Abstract: TH-PO1137
Downregulation of Renal Fibrosis Through Pyruvate Kinase M2 (PKM2) Inhibitor Compound 3k (3k) Regulation of CCN2 Expression
Session Information
- CKD: Mechanisms, AKI, and Beyond - 1
November 06, 2025 | Location: Exhibit Hall, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: CKD (Non-Dialysis)
- 2303 CKD (Non-Dialysis): Mechanisms
Authors
- Kosakai, Wakako, Saitama Ika Daigaku, Iruma, Saitama Prefecture, Japan
- Inoue, Tsutomu, Saitama Ika Daigaku, Iruma, Saitama Prefecture, Japan
- Watanabe, Yusuke, Saitama Ika Daigaku Kokusai Iryo Center, Hidaka, Saitama Prefecture, Japan
- Okada, Hirokazu, Saitama Ika Daigaku, Iruma, Saitama Prefecture, Japan
Background
Metabolic reprogramming and fibrosis are central to the progression of chronic kidney disease (CKD). However, the molecular mechanisms linking these processes are not yet fully understood. Our study demonstrates that a key glycolytic enzyme that catalyzes the final step of glycolysis, pyruvate kinase M2 (PKM2), serves as a key regulatory factor connecting metabolic reprogramming and fibrotic signaling.
Methods
A unilateral ischemia-reperfusion (IRI) mouse model was established using 6- to 8-week male C57/BL6 mice. The PKM2 inhibitor compound 3k, a PKM2-selective allosteric inhibitor derived from a novel naphthoquinone compound, was orally administered every other day (5 mg/kg BW) starting immediately after surgery, and the kidneys were harvested on day 14. Using the human renal tubular epithelial cell line (HK-2), we evaluated the expression levels and nuclear translocation of PKM2, YAP, TEAD, and β-catenin by RT-qPCR, Western blot, and immunoprecipitation (IP) after PKM2 knockdown by siRNA and 3k addition.
Results
Compared to the vehicle-treated control group, 3k group showed significantly reduced mRNA expression levels of CCN2 (0.241±0.103 [average of the vehicle-treated control=1.000]), collagen type1 (0.183±0.073), and fibronectin EIIIA (0.266±0.119), accompanied by histological improvement in fibrotic lesions as shown by Masson's trichrome staining. In HK-2 cells, the addition of siRNA_PKM2 or 3k reduced the expression of PKM2, YAP, TEAD and β- catenin, and inhibited their nuclear translocation compared with the control, accompanied by the
downregulation in CCN2 expression. In HK2 cells collected from the DMSO-treated control group with high CCN2 expression, co-precipitation of YAP and β-catenin was confirmed when anti-PKM2 antibody was used for IP.
Conclusion
A decrease in the nuclear translocation of PKM2 suppresses the Hippo-YAP-TEAD pathway and reduces the expression of the profibrotic factor, CCN2, thereby inhibiting the progression of renal fibrosis in the CKD mouse model. This study reveals the importance of PKM2 as a transcriptional regulator promoting renal fibrosis; and highlights the significance of PKM2 inhibition as a potential target for novel CKD therapeutics.
Funding
- Government Support – Non-U.S.