Abstract: PO0172
Blocking the Histone Lysine 79 Methyltransferase DOT1L Ameliorates AKI
Session Information
- AKI Mechanisms - 1
October 22, 2020 | Location: On-Demand
Abstract Time: 10:00 AM - 12:00 PM
Category: Acute Kidney Injury
- 103 AKI: Mechanisms
Authors
- Guan, Yingjie Angie, Rhode Island Hospital, Alpert Medical School of Brown University, Providence, Rhode Island, United States
- Zhuang, Shougang, Rhode Island Hospital, Alpert Medical School of Brown University, Providence, Rhode Island, United States
Background
The Disruptor of telomeric silencing 1-like (DOT1L) gene encodes a histone methyltransferase that methylates lysine-79 of histone H3 (H3K79). DOT1L-dependent H3K79 methylation plays an important role in various physiological and pathological processes, including transcriptional regulation, embryonic development and renal fibrosis. However, its role in acute kidney injury (AKI) tissue injury remains unknown.
Methods
AKI was induced in Male C57/B6 mice by bilateral ischemia-reperfusion (IR) injury or intraperitoneally injection with folic acid (FA). EPZ5676, a highly selective inhibitor of DOT1L (20 mg/kg) or an equal volume of vehicle was administered immediately after IRI or FA injection and then daily for two days. Renal function and histology were assessed by serum creatinine and HE staining. Immunohistochemistry and Western blotting were performed to identify tubule injury (NGAL), apoptosis (TUNEL and cleaved Caspase 3), proliferation (PCNA and Cyclin D) and Wnt signaling activation (active β-catenin and β-catenin).
Results
Mice developed AKI at 48 h after IR injury or FA administration as shown by the upregulation of serum creatinine and NGAL expression levels. Pharmacologic inhibition of DOT1L with EPZ5676 resulted in less severe tubular injury as evidenced by reduced renal dysfunction, diminished NGAL expression. Furthermore, the administration of EPZ5676 significantly reduced the number of TUNEL-positive and cleaved caspases-3 positive tubular cells in kidney tissues. Conversely, renal tubular cell proliferation was enhanced as indicated by increased expression of PCNA and cyclin D. Moreover, DOT1L inhibition by EPZ5676 up-regulated the expression of active β-catenin form and β -catenin.
Conclusion
Our data reveals that blocking DOT1L with EPZ5676 may protect against AKI in mice through inhibition of apoptosis and enhancement of kidney repair by a mechanism involved in the modulation of the canonical Wnt signaling pathway.
Funding
- Private Foundation Support