254 - iPSC-derived endothelial cells from high-risk CAD patients demonstrate increased inducible adhesion molecule upregulation by inflammatory stimulation and gap closure delay compared to healthy controls (ID 1152)
Abstract
Background and Aims
Atherosclerosis remains a leading cause of death worldwide. Thus, it is imperative to not only illuminate the general pathomechanism but also investigate the underlying mechanisms in genetically predisposed patients. Here, induced pluripotent stem cells (iPSC) are an ideal tool to experimentally separate the impact of a donor´s genetic background from their lifestyle on cardiovascular disease risk in pathomechanistic studies.
Methods
IPSC were generated from participants with similar lifestyle but different disease progression (healthy > 65 y/o, stable CAD > 65 y/o, ACS < 65y/o) and differentiated into endothelial cells (iPS-EC). After quality control, IPS-EC were assessed after exposure to inflammatory stimulation via TNF-α under static and pro- vs. anti-atherogenic flow conditions (laminar vs. turbulent flow, high vs. low shear stress) regarding their gene and protein expression of adhesion molecules. Angiogenesis-related network-formation was assessed under static conditions.
Results
IPS-ECs were able to form network structures on extracellular matrix, albeit less efficiently than primary ECs (p<0.001). ACS-iPS-ECs showed a significantly stronger upregulation of E-selectin in response to inflammatory stimulation with TNF-a than healthy-iPS-ECs on mRNA level (ACS: 532.1-fold±48.7 vs. healthy: 322.3-fold±55.7, p=0.02). Similarly, ICAM1 protein upregulation was stronger in ACS-iPS-ECs than in healthy iPSC-ECs as assessed by flow cytometry (ACS: 1.4-fold±0.01 vs. healthy: 1.1-fold±0.0007; p<0.001) whereas VCAM1 upregulation was weaker in CAD/ACS-iPS-ECs compared to healthy controls (CAD/ACS: 1.8-fold±0.3 vs. healthy: 4.2-fold±0.9, p<0.001).
Conclusions
Upregulation of certain adhesion molecules (E-Selectin, ICAM-1) in response to inflammatory stimulation was more pronounced in iPS-ECs from ACS patients, supporting a stronger recruitment of pro-inflammatory leukocytes.
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