Molecular Profiling of Fetal and Adult Liver Sinusoidal Endothelial Cells: A F8 Secreting Cell

In human, F8, whose deficiency leads to hemophilia A, is largely synthesized and secreted by the sinusoidal endothelial cells of the liver. However, the characteristics of the cells suitable for this process are not well known.

In this study, we performed genome-wide expression (microarrays) and CpG methylation profiling (microarrays and whole genome bisulfite sequencing) of fetal and adult human liver sinusoidal endothelial cells (LSECs) together with other fetal endothelial cells, from lung (microvascular and arterial), heart (microvascular), and umbilical origin. Our results reveal specific expression and methylation profile distinguishing LSECs at both fetal and adult stages. Differential gene expression of fetal LSECs in comparison to other fetal ECs pointed to differential regulated pathways and biofunctions in fetal LSECs were EIF2; signaling is downregulated, while antigen presentation pathway and cellular movement are activated.

We used targeted bisulfite resequencing to confirm selected differentially methylated regions, between fetal LSECs and other fetal ECs, and found that loci at GSTO2 and ZC3H12D provided suitable transition epigenetic markers that show gradual increase and decrease in methylation, respectively, from fetal non-LSECs to fetal LSECs and to adult LSECs. We used the selected methylation markers to test the degree of similarity of in house in vitro generated vascular endothelial cells to different fetal or adult LSECs; a higher similarity was found to the fetal than to adult LSECs. Our molecular profiling study provided the base of future studies that lead to (1) the in vitro derivation of LSECs (by understanding the gene regulatory networks), (2) boost the secretion of F8 (by studying the expression of TFs binding to F8 promoter), and (3) provide a guide to test the effectiveness of in vitro differentiated LSECs (by comparing expression and methylation markers) that will be used in cellular therapies.

Publication History

Article published online:
13 November 2020