Circ-0069561 as a novel diagnostic biomarker for progression of diabetic kidney disease.

BACKGROUND: Circular RNAs (circRNAs) are non-coding RNAs that are key regulators of the initiation and progression of various human diseases. However, the role of circRNAs in diabetic kidney disease (DKD) remains unknown. METHODS: Whole high-throughput RNA sequencing (RNA-seq) was performed on kidney tissues from clinical DKD patients and controls. Circ-0069561 with significantly up-regulated expression level was selected by real-time PCR (RT-PCR) analysis. RT-PCR and fluorescent in situ hybridization (FISH) further validated the expression and subcellular localization of circ-0069561 in type 2 diabetic mice and DKD patients. The clinical significance of circ-0069561 in DKD was evaluated. The circRNA-miRNA-ferroptosis associated mRNA network was constructed. The biological function of circ-0069561 in mouse podocyte clone 5 (MPC5) was analyzed. RESULTS: The top 10 up-regulated circular RNAs were selected by RT-PCR validation, and the results demonstrated a significant elevation in the expression level of circ-0069561. The RT-PCR and FISH results demonstrated that the expression of circ-0069561 was elevated in renal tissues of type 2 diabetic mice and DKD patients, with a predominant localization in glomerulus. The ROC curves showed that circ-0069561 had a good diagnostic value in massive proteinuria (area under the curve = 0.889). Kaplan-Meier analysis showed that high expression of circ-0069561 was associated with an increased risk of primary endpoints. The circRNA-miRNA-mRNA network indicated that ferroptosis might be involved in the pathogenesis of DKD. In vitro experiments demonstrated that circ-0069561 aggravated glucose-induced podocyte damage and ferroptosis. CONCLUSION: Circ-0069561 has the potential to be an ideal biomarker and therapeutic target for DKD progression.