗传机制提供更强烈的理解 那起着牙周疾病进展的重要作用，验证KDM4B是牙周炎的药物靶标， 并导致具有治疗潜力的化学探针的开发 Pd的处理。该项目将打破牙周和药物设计领域之间的障碍以及 将促进对翻译口腔健康科学急需方面的知识发展。