Abstract: TH-PO0569
Establishment of a Drug Toxicity Assessment Using Kidney Organoids
Session Information
- Development, Stem Cells, and Regenerative Medicine
November 06, 2025 | Location: Exhibit Hall, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Development, Stem Cells, and Regenerative Medicine
- 600 Development, Stem Cells, and Regenerative Medicine
Authors
- Nakanoh, Hiroyuki, Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama, Japan
- Tsuji, Kenji, Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama, Japan
- Uchida, Naruhiko, Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama, Japan
- Fukushima, Kazuhiko, Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama, Japan
- Kitamura, Shinji, Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama, Japan
- Wada, Jun, Department of Nephrology, Rheumatology, Endocrinology and Metabolism, Okayama University Graduate School of Medicine, Dentistry, and Pharmaceutical Sciences, Okayama, Japan
Background
Efforts to reduce animal testing under the 3R principles (replacement, reduction and refinement) have highlighted new approach methodologies. Here, we developed a kidney organoid system derived from adult rat kidney stem (KS) cells and evaluated its utility as a screening platform for nephrotoxicity. Furthermore, we applied this system to assess the nephrotoxic effects of puberulic acid (PA), a compound suspected to be associated with acute kidney injury (AKI) cases linked to Beni-koji Cholestehelp supplements recently reported in Japan.
Methods
Nephrotoxicity was evaluated in KS cell-derived kidney organoids treated with known nephrotoxic agents, cisplatin and gentamicin for 72 hours. Assessment included histological analysis and qPCR analysis of kidney injury molecule-1 (Kim-1), a biomarker for AKI. Additionally, the nephrotoxicity of PA was investigated using both the kidney organoid model and an in vivo mouse model. C57BL/6N mice were administered PA intraperitoneally, and renal function and histopathological findings were evaluated after 4 days.
Results
In kidney organoids, treatment with cisplatin and gentamicin induced significant apoptosis, demonstrated by an increase in cleaved caspase-3–positive cells. Transmission electron microscopy revealed loss of intercellular junctions and cytoplasmic vacuolization, characterized as acute tubular necrosis (ATN). qPCR analysis showed a significant upregulation of Kim-1 expression in organoids treated with cisplatin and gentamicin. Similarly, exposure to PA resulted in proximal tubular injury in kidney organoids, accompanied by increased Kim-1 expression and a higher number of cleaved caspase-3–positive apoptotic cells. In vivo, PA-treated mice demonstrated impaired renal function, evidenced by elevated serum creatinine levels and an increased urinary albumin-to-creatinine ratio. Histological analysis revealed tubular atrophy, cell shedding and cast formation without glomerular abnormalities, consistent with ATN, paralleling the findings observed in organoids.
Conclusion
This study demonstrated the quantitative evaluation of renal tubular damage using kidney organoids and revealed the nephrotoxicity of PA in supplement-induced AKI. The KS cell-derived kidney organoid system offers a cost-effective, reproducible platform for nephrotoxicity screening as an alternative to animal models.
Funding
- Private Foundation Support