Advisory Committee Chair
Advisory Committee Members
Mary Kathryn Sewell-Loftin
Date of Award
Degree Name by School
Master of Biomedical Engineering (MBE) School of Engineering
KLF2 is an important transcription factor for maintaining vascular homeostasis. Its expression in endothelial cells is triggered by shear stress. The endothelial glycocalyx also plays a role in regulating vascular function, and prior work has shown that glycocalyx expression is affected by shear stress as well. However, the direct link between KLF2 and mechanisms that regulate glycocalyx synthesis has not been established. In the present study, we used human lung microvascular endothelial cells (HLMVECs) to observe changes in glycocalyx expression, barrier function, and thromboinflammation caused by KLF2 overexpression. Lentiviral transduction was the method used for KLF2 overexpression, resulting in a 1.8-fold increase in KLF2 protein expression. Based on immunofluorescence imaging, the organization of the cytoskeleton was not affected by the level of KLF2 expression. Increased gene expression levels of heparan sulfate proteoglycans. Syndecan-3 and Syndecan-4 were observed in KLF2-transduced HLMVECs. Hyaluronan (HA) expression was decreased in KLF2 cells, which coincided with decreased gene expression of hyaluronan synthase-3, and increased gene expression of hyaluronidase. Based on a customized thromboinflammatory gene panel, KLF2 expressing cells showed lower expression of tissue factor and adhesion molecules, which may indicate a more anti-inflammatory environment. When the KLF2 expressing cells were treated with TNF-a, there was an increase in barrier function over 24 hours. ii However, relative to the untreated KLF2 cells, the resistance was decreased in the treated KLF2 cells, indicating less barrier function. Although our conclusions were limited by the relatively low level of KLF2 overexpression, some evidence of further research is warranted to clarify the role of KLF2 in glycocalyx expression, barrier function, and thromboinflammation.
O'Grady, Cambria, "KLF2 Regulation of Glycocalyx Expression, Barrier Function, and Thromboinflammation in Human Lung Microvascular Endothelial Cells" (2022). All ETDs from UAB. 132.