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Kazuhiro Hasegawa1, Yusuke Sakamaki2, Masanori Tamaki1
1Department of Nephrology, Tokushima University Graduate School of Biomedical Sciences, 3-18-15 Kuramoto-Cho, Tokushima 770-8503, Japan.
Abstract:
Acute kidney injury (AKI) is a major complication of lupus nephritis and kidney transplantation, inevitably causing ischemia-reperfusion (I/R) injury. We previously confirmed that high-dose voclosporin induces drug nephropathy through aberrant peroxisome accumulation. The latter induces increased renal indole-3-aceticT acid (IAA) production due to the decreased expression of the IAA-degrading enzyme indolethylamine N-methyltransferase (INMT). Conversely, INMT overexpression prevents this nephropathy, suggesting that high-dose voclosporin could enable a novel therapeutic approach. This prompted us to test whether INMT overexpression with high-dose voclosporin could avert nephrotoxicity and protect against I/R injury. Inmt-overexpressing mice treated with high-dose voclosporin exhibited absence of peroxisomal abnormalities and resistance to I/R injury. RNA sequencing revealed the downregulation of tubular injury markers NGAL (Lcn2) and KIM-1 (Havcr1) concurrent with significant cytokine suppression. Mechanistic analysis revealed the robust induction of Regnase-2, an mRNA decay factor, which directly targeted stem-loop structures within the 3' untranslated region of Lcn2 and Havcr1, thereby promoting their degradation in proximal tubular cells. Importantly, Regnase-2 knockdown mice showed Lcn2 upregulation, mitochondrial dysfunction, and peroxisomal abnormalities culminating in AKI, underscoring its renal protective effects. High-dose voclosporin under Inmt overexpression promoted Regnase-2-mediated mRNA decay to suppress tubular injury. This protective effect extended beyond I/R to rhabdomyolysis- and lipopolysaccharide-induced AKI to prevent nephropathy. Our findings demonstrate the potential transformative therapeutic approach of administering high-dose voclosporin to promote the prophylactic effect of Regnase-2 augmentation against AKI in both native and transplanted human kidneys.
Insights
Overexpressing indolethylamine N-methyltransferase (INMT) with high-dose voclosporin prevents acute kidney injury (AKI) by enhancing Regnase-2. This approach protects against ischemia-reperfusion and other AKI types.
Area of Science:
- Nephrology
- Molecular Biology
- Immunology
Background:
- Acute kidney injury (AKI) is a significant complication in lupus nephritis and kidney transplantation, often involving ischemia-reperfusion (I/R) injury.
- High-dose voclosporin can induce nephropathy via peroxisome accumulation, leading to increased indole-3-acetic acid (IAA) due to reduced indolethylamine N-methyltransferase (INMT) expression.
- Previous findings suggest INMT overexpression can prevent voclosporin-induced nephropathy.
Purpose of the Study:
- To investigate if INMT overexpression combined with high-dose voclosporin can prevent nephrotoxicity and protect against I/R injury.
- To elucidate the underlying molecular mechanisms of this protective effect.
Main Methods:
- Utilized INMT-overexpressing mice treated with high-dose voclosporin.
- Assessed peroxisomal abnormalities and resistance to I/R injury.
- Performed RNA sequencing to analyze tubular injury markers (NGAL/Lcn2, KIM-1/Havcr1) and cytokine expression.
- Investigated the role of Regnase-2 in mRNA decay of injury markers.
- Examined AKI models induced by I/R, rhabdomyolysis, and lipopolysaccharide.
Main Results:
- INMT-overexpressing mice treated with high-dose voclosporin showed no peroxisomal abnormalities and were resistant to I/R injury.
- RNA sequencing revealed downregulation of NGAL (Lcn2) and KIM-1 (Havcr1) and suppressed cytokines.
- Regnase-2 was robustly induced and directly targeted 3' UTR stem-loop structures of Lcn2 and Havcr1, promoting their degradation.
- Regnase-2 knockdown led to Lcn2 upregulation, mitochondrial dysfunction, and AKI.
- The protective effect extended to rhabdomyolysis- and lipopolysaccharide-induced AKI.
Conclusions:
- High-dose voclosporin with INMT overexpression promotes Regnase-2-mediated mRNA decay, suppressing tubular injury and preventing AKI.
- Regnase-2 plays a critical renal protective role against various AKI insults.
- This strategy offers a potential therapeutic approach for preventing AKI in native and transplanted kidneys.
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