Wcn26-289 a Vesicular Trafficking Pathway Mediated by N-Ethylmaleimide-Sensitive Factor Governs Integrin Maturation and Podocyte Integrity

Introduction:The adhesion of podocytes-terminally differentiated epithelial cells-to the glomerular basement membrane is critical for kidney function.While adhesion disruption causes podocyte loss, the molecular mechanisms by which impaired focal adhesion drives progressive podocyte dysfunction and cell loss in glomerulosclerosis remain poorly understood.Methods: We used doxycycline-inducible podocyte-specific Tln1 knockout (Tln1-cKO) mice, an established model of impaired focal adhesion, and performed single-cell RNA sequencing to identify early molecular changes.To investigate a key candidate gene, we generated podocyte-specific Nsf knockout (Nsf-cKO) mice and assessed them using cell spreading assays, morphological and molecular analyses, and live-cell imaging using the Retention Using Selective Hooks (RUSH) system.Results: Screening of Tln1-cKO podocytes revealed Nsf, which encodes N-ethylmaleimide-sensitive factor (NSF)-an AAA+ ATPase essential for vesicular trafficking by disassembling soluble NSF attachment protein receptor complexes-as one of the most significantly downregulated genes in injured podocytes.Podocyte-specific Nsf deletion resulted in massive proteinuria, podocyte loss, glomerulosclerosis, tubulointerstitial fibrosis, renal failure, and early mortality by 3 weeks.Nsf-cKO podocytes exhibited impaired cell spreading, with defective