Podocyte-specific YAP activation triggers crescentic glomerulonephritis through cross-talk with parietal epithelial cells

YAP, a transcriptional co-activator, is known to control programs that are both cell- and context-dependent. We report that the overactivation of YAP (YAP5SA) in podocytes has both cell autonomous and non-cell autonomous consequences. RNA-seq analysis of YAP5SA-expressing compared to control podocytes revealed pathways and candidate genes that could explain both these effects. Overall design: To decipher the podocyte-specific transcriptional programs driven by YAP5SA, we generated human podocyte-derived cell lines stably expressing either pCW-(FLAG-NLS-)YAP5SA or pCW-eGFP (as a control), where transcription of the gene of interest is tetracycline inducible (Tet-On). This cell line, which was established by Saleem et al., is transformed by a thermolabile large-T antigen (ts SV40LT), so that these cells proliferate at 33 °C (active large-T antigen) but halt proliferation at 37 °C (inactive large-T antigen) and re-differentiate into podocytes over 10-14 days. To selectively overexpress (FLAG-NLS-)YAP5SA or GFP only in differentiated podocytes, cells were cultured at 37 °C for 12 days prior to tetracycline treatment. These differentiated podocytes were then treated with ± doxycyline (Dox) 0.1 µg for 24 hours.