Inhibiting Periodontitis by Targeting Cathepsin K and Attenuating TLR Signaling

通过靶向组织蛋白酶 K 和减弱 TLR 信号传导抑制牙周炎

• 批准号: 9145992
• 负责人: YI-PING LI
• 金额: $ 8.23万
• 依托单位: UNIVERSITY OF ALABAMA AT BIRMINGHAM
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-01-01 至 2018-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9145992
• 关键词: Adult; Age; Alveolar Bone Loss; American; Animal Model; Arthritis; Atherosclerosis; Attenuated; Bacteria; Bone Resorption; Cardiovascular Diseases; Cells; Chronic; Data; Dendritic Cells; Dentists; Dependovirus; Diabetes Mellitus; Disease; Endodontics; Enzyme-Linked Immunosorbent Assay; Fibroblasts; Flow Cytometry; Fruit; Genes; Gingiva; Goals; Health; Host Defense; Human; Immune; Immune response; Immunity; Immunologist; In Vitro; Infection; Infiltration; Inflammation; Inflammation Mediators; Inflammatory; Injection of therapeutic agent; Interdisciplinary Study; Knowledge; Lead; Mediating; Modeling; Molecular; Mus; Oral; Osteitis; Osteoclasts; Pathogenesis; Peptide Hydrolases; Periodontal Diseases; Periodontitis; Prevention; Prevention strategy; Public Health; Rattus; Receptor Signaling; Reporting; Research; Rheumatoid Arthritis; Risk Factors; Role; Sampling; Scientist; Severities; Signal Transduction; Testing; Therapeutic; Tissues; Toll-like receptors; Tooth Loss; Wild Type Mouse; adaptive immunity; aged; alveolar bone; attenuation; base; bone loss; cathepsin K; combat; comparative; cytokine; design; effective therapy; gene therapy; improved; in vivo; insight; knockout gene; macrophage; mouse model; novel; novel therapeutic intervention; novel therapeutics; oral infection; pathogen; pre-clinical research; prevent; small hairpin RNA; stem; therapy development

DESCRIPTION (provided by applicant): The long-term objective of this proposal is to develop novel therapeutics for periodontitis by targeting cathepsin K (Ctsk) and attenuating Toll-like receptor (TLR) signaling. Periodontitis is one of the most common inflammatory diseases in humans that results in the destruction of periodontal tissues and alveolar bone, which ultimately leads to teeth loss. It is estimated that majority of adults over the age of 30 suffer from periodontal bone loss. Growing evidence suggests that chronic periodontal inflammation is an important risk factor for several pathological disorders including cardiovascular disease, diabetes, atherosclerosis and arthritis. Thus, periodontitis is a significant public health concern particularly to aged people. Consequently, there is still an urgent need for developing better treatments and preventative strategies that can dramatically reduce the inflammation, bone loss, and systemic ramifications of periodontitis. The current lack of highly effective therapies may be largely due to incomplete knowledge of the mechanism of periodontitis pathogenesis. Surprisingly, although TLRs are critical for host defense and inflammatory diseases, the role of TLRs in the pathogenesis of periodontitis remains largely unknown. We found that adeno-associated virus (AAV) Ctsk shRNAi (AAV-shRNA-Ctsk) mediated silencing prevents both bone loss and inflammation in a mouse model of endodontic disease. Moreover, it was reported that Ctsk is required for TLR9 signaling in a rat model of rheumatoid arthritis. Notably, our preliminary data showed that Cathepsin K gene knockout and AAV Ctsk shRNA (AAV-shRNA-Ctsk) mediated silencing dramatically prevents both bone loss and inflammation in a mouse model of periodontitis. Collectively, these studies strongly indicate that Ctsk is a major "osteoimmune gene" that can be targeted to control both inflammation and bone loss, and that Ctsk may be a key regulator of TLRs signaling. Based on our studies and those of others, we hypothesize that targeting Ctsk inhibits inflammation and bone loss caused by bacteria infection in periodontitis through attenuation of TLRs signaling. Three specific aims are proposed to test our hypothesis. We will define the functional role of Ctsk in the TLRs signaling-mediated immune response and bone resorption induced by periodontitis through comparative analysis of Ctsk-/-, Ctsk+/-, and Ctsk+/+ mice infected with periodontal pathogens in Aim 1. We will determine the therapeutic potential of AAV-shRNA-Ctsk as a means to reduce the progression and severity of periodontitis in vivo by attenuating TLRs signaling in Aim 2. We will characterize the mechanism by which Ctsk mediates TLRs signaling induced by pathogens and inflammatory mediators in a mouse periodontitis model using dendritic cells, fibroblasts and macrophages in Aim 3. A multidisciplinary research team (i.e. a molecular geneticist, dentist scientists, animal model experts and an immunologist) has been established to achieve the research goal. This study will not only improve our understanding mechanism of basic knowledge of the pathogenesis of periodontitis, but it will facilitate the design of novel therapeutic approaches fo this disease.

