ARHGEF7 (<i>β</i>-PIX) Is Required for the Maintenance of Podocyte Architecture and Glomerular Function.

Abstract

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. 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 <i>β</i>-PIX) knockout mice by crossing <i>β</i>-PIX floxed mice with Podocin-Cre mice. Using shRNA, we established cultured mouse podocytes with <i>β</i>-PIX knockdown and their controls. We identified <i>β</i>-PIX as a predominant guanine nucleotide exchange factor that interacts with Cdc42 in human podocytes. Podocyte-specific <i>β</i>-PIX knockout mice developed progressive proteinuria and kidney failure with global or segmental glomerulosclerosis in adulthood. Glomerular podocyte density gradually decreased in podocyte-specific <i>β</i>-PIX knockout mice, indicating podocyte loss. Compared with controls, glomeruli from podocyte-specific <i>β</i>-PIX knockout mice and cultured mouse podocytes with <i>β</i>-PIX knockdown exhibited significant reduction in Cdc42 activity. Loss of <i>β</i>-PIX promoted podocyte apoptosis, which was mediated by the reduced activity of the prosurvival transcriptional regulator Yes-associated protein. These findings indicate that <i>β</i>-PIX is required for the maintenance of podocyte architecture and glomerular function <i>via</i> 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.