Osteoimmunology of Retarded Bone Regeneration in Periodontitis

牙周炎骨再生迟缓的骨免疫学

• 批准号: 10451354
• 负责人: TOSHIHISA KAWAI
• 金额: $ 7.06万
• 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: 2021-09-01 至 2022-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10451354
• 关键词: Administrative Supplement; Affinity; African American; Award; Binding; Biological Assay; Biology; Blood Platelets; Bone Regeneration; CD100 antigen; Cell Culture Techniques; Cell surface; Data Analyses; Doctor of Philosophy; Escherichia coli; Funding; Grant; Health; Interferometry; Laboratories; Ligands; Lipids; Mediating; Methods; Monitor; Monoclonal Antibodies; Osteoclasts; Parents; Performance; Periodontitis; Platelet aggregation; Porphyromonas gingivalis; Psychology; Recombinants; Research; Research Project Grants; Role; Stains; Statistical Data Interpretation; Surface; TNFSF11 gene; Time; Training; Travel; Universities; Up-Regulation; Vimentin; college; dihydroceramide; experience; meetings; osteoclastogenesis; osteoimmunology; programs

This application is written to request for the one-year support for African American Post Bachelorette Trainee, Ms. Roodelyne Pierrelus under the program of “Research Supplements to Promote Diversity in Health-Related Research (PA-21-071).” This Research Administrative Supplements is requested to supplement the active RO1 grant project (PI, Kawai) entitled “parent RO1 grant, titled, “Osteoimmunology of regarded bone regeneration in periodontitis.” After she graduated from College of Psychology, Nova Southeastern University (NSU), on May 10, 2019, she started working in Kawai Laboratory on the study that evaluate the effects of P. gingivalis’ unique lipid, phosphoglycerol dihydroceramide (PGDHC), as well as E. coli LPS and Pg LPS on the platelet aggregation. She demonstrated outstanding performance in the above noted research project, and received IADR Bloc travel award twice for the abstract submitted to IADR/AADR meeting (March 2020, as well as July 2021). In the coming year, she will expand her research scope of platelet’s role in periodontitis to involve osteoclast biology. Parallelly, she will be working toward acceptance in graduate program of MD/PhD or equivalent. As such, we request funding to support her full-time research experience for one year. Background information: We discovered that activated platelets can promote the osteoclastogenesis in part by the expression of Semaphorin 4D (Sema4D) expressed on its surface. However, the neutralization of Sema4D with anti-Sema4D-mAb only partially inhibited the platelet-mediated upregulation of osteoclastogenesis. We learned that vimentin, the putative ligand for OC-STAMP is also expressed on the cell surface of activated platelets. Hypothesis: We hypothesized that vimentin expressed on activated platelets can act on OC-STAMP expressed by osteoclast precursors and promote their RANKL-induced osteoclastogenesis. Project Objectives for Candidate: Objective 1: To establish the role of Vimentin expressed on the activated platelets in the promotion of RANKL-induced osteoclastogenesis. Activated platelets will be co-cultured with RANKL- osteoclast precursors in the presence or absence of anti-OC-STAMP-mAb, anti-Vimentin-polyclonal Ab or control mAb/Ab. Osteoclastogenesis will be determined by q-PCR, TRAP staining and pit-formation assay. Using a biolayer interferometry (BLI), the binding affinity between OC-STAMP and activated platelet or recombinant vimentin will be monitored. We will also analyze whether platelet-derived vimentin is citrullinated. If so, the impact of Vimentin-citrullination on OC-STAMP-mediated osteoclastogenesis will be evaluated. After completing these two objectives, the candidate will have mastered the methods of cell culture, W- blotting, qPCR, BLI, and osteoclastogenesis assays, as well as basic statistical analysis and interpretation of data which strengthen her credential to be a candidate for graduate program of MD/PhD.

本申请是为了要求为非裔美国人邮政学员提供一年的支持， Roodelyne Pierrelus女士在“研究补充剂计划中促进与健康相关的多样性的计划 研究（PA-21-071）。”要求此研究管理补充剂来补充活跃的 RO1赠款项目（PI，Kawai）名为“父级RO1 Grant，标题为“骨骼的骨气不明” 牙周炎的再生。 （NSU），2019年5月10日，她开始在Kawai实验室进行研究，以评估P的影响。 牙龈牙龈的独特脂质磷酸甘油二氢可酰胺（PGDHC），以及大肠杆菌LPS和PG LPS 血小板聚集。她在上述研究项目中表现出了出色的表现， 接受IIADR/AADR会议的摘要两次获得IADR Bloc旅行奖（2020年3月） 如2021年7月）。在来年，她将扩大血小板在牙周炎中作用的研究范围 涉及破骨细胞生物学。可行的，她将努力接受MD/PhD研究生课程 或等效。因此，我们要求资金支持她一年的全日制研究经验。 背景信息：我们发现活化的血小板可以促进骨质碎屑的部分发生 通过在其表面上表达的Semaphorin 4D（SemA4D）的表达。但是，谈判 SEMA4D具有抗SEMA4D-MAB仅部分抑制了血小板介导的上调 破骨细胞生成。我们了解到Vimentin，OC-Stamp的假定配体也在细胞上表达 活化血小板的表面。 假设：我们假设在活化血小板上表达的波形蛋白可以作用于OC stamp 由破骨细胞前体表达，并促进其RANKL诱导的破骨细胞生成。 候选人的项目目标： 目标1：确定在激活血小板促进中表达的波形蛋白的作用 RANKL诱导的破骨细胞生成。活化的血小板将与ranklosteoplast共同培养 在存在或不存在抗OC-Stamp-mab的情况下，抗Vimentin-Polyclonal AB或对照 mab/ab。破骨细胞生成将由Q-PCR，陷阱染色和坑形化测定法确定。使用 Biolayer干扰（BLI），OC-Stamp与活化血小板或重组的结合亲和力 波形蛋白将受到监测。我们还将分析血小板衍生的波形蛋白是否为瓜氨酸。如果是这样， 将评估波形蛋白 - 硝化作用对OC-Stamp介导的破骨细胞生成的影响。 完成这两个目标后，候选人将掌握细胞培养方法，W- 印迹，QPCR，BLI和破骨细胞生成测定法，以及基本的统计分析和解释 增强她的证书的数据，成为MD/PHD研究生课程的候选人。