As a student with strong interest in pursuing a career in biochemistry, Sylvia has set her sights on earning a PhD in biochemistry. Through further research and laboratory experience, she hopes to make a positive impact towards human health and disease research. Sylvia’s unwavering commitment to scientific inquiry and their determination to grow and learn new fields makes her a promising scholar. In addition to her academic pursuits, Sylvia enjoys spending her free time outside exploring the Spokane area in the summer with her family and taking road trips to National parks. Her favorite travel destination was the cloud rainforests of Costa Rica.
2023 EWU Faculty Research Mentor: Dr. Jason Ashley
Research Title: Overexpression of Fringe syltransferase in osteoclastogenesis
Abstract: Osteoclasts, the multinucleated cells crucial for bone resorption, are a critical component in both physiological and pathological skeletal remodeling. Osteoclastogenesis, the process of differentiation of the monocyte/macrophage originating precursors to osteoclasts, relies heavily on RANK signaling. However, the size, activity, and lifespan can be influenced by other pathways,such as Notch signaling. Notably, Notch signaling is regulated by selective glycosylation through the Fringe family of O-fucose-specific ß1,3-N-acetylglucosaminyltransferases (O ß1,3 GnTs). To investigate the impact of individual Fringe glycosyltransferases on osteoclastogenesis, we conducted experiments exploring the overexpression of the individual Fringe glycosyltransferases [Lunatic (LFNG), Manic (MFNG), and Radical (RFNG)] found to influence mammal Notch pathways. These full-length coding sequences were obtained from a mouse bone marrow cDNA library and inserted into the pMXs-Puro vector, the Moloney Murine Leukemia Virus (MMLV)-derived gammaretroviral system. Retroviral particles were generated through transient co-transfection with VSV-G envelope plasmids into Platinum-E packaging cells. Following infection of RAW264.7 cells, immortalized macrophage/osteoclast precursors, and primary cells collected from male C57BL/6 mice we confirmed overexpression via RT-PCR, and subjected variable seeding methods to RANKL and MCSF in primaries to stimulate osteoclastogenesis. At the conclusions of differentiation, we stained cells with acridine orange (AO) stain as well as for tartrate-Resistant Acid Phosphatase (TRAP) activity to assess resulting morphological variations. We found that LFNG-overexpressing (OE) cell lines in primary cultures did[...]
