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.

项目摘要/摘要 宿主对牙周病原体的免疫炎症反应，尤其是牙龈卟啉单胞菌（PG）是 从事牙周炎中牙齿支持组织的破坏。在产生的几种有毒因素中 由独特的致病性脂质PG，磷酸甘油二氢可胺（PGDHC）促进了RANKL诱导的OC- 通过作用于非肌肉肌球蛋白II-A（MYH9），同时抑制了ob生成因子的产生，IGF- 1，来自OCS。在我们的初步结果中，野生型PG细胞和纯化的PGDHC，但不含丝氨酸棕榈酰 转移酶（SPT）KO PG细胞（PGDHC生产缺乏），促进的机械敏感性压电介导 OC生成的下调。数据表明PGDHC可能代表一个独特的病毒因素 导致破骨细胞中压电1-元素系统的失调，导致骨骼智障 再生。基于这些证据和初步结果，可以假设 机械敏感的压电1引起OC中的细胞信号，该信号诱导IGF-1的产生并抑制OC- 创世纪，而PGDHC通过与MYH9的结合来阻止Piezo1的机理功能，因此， 减少受牙周炎影响的牙槽骨的再生潜力。这个假设将是 通过以下两个特定目的测试：目标1）阐明PGDHC-的分子机制 介导的对OCS在体外表达的压电通道的抑制作用。目标2）确定PGDHC对 OC在绑扎和PG诱导的牙周炎的小鼠模型中表达的压电1。主要的四个训练 候选人的目标包括：1）获得和提高与高级科学方法相关的专业知识 骨气免疫学和牙周病学，2）提高复杂数据集管理和分析的技能，3） 获取新的图像采集和分析技术，以将细菌病毒因子和表达的Piezo1连接起来 在破骨细胞上，使用拉曼光谱，体内成像，微CT和Biolayer干涉法，4） 开发领导研究计划所需的技能，包括赠款写作，实验室管理 人员，项目开发和执行。赞助商Kawai博士和共同赞助商Han博士 在拟议的研究中经验丰富的牙周骨气免疫学和免疫病理学 作为博士后研究员在学术研究领域的职业发展的心理，将是 致力于支持候选人的培训，以及由5个专家组成的咨询委员会 脂质组学，骨生物学，机械学系统，微生物学和拉曼领域的研究人员 光谱法分别。所有必需的设备，设备，协议和材料都可以在 赞助商和顾问实验室。为候选人提供了一个桌子和实验室空间 合作研究（CCR）是南佛罗里达州最大的生物医学研究大楼 核心设施中有各种尖端的设备，所有这些设备均可用于候选人的 研究项目。