Abstract 1265 iPSC-derived endothelial cells from ACS patients and healthy controls show significant differences in their inflammatory response assessed via RNA-sequencing and basal DNA-methylation patterns

Clin Res Cardiol (2023). https://doi.org/10.1007/s00392-023-02180-w
iPSC-derived endothelial cells from ACS patients and healthy controls show significant differences in their inflammatory response assessed via RNA-sequencing and basal DNA-methylation patterns
E. T. Strässler¹, N. Kränkel¹, U. Landmesser¹
¹CC 11: Med. Klinik für Kardiologie, Charité - Universitätsmedizin Berlin, Berlin;
BACKGROUND: Cardiovascular diseases (CVD) remain the leading cause of death worldwide and, as multifactorial disease entities, their development and progression are strongly impacted by goth genetic predisposition and lifestyle. Induced pluripotent stem cells (iPSC) are an ideal tool to differentiate (epi-) genetics from lifestyle-related disease risk modulation and identify the underlying molecular mechanisms. AIM: The study aims to determine differences in iPSC-derived endothelial cell characteristics and functionality between healthy donors and high-risk CAD patients with a suspected genetic predisposition to disease development and progression. METHODS: iPSC from participants with different disease statuses (healthy > 65 y/o, acute coronary syndrome (ACS) < 65y/o) were differentiated into iPSC-EC. Cells were exposed to inflammatory stimulation via tumour necrosis factor-alpha (TNF-a) followed by bulk RNA-seq. Monocyte (THP-1) recruitment was assessed under beneficial and detrimental flow conditions. Mitotic age was calculated based on DNA-methylation via Hannum estimate for each donor at the iPSC and iPSC-EC stages. RESULTS: ACS-iPSC-EC had significantly stronger upregulation of E-selectin upon TNF-a stimulation on mRNA level (ACS: 532.1-fold±48.7 vs healthy: 322.3-fold±55.7 with p=0.02). ICAM1 protein upregulation was stronger in ACS-iPSC-EC as determined by flow cytometry (ACS: 1.4-fold±0.01 vs healthy: 1.1-fold±0.0007; p<0.001). ACS-iPSC-EC recruited significantly more THP-1 per view field under atheroprotective (high shear stress, laminar) flow conditions (ACS: 78.4±20.7 vs healthy: 33.25±12.3 with p=0.04) compared to healthy iPSC-EC. RNA-seq showed significant differences in TNF-a-induced transcriptome changes. Gene set enrichment analysis identified differences in pathways related to leukocyte and lymphocyte-mediated immunity, adaptive immune response, and cytokine activity, among others. DNA methylation analysis showed differences in basal DNA methylation. IPSC of both groups had a negative calculated mitotic age (in years) which significantly increased after endothelial differentiation and four population doublings (ACS-iPSC: -14.1±0.7 vs ACS-EC: -0.1±1.1, p<0.0001 and healthy-iPSC: -13.7±0.8 vs healthy-EC: 2.5±1.5, p=0.0001). CONCLUSION: The higher rate of basal THP-1 recruitment, the higher upregulation of adhesion molecules on mRNA and protein level, as well as the distinct transcriptome and DNA methylation changes, suggest intrinsic differences, eg in inflammatory pathways, at rest and upon activation and show that iPSCs may also be used as in vitro models for polygenetic diseases such as CAD.
https://dgk.org/kongress_programme/jt2023/aP1705.html