Research Project
10451354
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.
