METTL3/YTHDF3 m⁶A axis promotes ferroptosis in diabetic kidney disease by stabilizing TfR1

Diabetic kidney disease (DKD) is a common complication of diabetes. N6-Methyladenosine (m⁶A) modification is a widely studied epigenetic mechanism. Methyltransferase-like (METTL) 3 is a well-studied methyltransferase. This study aimed to investigate the role of METTL3 in DKD and the underlying mechanism.

Thirty-five DKD patients and 28 control volunteers were recruited. Animal and cell DKD models were established. QRT-PCR and Western blot were performed to analyze the expression of METTL3 and fibrosis-related indicators. Cell viability and proliferation were assessed via a cell counting kit-8 and colony formation assays. Ferrous iron (Fe²⁺), malonaldehyde (MDA), and glutathione (GSH) contents were measured by commercial kits. The interaction between METTL3/YTH N6-methyladenosine RNA binding protein (YTHDF)3 and transferrin receptor-1 (TfR1) was examined through RNA immunoprecipitation and dual-luciferase reporter assays.

Results showed that METTL3-mediated m⁶A modification was elevated in kidney tissues of DKD patients and in high glucose (HG)-treated human renal mesangial cells (HRMCs). Besides, HG-treated HRMCs showed increased ferroptosis. In addition, METTL3 inhibition increased cell proliferation and inhibited ferroptosis in HRMCs. Mechanically, the METTL3/YTHDF3 m⁶A axis enhanced the stability of TfR1 mRNA. Moreover, YTHDF3 inhibition increased cell proliferation and inhibited ferroptosis in HRMCs. Finally, in vivo study results indicated that METTL3 deficiency inhibited ferroptosis and improved pathological damages.

In summary, METTL3/YTHDF3 m⁶A axis promoted ferroptosis in DKD by stabilizing TfR1, which could provide a reference for DKD treatment.