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，特别是牙龈卟啉单胞菌(Pg) 参与破坏牙周炎的牙齿支持组织。在产生的几个致病因子中 通过PG，一种独特的致病脂质，磷酸甘油二氢神经酰胺(PGDHC)，促进RANKL诱导的OC- Genesis通过作用于非肌肉肌球蛋白II-A(Myh9)，同时抑制OB生成因子IGF-1的产生 1，来自OCS。在我们的初步结果中，野生型PG细胞和纯化的PGDHC，但不是丝氨酸棕榈酸酯 Piezo1介导的机械敏感的转移酶(SPT)KO PG细胞(PGDHC合成缺陷) 对卵巢癌发生的下调。这些数据表明，PGDHC可能代表着一种独特的毒力因子， 破骨细胞中Piezo1-机械连接系统的失调导致骨发育迟缓 再生。根据这些证据和初步结果，我们假设 机械敏感的Piezo1在OCS中诱导细胞信号，诱导IGF-1的产生并抑制OC-1 Genesis，而PGDHC通过与Myh9结合，阻止Piezo1的机械传感功能，因此， 牙周炎影响牙槽骨的骨再生能力减弱。这一假设将是 目的1)阐明PGDHC的分子机制。 体外介导抑制OCS上表达的Piezo1通道。目的2)研究前列地尔对大鼠心脏功能的影响。 Piezo1在结扎和PG诱导的小鼠牙周炎模型中的表达四大培训 应聘者的目标包括：1)获得和改进与以下相关的先进科学方法方面的专业知识 骨免疫学和牙周学，2)提高管理和分析复杂数据集的技能，3) 获取新的图像采集和分析技术以连接细菌毒力因子和表达的Piezo1 在破骨细胞上，使用拉曼光谱、活体成像、显微CT和生物层干涉测量法，以及4) 培养领导研究项目所需的技能，包括拨款、实验室管理 人员、项目开发和执行。赞助商河井博士和共同赞助商韩博士，他们是 对拟议的牙周骨免疫学和免疫病理学研究有丰富经验 作为博士后研究员在学术研究领域职业发展的导师，将 致力于支持候选人的培训，以及由5名专家组成的咨询委员会 脂类组学、骨生物学、机械传感系统、微生物学和拉曼等领域的研究人员 光谱学。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。应聘者在中心有一张办公桌和一间实验室 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 在核心设施中拥有各种尖端设备，所有这些都可用于候选人的 研究项目。