Mechanosensing of osteoclasts in periodontitis.

牙周炎中破骨细胞的机械传感。

• 批准号: 10752476
• 负责人: Satoru Shindo
• 金额: $ 8.41万
• 依托单位: NOVA SOUTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-04 至 2026-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10752476
• 关键词: ANXA5 gene; Actinobacillus actinomycetemcomitans; Actins; Address; Affect; Alkaline Phosphatase; Applications Grants; Awards and Prizes; Binding; Binding Proteins; Biology; Biomedical Research; Bone Regeneration; Bone Resorption; Bone remodeling; Cells; Chemicals; Coculture Techniques; Consultations; Core Facility; Coupling; Cytochalasin D; Cytoskeleton; Data; Data Set; Development; Devices; Down-Regulation; Ensure; Enzyme-Linked Immunosorbent Assay; Equipment; Florida; Forsythia; Foundations; Future; Goals; Grant; Image; Immune; In Vitro; Inflammatory Response; Insulin-Like Growth Factor I; Interferometry; Laboratories; Laboratory Personnel; Ligature; Link; Lipids; Mechanical Stimulation; Mechanical Stress; Mechanics; Mediating; Mentors; Methods; Microbiology; Microscopy; Molecular; Monitor; Motor; Mus; Mycotoxins; Myosin Type II; Nobel Prize; Oral Administration; Osteoblasts; Osteoclasts; Osteocytes; Osteoporotic; Outcome; Pathogenicity; Periodontitis; Periodontium; Phenotype; Phosphatidylserines; Piezo 1 ion channel; Piezo 2 ion channel; Porphyromonas gingivalis; Postdoctoral Fellow; Process; Production; Proteins; Protocols documentation; Raman Spectrum Analysis; Reporting; Research; Research Personnel; Research Project Grants; Risk Factors; Role; Science; Signal Transduction; Small Interfering RNA; System; Systemic disease; TNFSF11 gene; Techniques; Testing; Tissues; Tooth structure; Training; Transferase; Virulence Factors; Virulent; Writing; alveolar bone; bone; bone loss; career development; cathepsin K; complex data; dihydroceramide; experience; gain of function; immunopathology; improved; in vivo imaging; inhibitor; lipidomics; loss of function; mechanotransduction; microCT; morphometry; mouse model; neutralizing antibody; non-muscle myosin; novel; novel therapeutic intervention; osteoimmunology; periodontopathogen; precursor cell; programs; regeneration potential; response; shear stress; skills

Project Summary/Abstract Host immune inflammatory response to periodontal pathogens, especially, Porphyromonas gingivalis (Pg), is engaged in the destruction of tooth-supporting tissues in periodontitis. Among several virulent factors produced by Pg, a unique pathogenic lipid, phosphoglycerol dihydroceramide (PGDHC), promotes RANKL-induced OC- genesis by acting on non-muscle myosin II-A (Myh9), while inhibiting the production of OB-genesis factor, IGF- 1, from OCs. In our preliminary results, wild-type Pg cells and purified PGDHC, but not serine palmitoyl transferase (SPT) KO Pg cells (deficient in PGDHC production), abrogated mechanosensitive Piezo1-mediated downregulation of OC-genesis. The data suggested that PGDHC may represent a unique virulence factor that causes dysregulation of Piezo1-mechanosnsing system in osteoclasts which leads to retarded bone regeneration. Based on these lines of evidence and preliminary results, it is hypothesized that mechanosensitive Piezo1 elicits a cell signal in OCs that induces the production of IGF-1 and suppress OC- genesis, while the PGDHC, by its binding to Myh9, blocks the mechanosensing function of Piezo1 and, hence, diminishing the bone regenerative potential of alveolar bone affected by periodontitis. This hypothesis will be tested by the following two Specific Aims: Aim 1) To elucidate the molecular mechanism underlying PGDHC- mediated inhibition of Piezo1 channel expressed on OCs in vitro. Aim 2) To determine the effect of PGDHC on the Piezo1 expressed by OCs in a mouse model of ligature and Pg-induced periodontitis. Major four training goals for the candidate include: 1) Gain and improve expertise in advanced science methods associated with osteoimmunology and periodontology, 2) Improve skills in management and analyses of complex datasets, 3) Acquire novel image acquisition and analysis techniques to link bacterial virulent factor and Piezo1 expressed on osteoclasts, using Raman spectroscopy, in vivo imaging, micro-CT and biolayer interferometry, and 4) Develop skills required to lead a research program, including grant writing, management of laboratory personnel, project development and execution. Sponsor, Dr. Kawai, and co-sponsor, Dr. Han, who are experienced in the proposed study addressing the periodontal osteoimmunology and immunopathology as well as mentoring of postdoctoral fellows for their career development in the academic research fields, will be committed to support the candidate’s training, along with consultation committees composed of 5 expert researchers in the fields of lipidomics, bone biology, mechanosensory system, microbiology, and Raman spectroscopy, respectively. All required equipment, devices, protocols and materials are available in the laboratories of sponsors and consultants. The candidate is given a desk and laboratory space in Center for Collaborative Research (CCR) which is the largest biomedical research building in South Florida, equipped with a variety of cutting-edge equipment in the core facilities, all of which are available for the candidate’s research project.

