Silencing of RAB3A Reshapes the Extracellular Vesicle ncRNA Landscape in Diabetic Podocytes and Uncovers miR-126-3p as a Candidate Mediator of Podocyte Injury

Diabetic nephropathy (DN) is a major cause of chronic kidney disease and end-stage renal failure, involving progressive damage to renal cells, including podocytes. Extracellular vesicles (EVs) have emerged as important mediators of intercellular communication in the kidney by transporting bioactive molecules such as non-coding RNAs. Rab3A, a small GTPase involved in vesicle docking and fusion at the plasma membrane, may influence EV biogenesis and the cargo composition. This study investigated the non-coding RNA content of EVs released by podocytes following RAB3A silencing under diabetic conditions. Cultured podocytes were transfected with RAB3A-specific siRNA and exposed to high glucose concentrations. EVs were isolated from culture media and characterized using complementary techniques, and their non-coding RNA cargo was analyzed via high-throughput sequencing and validated via RT-qPCR. Our results reveal that RAB3A silencing under high-glucose conditions induced significant changes in EV-associated non-coding RNAs, including altered levels of miR-126-3p. The context-dependent regulation of miR-126-3p suggests its involvement in IRS1/PI3K-Akt signaling, potentially affecting the balance between protective and pathogenic responses in the diabetic kidney. Overall, these findings provide new insights into the role of Rab3A in shaping EV cargo during diabetic stress and highlight EV-associated non-coding RNAs as potential contributors to the pathophysiology of DN.