Höfundar:
Arnar Ingi Vilhjálmsson, Óttar Rolfsson, Adrián López García de Lomana, Eva Jacobsen, Sarah McGarrity
Patients suffering from shock-induced endotheliopathy (SHINE) share phenotypic features of endothelial dysfunction and sustained high levels of circulating catecholamines. These phenotypic features correlate with poor disease outcome and are statistically associated with development of organ failure and death. The molecular response mechanism of endothelial cells contributing to the disease entity is poorly understood. Alterations in the transcriptome are observed upon high levels of catecholamines. Four gene targets involved in cell proliferation show a significant increase in transcription: GRAMD1B, DIO2, AREG and NR4A3. The role of these genes in relation to SHINE is yet to be clarified.
Endothelial cells treated temporally with catecholamines represent an in vitro cell model to study SHINE. We are using CRISPR-Cas9 knock-out cell lines to investigate the function of selected genes in SHINE. Furthermore, seahorse assays and liquid chromatography-mass spectrometry (LC-MS) are being used to examine the changes in energy demand and map metabolic changes of endothelial cells following catecholamine stimulation.
Endothelial cells treated with catecholamines change the energy demand of the cells, driving up oxygen consumption and glycolysis. Alterations in the proteome are also observed, further supporting the transcriptomic data. Knockout cell lines have now been successfully generated for selected genes to study their role in the stress response that accompanies SHINE.
The transcriptomic and metabolomic response in endothelial cells to catecholamine stimulation has been defined and further functional studies of selected genes important to this process are pending.
