Single‐Cell RNA Sequencing‐Based Identification of Novel Podocyte Injury‐Associated Genes From Isolated Glomeruli of Diabetic db/db Mice

ABSTRACT Podocyte injury is a primary driver of progression in diabetic kidney disease (DKD). Although single‐cell RNA sequencing (scRNA‐seq) is a powerful tool for identifying novel disease‐associated genes, podocytes are frequently underrepresented in whole‐kidney scRNA‐seq datasets because they constitute only a minor fraction of renal cells. To overcome this limitation, we performed scRNA‐seq on magnetically isolated glomeruli from diabetic db/db mice, generating a high‐resolution, glomerulus‐focused transcriptomic dataset. Diabetic glomeruli exhibited a marked reduction in podocyte abundance and extensive transcriptional remodeling. By integrating in vivo scRNA‐seq data with in vitro analyses of cultured human podocytes exposed to diabetic stimuli—specifically palmitate‐induced lipotoxicity—we identified four candidate genes ( OPCML , ADM , NELL2 , and DHRS3 ) associated with diabetic podocyte injury. Among these candidates, in vitro mechanistic validation demonstrated that knockdown of OPCML or ADM , as well as overexpression of DHRS3 , significantly attenuated palmitate‐induced apoptosis and preserved actin cytoskeletal integrity. These findings identify OPCML , ADM , and DHRS3 as novel mediators of podocyte injury and promising therapeutic targets for DKD.