Gene expression profiles of human podocyte cell line treated with and without ANGPT2

Glomerular diseases are the leading cause of chronic kidney diseases with pathomechanisms largely unclear. It is known that ANGPT2 regulates endothelial cell homeostasis and function via TEK/TIE2 and its deregulation causes endothelial damage. We found that ANGPT2 is upregulated in glomerular diseases and wondered whether it has any effect on glomerular podocytes and mesangial cells given that they have no or low TEK expression. We treated podocytes and mesangial cells in culture with ANGPT2 and found no overt cellular changes. RNA-seq analysis showed that gene expression was altered in both podocytes and mesangial cells and that the regulated genes in the two cell types were fundamentally different. GO and KEGG analyses showed that the two groups of regulated genes were enriched in distinct processes and pathways. These results suggest that ANGPT2 exerts effects on both podocytes and mesangial cells and that increased ANGPT2 may be involved in glomerular injury by affecting podocytes and mesangial cells in addition to endothelial cells. Immortalized human podocytes from Dr. Saleem M’s laboratory (University of Bristol, Bristol, UK) were cultured with RPMI1640 medium supplemented with 10% FBS, 1% Pen/Strep and 1% ITS at 33°C in 5% CO2 incubator for growth, followed by trypsinization, replating and culture at 37°C for 10 days to obtain differentiated cells. Then the differentiated podocytes were either treated with 500 ng/ml recombinant ANGPT2 (MCE, HY-P7510) or left untreated with only vehicle for 24 hours. The cells were homogenized with Trizol (Invitrogen) and the total RNA was extracted following the manual instruction. The RNA was then subject to RNA-seq.