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FTO-mediated m6A modification alleviates diabetic nephropathy progression by downregulating the CYP2J3/Smurf2 axis
Yinhao Liu1, Chaohui Liu2, Yuqing Chen3
1Department of Diabetes Nephrology, Zhangzhou Traditional Chinese Medical Hospital, Zhangzhou, China.
Abstract:
The m6A demethylase FTO is implicated in epigenetic regulation, yet its specific mechanism in diabetic nephropathy (DN) remains unclear. This study investigates whether FTO mitigates DN progression by regulating the CYP2J3/Smurf2 axis. Mouse glomerular mesangial SV40-MES-13 cells were treated with high glucose (HG). Gene and protein expressions were assessed via qPCR and Western blot. MeRIP-qPCR and RIP assays evaluated m6A modifications and FTO-CYP2J3 mRNA binding. The CYP2J3-Smurf2 interaction was detected via immunofluorescence and co-Immunoprecipitation. Following FTO and CYP2J3 overexpression (OE), cell viability and fibrotic markers (p-Smad7/Smad7, TGF-β1, α-SMA, Fibronectin) were measured. In vivo validation was performed in DN mice, with evaluation of renal function alongside histological and immunohistochemical analyses. FTO was significantly downregulated in HG-treated cells. FTO directly interacted with CYP2J3 mRNA, and FTO OE decreased both the m6A modification level and the overall expression of CYP2J3. This reduction successfully suppressed the targeted binding between CYP2J3 and Smurf2. Functionally, FTO OE reversed HG-induced cell viability changes and inhibited the expression of downstream fibrotic proteins. Rescue experiments confirmed that CYP2J3 OE reversed the anti-fibrotic protective effects of FTO. In vivo, FTO OE significantly improved renal function parameters and attenuated pathological tissue injury and extracellular matrix accumulation in DN mice, effects which were counteracted by the co-overexpression of CYP2J3. FTO alleviates HG-induced renal fibrosis and overall DN progression by downregulating CYP2J3 via m6A modification, thereby reducing the CYP2J3-Smurf2 interaction.
Insights
The fat mass and obesity-associated protein (FTO) demethylase protects against diabetic nephropathy (DN) by downregulating CYP2J3 expression through m6A modification, thereby reducing kidney fibrosis.
Area of Science:
- Epigenetics
- Molecular Biology
- Nephrology
Background:
- The m6A demethylase FTO plays a role in epigenetic regulation, but its precise mechanism in diabetic nephropathy (DN) is not fully understood.
- Investigating FTO's role is crucial for understanding and potentially treating DN, a common complication of diabetes.
Purpose of the Study:
- To determine if FTO mitigates DN progression by regulating the CYP2J3/Smurf2 axis.
- To elucidate the molecular mechanisms underlying FTO's function in DN.
Main Methods:
- Utilized high glucose (HG)-treated mouse glomerular mesangial cells (SV40-MES-13) and in vivo DN mouse models.
- Assessed gene and protein expression via qPCR and Western blot; evaluated m6A modification and FTO-CYP2J3 mRNA binding using MeRIP-qPCR and RIP assays.
- Investigated CYP2J3-Smurf2 interaction via immunofluorescence and co-Immunoprecipitation; performed overexpression studies and functional rescue experiments.
Main Results:
- FTO was downregulated in HG-treated cells; FTO directly bound to CYP2J3 mRNA, reducing its m6A modification and expression.
- FTO overexpression suppressed CYP2J3-Smurf2 interaction, reversed HG-induced changes in cell viability, and inhibited fibrotic markers.
- In vivo, FTO overexpression improved renal function and attenuated kidney injury and extracellular matrix accumulation in DN mice.
Conclusions:
- FTO alleviates high glucose-induced renal fibrosis and DN progression by downregulating CYP2J3 via m6A modification.
- This mechanism involves reducing the CYP2J3-Smurf2 interaction, highlighting a novel therapeutic target for DN.
