Katherine Nguyen
BME PhD Proposal Presentation

Date:2022-06-27
Time: 12:00 PM-2:00 PM
Location / Meeting Link: EBB 4029 or https://gatech.zoom.us/j/99810303612?pwd=alBTS1V6Y2lScHNBM0tKSCtYSkIydz09

Committee Members:
Shuichi Takayama, PhD (Advisor) Jason Knight, MD, PhD Wilbur Lam, MD, PhD Rebecca Levit, MD Aniruddh Sarkar, PhD


Title: A High-throughput Degradation Assay Using Mimetic Neutrophil Extracellular Traps (NETs) to Study How DNase and Proteases Work Simultaneously to Clear NETs

Abstract:
Neutrophil extracellular traps (NETs) are decondensed chromatin with granule proteins released by neutrophils. While NETs were initially thought to be a bacteria killing method, interest in NETs grew as further research that NETs are involved in a large variety of pathologies such as sepsis, infections, autoimmune disorders, cardiovascular diseases, pulmonary diseases, and even cancer. In these conditions, over production or improper clearance of NETs were often tied to disease severity. Thus, there is a possibility for new treatments that target and degrade NETs. However more knowledge about NET degradation modulators is necessary to develop the treatments. Current degradation studies mostly focus on DNases found in serum despite a growing opinion of the possibility of protease mediated degradation. Therefore, the objective of this proposal is to create a high-throughput assay using mimetic NETs to study how serum DNases and proteases work simultaneously to clear NETs. In aim 1, we will create synthetic NET-like structures referred to as surface-webs to create a fluorescence-based assay suitable for high throughput studies. Then, we will characterize the role of common serum proteases on NET degradation. Finally, the surface-webs will be used to analyze NET degradation by proteases in clinical serum samples. Success of this proposal will identify a new target for NET degrading therapeutics and simultaneously create a convenient assay for further NET research.