描述（由申请人提供）：本提案的长期目标是通过靶向组织蛋白酶K（Ctsk）和减弱Toll样受体（TLR）信号转导来开发牙周炎的新型治疗药物。牙周炎是人类最常见的炎症性疾病之一，其导致牙周组织和牙槽骨的破坏，最终导致牙齿脱落。据估计，大多数30岁以上的成年人患有牙周骨丢失。越来越多的证据表明，慢性牙周炎是包括心血管疾病、糖尿病、动脉粥样硬化和关节炎在内的多种病理性疾病的重要危险因素。因此，牙周炎是一个重要的公共卫生问题，特别是对老年人。因此，仍然迫切需要开发更好的治疗和预防策略，可以显着减少牙周炎的炎症，骨丢失和全身性后果。目前缺乏高效的治疗方法可能在很大程度上是由于对牙周炎发病机制的认识不完全。令人惊讶的是，尽管TLR对宿主防御和炎症性疾病至关重要，但TLR在牙周炎发病机制中的作用在很大程度上仍然未知。我们发现腺相关病毒（AAV）Ctsk shRNA（AAV-shRNA-Ctsk）介导的沉默在牙髓病小鼠模型中防止骨丢失和炎症。此外，据报道，在类风湿性关节炎的大鼠模型中，Ctsk是TLR 9信号传导所必需的。值得注意的是，我们的初步数据表明，组织蛋白酶K基因敲除和AAV Ctsk shRNA（AAV-shRNA-Ctsk）介导的沉默显着防止牙周炎小鼠模型中的骨丢失和炎症。总的来说，这些研究强烈表明，Ctsk是一个主要的“骨免疫基因”，可以有针对性地控制炎症和骨丢失，Ctsk可能是TLR信号传导的关键调节因子。基于我们和其他人的研究，我们假设靶向Ctsk通过减弱TLR信号传导抑制牙周炎中细菌感染引起的炎症和骨丢失。提出了三个具体目标来检验我们的假设。我们将通过比较分析目标1中感染牙周病原体的Ctsk-/-、Ctsk+/-和Ctsk+/+小鼠，确定Ctsk在TLR信号介导的免疫应答和牙周炎诱导的骨吸收中的功能作用。我们将确定AAV-shRNA-Ctsk作为通过减弱Aim 2中的TLR信号传导来减少体内牙周炎的进展和严重程度的手段的治疗潜力。我们将描述Ctsk介导TLR信号转导诱导的病原体和炎症介质在小鼠牙周炎模型中使用树突状细胞，成纤维细胞和巨噬细胞在目标3的机制。为了实现研究目标，成立了一个多学科研究小组（即分子遗传学家、牙医科学家、动物模型专家和免疫学家）。这项研究不仅将提高我们对牙周炎发病机制的基础知识的了解，而且将促进这种疾病新治疗方法的设计。