项目概要/摘要 宿主对牙周病原体，尤其是牙龈卟啉单胞菌（Pg）的免疫炎症反应 参与牙周炎中牙齿支持组织的破坏。 Pg 是一种独特的致病脂质，磷酸甘油二氢神经酰胺 (PGDHC)，可促进 RANKL 诱导的 OC- 通过作用于非肌肉肌球蛋白 II-A (Myh9)，同时抑制 OB 发生因子、IGF- 的产生， 1，来自 OC 在我们的初步结果中，野生型 Pg 细胞和纯化的 PGDHC，但不是丝氨酸棕榈酰。 转移酶 (SPT) KO Pg 细胞（PGDHC 生成不足），消除机械敏感性 Piezo1 介导 OC-genesis 的下调 数据表明 PGDHC 可能代表一种独特的毒力因子。 导致破骨细胞中 Piezo1 机械传感系统失调，从而导致骨发育迟缓 根据这些证据和初步结果，我们发现： 机械敏感性 Piezo1 在 OC 中引发细胞信号，诱导 IGF-1 的产生并抑制 OC- 起源，而 PGDHC 通过与 Myh9 结合，阻断了 Piezo1 的机械传感功能，因此， 减少受牙周炎影响的牙槽骨的骨再生潜力。 通过以下两个具体目标进行测试： 目标 1) 阐明 PGDHC- 的分子机制 体外 OC 上表达的 Piezo1 通道的抑制作用 目的 2) 确定 PGDHC 对体外表达的影响。 结扎和 Pg 诱导的牙周炎小鼠模型中 OC 表达的 Piezo1 四种主要训练。 候选人的目标包括： 1) 获得并提高与以下领域相关的先进科学方法的专业知识 骨免疫学和牙周病学，2) 提高复杂数据集的管理和分析技能，3) 获得新颖的图像采集和分析技术，将细菌毒力因子与 Piezo1 表达联系起来 使用拉曼光谱、体内成像、显微 CT 和生物层干涉测量法对破骨细胞进行研究，以及 4) 培养领导研究项目所需的技能，包括资助写作、实验室管理 人员、项目开发和执行。发起人 Kawai 博士和共同发起人 Han 博士。 在涉及牙周骨免疫学和免疫病理学的拟议研究中也具有丰富的经验 作为博士后研究员在学术研究领域职业发展的指导，将 致力于支持候选人的培训，并设有由5名专家组成的咨询委员会 脂质组学、骨生物学、机械感觉系统、微生物学和拉曼领域的研究人员 所有必需的设备、装置、协议和材料均可在 赞助商和顾问的实验室为候选人提供了一张桌子和实验室空间。 合作研究 (CCR) 是南佛罗里达州最大的生物医学研究大楼，配备 核心设施拥有多种尖端设备，可供考生使用 研究项目。