LncRNA evf‐2 Exacerbates Podocyte Injury in Diabetic Nephropathy by Inducing Cell Cycle Re‐entry and Inflammation Through Distinct Mechanisms Triggered by hnRNPU

Abstract Albuminuria is a hallmark of diabetic nephropathy (DN). Podocyte injury significantly contributes to proteinuria in DN. Our study found that lncRNA EVF‐2 is upregulated in podocytes of DN patients, correlating with cell cycle re‐entry and inflammation. Specific knockout or knockdown of lncRNA evf‐2 in diabetic mice or cultured podocytes alleviated podocyte injury associated with these processes. RNA sequencing of evf‐2‐overexpressing podocytes unveiled a predominant enrichment of upregulated mRNAs in cell cycle and inflammation pathways, with alternative splicing in cell cycle‐related mRNAs Ccnb1 and Tacc3. Chromatin isolation by RNA purification‐mass spectrometry (ChIRP‐MS) analysis highlighted the involvement of ribonucleoprotein complex and mRNA processing‐related proteins, with hnRNPU as the main binding partner of evf‐2 in spliceosomes. Knockdown of hnRNPU partially restored the upregulation of mRNAs induced by evf‐2 overexpression, altering splice variants of Ccnb1 and Tacc3. This study is the first to reveal the splice variants of cell cycle‐related genes in DN and elucidate the interaction between lncRNA evf‐2 and hnRNPU. This interaction culminates in the upregulation of cell cycle‐related genes and inflammatory factors through diverse pathways, potentially involving transcriptional activation, RNA stability modulation, alternative splicing or translational regulation. This highlights potential novel pathways for DN treatment.