Kidney Week

Abstract: SA-PO0265

Endonuclease G-Mediated Apoptosis via Deoxyribonuclease (DNase) and Ribonuclease (RNase) Activity in Kidney Tubular Epithelial Cells

Session Information

• AKI: Mechanisms - Cell Signaling
  October 24, 2026 | Location: Exhibit Hall A, Convention Center
  Abstract Time: 10:00 AM - 12:00 PM

Category: Acute Kidney Injury

• 103 AKI: Mechanisms

Authors

• Li, Shenyang, Central Arkansas Veterans Healthcare System John L McClellan Memorial Veterans Hospital, Little Rock, Arkansas, United States

Shenyang Li, MD, PhD

• Wanchai, Visanu, University of Arkansas for Medical Sciences College of Medicine, Little Rock, Arkansas, United States

Visanu Wanchai, MSc, PhD

• Levurdiak, Olena, University of Arkansas for Medical Sciences College of Medicine, Little Rock, Arkansas, United States

Olena Levurdiak

• Shelton, Randal Shane, University of Arkansas for Medical Sciences College of Medicine, Little Rock, Arkansas, United States

Randal Shane Shelton, MS

• Fite, Todd, Central Arkansas Veterans Healthcare System John L McClellan Memorial Veterans Hospital, Little Rock, Arkansas, United States

Todd Fite

• Nookaew, Intawat, University of Arkansas for Medical Sciences College of Medicine, Little Rock, Arkansas, United States

Intawat Nookaew, PhD

• Basnakian, Alexei G., University of Arkansas for Medical Sciences College of Medicine, Little Rock, Arkansas, United States

Alexei G. Basnakian, MD, PhD

Background

Cisplatin injury to kidney tubular endothelial (KTE) cells is known to be mediated by apoptotic DNA endonucleases, particularly, endonuclease G (EndoG). It is highly induced by cisplatin, causes induction of other endonucleases and pre-mRNA alternative splicing and silencing of DNase I through its RNase activity that generates splice-switching oligonucleotides (SSO) from long non-coding RNA. However, other targets of EndoG-mediated alternative splicing and its role in KTE cells death remain unknown. We hypothesized that the alternative splicing induced by EndoG may occur in many other pro-apoptotic genes besides endonucleases.

Methods

To test this hypothesis, we overexpressed mature mouse EndoG tagged with cyan fluorescent protein (EndoG-O), or inhibited EndoG with our proprietary EndoG inhibitor PNR3-82 (EndoG-I), or both overexpressed and inhibited EndoG (EndoG-OI) in mouse tubular epithelial TKPTS cells. Cell death was measured using lactate dehydrogenase (LDH) release assay. RNA was isolated, converted to cDNA, barcoded, and sequenced on R9.4 FLO-MIN106D flow cells on a MinION instrument. Differential transcript usage (DTU) analysis was conducted to quantify condition specific changes in isoform composition independent of total gene expression.

Results

As expected, EndoG-OE caused significant cell death, while EndoG-I and EndoG-OI were protected from death. Correlation analysis demonstrated high within condition concordance and clear segregation between experimental groups. Hierarchical clustering further confirmed that EndoG perturbation induces robust transcriptomic remodeling. Across all pairwise comparisons, we identified 172 genes exhibiting significant Differential Transcript Usage (DTU) (p < 0.05). At least 12 apoptosis-related genes had transcript remodeling induced by overexpression or inhibition of EndoG.

Conclusion

These observations demonstrate a clear link between EndoG cytotoxicity and alternative splicing of various genes.

Funding

• Other NIH Support