Nikoo Alizadeh
Pronouns: she/her
Research Mentor(s): Jason Knight
Co-Presenter:
Research Mentor School/College/Department: Internal Medicine/Rheumatology / Medicine
Presentation Date: April 20
Presentation Type: Oral5
Session: Session 5 – 3:40pm – 4:30 pm
Room: Breakout Room 3
Authors: Nikoo Alizadeh, Chao Liu
Presenter: 5
Abstract
Diabetes is a chronic disease that can be characterized by high blood glucose and it is one of the top leading causes of death in the United States. Recent evidence links its development to NETosis, the release of neutrophil extracellular traps (NETs) which is a vital part of the body’s immune system response to pathogens. Type 1 Diabetes Mellitus (T1DM) can be caused by a combination of insulin dysfunction or lack of secretion, and contributes to cardiovascular disease. Patients with T1DM experience hyperglycemia, which is related to an increase in NETosis as glucose uptake is essential for the process. Neutrophils have two kinds of extracellular glucose transporters: Glucose Transporters (GLUTs) and Sodium-Glucose Linked Transporters (SGLTs). Our aim in this study is to determine the most important glucose transporters in human neutrophils. By referencing a neutrophil RNA-seq database with the gene expression of various glucose transporters, we hypothesized that the most prominent transporters would be those with the highest gene expression. To collect data, we chose 11 of the top expressed genes to do RT-qPCR in order to validate the expression of the genes and conducted glucose uptake experiments to confirm the function of the transporters. To begin, neutrophils were isolated from healthy donor blood. The RNA is extracted from neutrophils and if it met the quality standards, it was converted into cDNA. Then, qPCR was conducted using SYBR green and the 2-??CT method. The GLUT-3 and SGLT-2 transporters had the highest mRNA expression at 9.5% and 3.9%respectively compared to ß-actin. Secondly, neutrophils were pretreated with the GLUT-1,2,3, 4 inhibitor, BAY-876, and the SGLT-2 inhibitor, Empagliflozin (EMPA). PMA was used to stimulate NETosis and glucose consumption, and 2-NBDG was used as a fluorescent tracer to monitor glucose uptake in the flow cytometry. The BAY-867 contributed to 64% inhibition of glucose uptake as well as 41% from EMPA. To summarize, GLUT-3 and SGLT-2 are possibly two of the most important glucose transporters.
Biomedical Sciences, Interdisciplinary
