Modeling the Effects of Severe Metabolic Disease by Genome Editing of hPSC-Derived Endothelial Cells Reveals an Inflammatory Phenotype.
Filip RoudnickyYanjun LanMax FriesenGregor DernickJitao David ZhangAndreas StaempfliNatalie BordagAntje Wagner-GolbsKlaus ChristensenMartin EbelingMartin GrafMark BurcinClaas Aiko MeyerChad A CowanChristoph PatschPublished in: International journal of molecular sciences (2019)
The kinase AKT2 (PKB) is an important mediator of insulin signaling, for which loss-of-function knockout (KO) mutants lead to early onset diabetes mellitus, and dominant active mutations lead to early development of obesity and endothelial cell (EC) dysfunction. To model EC dysfunction, we used edited human pluripotent stem cells (hPSCs) that carried either a homozygous deletion of AKT2 (AKT2 KO) or a dominant active mutation (AKT2 E17K), which, along with the parental wild type (WT), were differentiated into ECs. Profiling of EC lines indicated an increase in proinflammatory and a reduction in anti-inflammatory fatty acids, an increase in inflammatory chemokines in cell supernatants, increased expression of proinflammatory genes, and increased binding to the EC monolayer in a functional leukocyte adhesion assay for both AKT2 KO and AKT2 E17K. Collectively, these findings suggest that vascular endothelial inflammation that results from dysregulated insulin signaling (homeostasis) may contribute to coronary artery disease, and that either downregulation or upregulation of the insulin pathway may lead to inflammation of endothelial cells. This suggests that the standard of care for patients must be expanded from control of metabolic parameters to include control of inflammation, such that endothelial dysfunction and cardiovascular disorders can ultimately be prevented.
Keyphrases
- endothelial cells
- signaling pathway
- cell proliferation
- oxidative stress
- early onset
- type diabetes
- crispr cas
- genome editing
- wild type
- coronary artery disease
- high glucose
- pluripotent stem cells
- glycemic control
- poor prognosis
- fatty acid
- single cell
- insulin resistance
- anti inflammatory
- end stage renal disease
- late onset
- vascular endothelial growth factor
- newly diagnosed
- metabolic syndrome
- stem cells
- ejection fraction
- healthcare
- bone marrow
- gene expression
- weight loss
- chronic kidney disease
- long non coding rna
- cardiovascular events
- staphylococcus aureus
- adipose tissue
- peripheral blood
- peritoneal dialysis
- candida albicans
- pseudomonas aeruginosa
- atrial fibrillation
- body mass index
- quality improvement
- left ventricular