ABSTRACT: SARS-CoV-2 S1 spike protein (S1-SP) plays a critical role by actively facilitating host and virus interaction via ACE2 to mediate the deleterious effects of COVID19. Evidence suggests that SP devoid of virus is not latent and modulate proinflammatory actions. Circulating SP levels increase in subjects recieving mRNA vaccines and those who develop post-vaccination myocarditis. However the mechanism(s) involved remain unclear. Since interaction between circulating factors and vascular cells play a key role in the pathophysiology of cardiovascular complication, using microarrays we assessed the modulatory effects of S1-SP on genes of key vascular cells (human vascular and lymphatic endothelial cells (VECs; LECs), -pericytes (PCs), and –smooth muscle cells (SMCs). Transcriptomic profiling of genes from microarray studies 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 of modulated genes showed significant regulation of several pathways, including TGF-β regulation of extracellular matrix, focal adhesion, pathways in cancer 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. Moreover, interleukin-2 and cellular senescence pathways were significantly regulated in both LECs and VECs. In LECs, significant modulation in IL-4 regulation of apoptosis, Oncostatin M, T cell receptor regulation of apoptosis, TNF-α effects on cytokine activity/cell motility & apoptosis, p53 signaling and fluid shear stress and atherosclerosis was also observed. 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 downregulation in VECs. Interestingly top genes that were downregulated in PCs (MYCT1, MMRN1, SEMA5A, GREM1, CEMIP, THBS2, BICC1, MYOCD, COL3A1, COL1A1, IGFBP3, PLPPR4, ITGA11, ADAMTS12, COL11A1, POSTN) were upregulated in VECs. Of the top 20 genes up or down -regulated in LECs 7 were similar to PCs and 4 to VECs, suggesting that S1-SP induces differential actions in vascular cells. 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 crbon metabolism, urea cycle and oxidative phosphorylation) and inflammation in PCs and VECs. No modulation of DRGs was observd in SMCs whereas mixed regulation compared to PCs was observd in LECs. Taken together our findings provide first evidence that S1-SP differentially modulates genes in PCs, LECs, VECs. Importantly it modulates key genes and associated with the pathophysiology of myocarditis, mitochondrial metabolism, inflammation and Covid associated genes. The comparative transcriptomic profiling of S1-SP induced changes in vascular PCs, VECs, LECs and SMCs, provides important insights to elucidate the mechansims mediating the deleterious CV actions of circulating S1-SP associated with COVID-19 and /or mRNA vaccine.
