Abstract: FR-PO1056
HSP90 Inhibitor 17-DMAG Downregulated METTL3 and Attenuates Renal Fibrosis in AKI to CKD Model
Session Information
- CKD Mechanisms: Progression, Fibrosis, and Beyond
November 03, 2023 | Location: Exhibit Hall, Pennsylvania Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: CKD (Non-Dialysis)
- 2303 CKD (Non-Dialysis): Mechanisms
Authors
- Jin, Wencheng, Seoul National University, Gwanak-gu, Seoul, Korea (the Republic of)
- Shin, Nayeon, Seoul National University Seoul Metropolitan Government Boramae Medical Center, Dongjak-gu, Seoul, Korea (the Republic of)
- Weon, Boram, Seoul National University, Gwanak-gu, Seoul, Korea (the Republic of)
- Lee, Myoung Seok, Seoul National University Seoul Metropolitan Government Boramae Medical Center, Dongjak-gu, Seoul, Korea (the Republic of)
- Lee, Jeonghwan, Seoul National University, Gwanak-gu, Seoul, Korea (the Republic of)
Background
N6-methyladenosine (m6A) RNA methylation has been reported to participate in post-transcriptional gene expression regulation and development of kidney fibrosis.
Methods
AKI to CKD transition was induced by unilateral ischemic-reperfusion (uIRI) method (transient clamping followed by de-clamping of renal artery) in C57BL/6 mice. HSP90 Inhibitor 17-DMAG was injected intraperitoneally with 20 mg/kg dose. Renal fibrosis is evaluated using both imaging and molecular parameters. As imaging parameters, size, resistive index (RI), vascular index (VI; colored pixels / all pixels on microvascular imaging), and shear wave parameters were compared. Histology-based fibrosis score was also evaluated. And we evaluated the expression of fibrosis and inflammatory marker proteins such as TGF-β, α-SMA, Collagen 1 and E-Cadherin in uIRI model. Furthermore, we incubated human proximal tubular epithelial (HK2) cells under TGF-β (10 ng/ml) treatment as an in vitro model of kidney fibrosis.
Results
Imaging biomarkers of uIRI/17-DMAG showed better results than that of uIRI/vehicle (Size, 7.3±3.3mm vs. 10.7±0.86mm; RI, 0.79±0.09 vs. 0.68±0.05; VI, 9.4±6.0% vs. 18.0±4.1%, all p<0.05). Shear wave elastography did not show significant difference between uIRI/17-DMAG and uIRI/vehicle mice. 17-DMAG treatment attenuated the degrees of kidney fibrosis in vivo mice uIRI model (Histology-based fibrosis score 7.5±1.2 in uIRI/vehicle vs. 4.6±1.7 in uIRI/17-DMAG, P = 0.016). Expression of α-SMA and Collagen 1 proteins were lesser in uIRI/17-DMAG compared to uIRI/vehicle. In HK2 cell, 17-DMAG attenuated METTL3 expression regardless of TGFβ treatment. 17-DMAG treatment also downregulated epithelial marker (E-cadherin), and upregulated interstitial markers (N-cadherin, Vimentin, NET1). However, Hsp90 depletion using siHsp90RNA did not show METTL3 decrease in HK2 cell.
Conclusion
17-DMAG treatment attenuated renal fibrosis both in vitro TGFβ-challenged HK2 cell model and in vivo AKI to CKD mice model. METTL3 expression was diminished by 17-DMAG treatment.