Research
|
Working as an undergraduate researcher in the Weinstein Lab was a challenging and rewarding experience. This
research project allowed me to engage with classroom concepts in a hands-on and
memorable way by applying my biology, genetics, immunology, and neurobiology to
better understand the phenomenon of ischemic preconditioning. Doing my own
project expanded my capacity for critical thinking and creative problem-solving,
as I was encouraged to take initiative for my experiments and think
independently while working within the framework provided by my mentors, Dr.
Jonathan Weinstein (MD/PhD) and research scientist Richard Lee. They taught me
technical skills, and we met weekly to review my proposed experimental setups
and discuss my results and how my project complemented the work of other lab
members. This experiential learning was valuable for gaining technical skills,
as well as a sense of the research process and the team dynamics of lab work.
Seeing how Dr. Weinstein balances clinical medicine with research to improve
patient outcomes made me excited about my future, as I hope to incorporate
research into my career as a doctor. Below is my abstract and poster for the UW Undergraduate Research Symposium.
Abstract:
Ischemic preconditioning (IPC) is the neuroprotective phenomenon in which a brief ischemic episode induces genetic reprogramming of affected brain cells, causing physiological changes that minimize cell death from subsequent, sustained ischemic insult. We hope to characterize the cellular and molecular mechanisms of IPC. Our lab previously has shown that IPC is dependent on Toll-Like Receptor 4 (TLR4) and involves the production of interferon stimulated genes (ISGs) in microglia. Following ischemia, damaged neurons release endogenous TLR4 agonists called damage-associated molecular patterns (DAMPS), which induce interferon (IFN) release by microglia. The IFNs initiate a biochemical signaling cascade that activates transcription factors which bind to the interferon-stimulated response element (ISRE), promoting ISG transcription which alters the immunological function and phenotype of microglia, mediating neuroprotection. My project is to confirm the cell signaling mechanism by which hypoxic/hypoglycemic (H/H) conditions induce ISG expression in vitro. To accomplish this, I am using a reporter assay to detect ISRE activation with luciferase protein expression, looking specifically at the effects of H/H experimental stimulations on ISRE activity in microglia from the N9 cell line. I am first optimizing a transfection protocol using green fluorescent protein (GFP) as a reporter. I will use the optimized protocol for experiments, transfecting microglia with DNA encoding the luciferase enzyme linked to the ISRE promoter, then incubating the cells under H/H conditions, and finally using a plate reader to quantify fluorescence. We anticipate that H/H stimulation will result in increased luminescence in wild-type microglia, and we would interpret these results as indicating that ISG expression in H/H-stimulated microglia is mechanistically due to transcriptional activation at ISRE sites. Identifying the genetic actors in IPC will reveal intracellular targets for pharmaceuticals that can mimic this neuroprotection. We can then administer these medications pre-emptively to patients at risk for stroke or emergently to acute stroke patients. |
Visit other pages on this pathway: