Osteoimmunology of Retarded Bone Regeneration in Periodontitis

牙周炎骨再生迟缓的骨免疫学

• 批准号: 10451355
• 负责人: TOSHIHISA KAWAI
• 金额: $ 7.43万
• 依托单位: NOVA SOUTHEASTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-01 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10451355
• 关键词: Administrative Supplement; Attenuated; Basic Science; Biological Assay; Bone Regeneration; Bone Resorption; Cell Culture Techniques; Cells; Coculture Techniques; Data; Data Analyses; Dental; Dentistry; Flow Cytometry; Future; Gifts; Goals; Grant; Hispanics; Knowledge; Mediating; Methods; Monitor; Monoclonal Antibodies; Mouth Diseases; Osteoclasts; Parents; Pathogenicity; Periodontitis; Porphyromonas gingivalis; Proteins; Reproducibility; Research Project Grants; Role; Science; Scientist; Statistical Data Interpretation; TNFSF11 gene; Training; Translational Research; Vesicle; Vimentin; career development; osteoclastogenesis; osteoimmunology; precursor cell; programs; receptor; responsible research conduct

This application for Diversity Administrative Supplement (PA-21-071) is written to request for the one-year support for Hispanic Post Bachelorette Trainee, Ms. Elizabeth Leon, under the parent RO1 grant, titled, “Osteoimmunology of regarded bone regeneration in periodontitis.” She will perform the basic research to elucidate the mechanism underlying the pathogenic bone resorption in periodontitis. Through the proposed training, she will not only become familiar with the operational approach to perform basic research related to the oral diseases, but also master the conceptual approach to find the knowledge gap and the way to solve the existing problem. The fundamentals for increasing the rigor and reproducibility, as well as the principle for responsible research conduct will be also learned. Parallelly, she will be working toward acceptance in the program of dentistry. The goal of this training is to support the career development of a future clinician/scientist in the basic and translational research in the dental sciences. Candidate’s research project background: Our unpublished data demonstrated that P. gingivalis’ vesicles can promote the osteoclastogenesis which was significantly attenuated by the vesicles isolated from PAD-KO P. gingivalis. Furthermore, P. gingivalis’ vesicles up-regulated the expression of OC-STAMP in the RANKL- stimulated RAW264.7 osteoclast precursor cells. Hypothesis: We hypothesized that PAD expressed by vesicles of P. gingivalis can citrullinate the host derived Vimentin which, in turn, acts on OC-STAMP expressed by osteoclasts and promotes osteoclastogenesis Project Objectives for Candidate: Objective 1: To evaluate the role of PAD expressed by vesicles of P. gingivalis (Pg vesicle-PAD) in citrullinating the host proteins, including Vimentin, in the co-culture of vesicles and osteoclasts. Objective 2: To examine whether Pg vesicle-PAD-mediated citrullination of host vimentin can alter Vimentin’s effect on promoting RANKL-induced osteoclastogenesis. The vesicles will be isolated from PAD-KO Pg W83 (gift from Dr. Potempa) and WT Pg W83 using a ultracentrifuge machine. Those isolated vesicles, as well as whole bacterial cells, will be co-cultured with osteoclast precursors. The level of citrullination of host proteins will be determined by W-blot and Mass-Spec. The expression of PAD by vesicles will be monitored using a W-blot. Finally, possible engagement of OC- STAMP expressed by osteoclasts which is the receptor for citrullinated Vimentin in osteoclastogenesis will be determined by blocking with anti-OC-STAMP mAb or anti-citrullinated Vimentin mAb. After completing these two objectives, the candidate will have mastered the methods of cell culture, W- blotting, qPCR, flow cytometry, and osteoclastogenesis assays, as well as basic statistical analysis and interpretation of data which strengthen her credential to be a candidate for graduate program.

该多样性行政补充申请（PA-21-071）是为了要求一年的要求的 支持西班牙裔邮政学员的支持，伊丽莎白·莱昂女士，在父母的RO1 Grant下，标题为 “牙周炎中考虑的骨再生的骨免疫学。”她将进行基础研究 阐明牙周炎病原骨骼的基础机制。通过拟议的 培训，她不仅会熟悉执行与 口腔疾病，但也掌握了找到知识差距的概念方法和解决方案的方法 现有问题。增加严格和可重复性的基本原理，以及 负责任的研究行为也将学习。可行，她将努力接受 牙科计划。该培训的目的是支持未来临床/科学家的职业发展 在牙科科学的基本和翻译研究中。 候选人的研究项目背景：我们未发表的数据表明，牙龈疟原虫的蔬菜 可以促进破骨细胞生成，从PAD-KO中分离出的蔬菜显着减弱 牙龈疟原虫。此外，牙龈疟原虫的蔬菜上调了OC stamp在rankl-的表达 刺激的RAW264.7破骨细胞前体细胞。 假设：我们假设由牙龈疟原虫蔬菜表达的垫可以citrullinate citrullinate衍生的宿主 波形蛋白反过来作用于由破骨细胞表达并促进破骨细胞发生的OC stamp蛋白 候选人的项目目标： 目的1：评估用牙龈假单胞菌蔬菜（pg囊泡）表达的垫的作用 在蔬菜和破骨细胞的共培养中，在包括波形蛋白在内的宿主蛋白，包括波形蛋白。 目标2：检查宿主波形蛋白的pg囊泡 - 帕 - 帕德介导的柠檬酸是否可以改变 波形蛋白对促进RANKL诱导的破骨细胞生成的影响。 蔬菜将与PAD-KO PG W83（Potempa博士的礼物）和WT PG W83隔离 超速离心机。那些孤立的蔬菜以及整个细菌细胞将与 破骨细胞前体。宿主蛋白的柠檬化水平将由W blot和质量规格确定。 蔬菜的表达将使用w blot监测。最后，可能参与OC- 破骨细胞表达的邮票是破肉地核酸藻蛋白的接收器，将是破骨细胞发生的 通过用抗OC-Stamp MAB或抗硝化的波形蛋白mAb封闭确定。 完成这两个目标后，候选人将掌握细胞培养方法，W- 印迹，QPCR，流式细胞仪和破骨构成作用测定法，以及基本的统计分析和 对数据的解释可以增强她的证书成为研究生课程的候选人。