ARHGEF7 (-PIX) Is Required for the Maintenance of Podocyte Architecture and Glomerular Function.

BACKGROUND

Previous studies showed that Cdc42, a member of the prototypical Rho family of small GTPases and a regulator of the actin cytoskeleton, is critical for the normal development and health of podocytes. However, upstream regulatory mechanisms for Cdc42 activity in podocytes are largely unknown.

METHODS

We used a proximity-based ligation assay, BioID, to identify guanine nucleotide exchange factors that activate Cdc42 in immortalized human podocytes. We generated podocyte-specific ARHGEF7 (commonly known as -PIX) knockout mice by crossing -PIX floxed mice with Podocin-Cre mice. Using shRNA, we established cultured mouse podocytes with -PIX knockdown and their controls.

RESULTS

We identified -PIX as a predominant guanine nucleotide exchange factor that interacts with Cdc42 in human podocytes. Podocyte-specific -PIX knockout mice developed progressive proteinuria and kidney failure with global or segmental glomerulosclerosis in adulthood. Glomerular podocyte density gradually decreased in podocyte-specific -PIX knockout mice, indicating podocyte loss. Compared with controls, glomeruli from podocyte-specific -PIX knockout mice and cultured mouse podocytes with -PIX knockdown exhibited significant reduction in Cdc42 activity. Loss of -PIX promoted podocyte apoptosis, which was mediated by the reduced activity of the prosurvival transcriptional regulator Yes-associated protein.

CONCLUSIONS

These findings indicate that -PIX is required for the maintenance of podocyte architecture and glomerular function Cdc42 and its downstream Yes-associated protein activities. This appears to be the first evidence that a Rho-guanine nucleotide exchange factor plays a critical role in podocytes.