Reduced nephrin tyrosine phosphorylation impairs podocyte force transmission and accelerates detachment in disease

Podocytes are specialized kidney cells that form the slit diaphragm (SD), an intercellular filtration barrier against plasma protein loss. The SD is subject to significant mechanical strain which can be amplified in disease, leading to podocyte detachment. One key SD protein that might intercept mechanical strain and transmit adhesion signals is nephrin, although its influence on podocyte force transmission remains uncharacterized. Using immunoblotting and elastic resonator interference stress microscopy (ERISM), we demonstrate that nephrin clustering induces rapid podocyte force transmission and adhesion protein activation (paxillin, FAK, and p130Cas), which require nephrin tyrosine phosphorylation at its three YDxV motifs. Furthermore, using a model of diabetic nephropathy to amplify mechanical stress in vivo, we show that abolishing phosphorylation at YDxV tyrosines leads to exacerbated proteinuria, glomerular hypertrophy, and podocyte detachment. Altogether, this study links nephrin phosphorylation with force transmission, which is likely critical to maintaining podocyte adhesion during many renal pathologies.