Abstract P310: Transcriptomic Profiling Of Sars-cov-2 Spike Protein S1 Subunit Mediated Changes In Human Vascular -pericytes, -endothelial Cells (ECs), -smooth Muscle Cells And Lymphatic ECs: Evidence For Differential Gene Regulation

Abstract

SARS-CoV-2 S1 spike protein (S1-SP) is increased in subjects with myocarditis after mRNA vaccine. However the underlying mechanism(s) of its vascular action are unclear. Using microarrays we assessed the effects of S1-SP on genes of vascular -ECs, -pericytes (PCs), -SMCs and LECs. Transcriptomic profiling of genes revealed that treatment of PCs, VECs, LECs and SMCs with 10ng/ml S1-SP for 36 hours upregulated 3935, 2512, 1422, 19 genes, respectively; and downregulated 3935, 3223, 1170, 37 genes, respectively. S1-SP significantly modulated genes in PCs, LECs and VECs, but not in SMCs. Pathway enrichment analysis highlighted modulation of TGF-β linked extracellular matrix, focal adhesion in PCS, VECs and LECs; whereas coronavirus disease, ribosome, cytoplasmic ribosomal protein, translation, protein metabolism, influenza infection, disease pathways were regulated in PCs and VECs, but not in LECs. A significant modulation in IL-2, cell senescence, IL4, TNFα and T-cell receptor associated apoptosis, Oncostatin M, p53 signaling was observed in LECs. Transcriptomic analysis showed induction of key COVID associated pro-inflammatory, coagulatory and matrix/adhesion molecules in PCs (PECAM1, CDH5, S1PR1, VWF, ESM1, EDN1, EFEMP1, ERG, ICAM2), but down regulation in VECs. Interestingly top genes that were down regulated in PCs (MYCT1, MMRN1, SEMA5A, GREM1, CEMIP, THBS2, BICC1, MYOCD, COL3A1, COL1A1, IGFBP3, PLPPR4, ITGA11, ADAMTS12, COL11A1, POSTN) were up regulated in VECs. Of all the genes regulated by S1-SP in PCs, VECs and LECs only two (FAP and P3H2) and eight (TMEM200A,MMP1, PLPP4, RGS5, TENM3, SULF1, COL12A1, SPOCK1) were up- and down regulated, respectively. DRG analysis highlighted contrasting modulation of genes associated with: myocarditis, metabolism (Glycolysis, one carbon metabolism, urea cycle and oxidative phosphorylation) and inflammation in PCs and VECs. No change in DRGs was observed in SMCs, whereas compared to PCs mixed regulation was observed in LECs. Also, Proteome array generated cytokine profile mimicked the impact of S1-SP on pro-inflammatory genes in PCs, VECs and LECs. Our findings provide first evidence that S1-SP differentially modulates genes and cytokines in PCs, LECs, VECs.