Abstract: PO0638
Generation of Chimeric Nephrons in Newborn Mice for Testing Drug-Induced Nephrotoxicity
Session Information
- Development, Stem Cells, and Regenerative Medicine
November 04, 2021 | Location: On-Demand, Virtual Only
Abstract Time: 10:00 AM - 12:00 PM
Category: Development, Stem Cells, and Regenerative Medicine
- 500 Development, Stem Cells, and Regenerative Medicine
Authors
- Matsui, Kenji, Division of Nephrology and Hypertension, Department of Internal Medicine, The Jikei University School of Medicine, Tokyo, Japan
- Yamanaka, Shuichiro, Division of Nephrology and Hypertension, Department of Internal Medicine, The Jikei University School of Medicine, Tokyo, Japan
- Matsumoto, Naoto, Division of Nephrology and Hypertension, Department of Internal Medicine, The Jikei University School of Medicine, Tokyo, Japan
- Saito, Yatsumu, Division of Nephrology and Hypertension, Department of Internal Medicine, The Jikei University School of Medicine, Tokyo, Japan
- Takamura, Tsuyoshi, Division of Nephrology and Hypertension, Department of Internal Medicine, The Jikei University School of Medicine, Tokyo, Japan
- Fujimoto, Toshinari, Division of Nephrology and Hypertension, Department of Internal Medicine, The Jikei University School of Medicine, Tokyo, Japan
- Tajiri, Susumu, Division of Nephrology and Hypertension, Department of Internal Medicine, The Jikei University School of Medicine, Tokyo, Japan
- Matsumoto, Kei, Division of Nephrology and Hypertension, Department of Internal Medicine, The Jikei University School of Medicine, Tokyo, Japan
- Kobayashi, Eiji, Department of Kidney Regeneration, The Jikei University School of Medicine, Tokyo, Japan
- Yokoo, Takashi, Division of Nephrology and Hypertension, Department of Internal Medicine, The Jikei University School of Medicine, Tokyo, Japan
Background
Studies have reported certain limitations of experiments on animals for assessing nephrotoxicity. Due to species differences in tubular transporters, a model for evaluating nephrotoxicity in humans is desirable. Organoids are a novel research method, but the absence of their connection to the urinary tract makes them difficult for use in the evaluation of chronic nephrotoxicity.
Therefore, our goal was to develop a new pre-clinical assessment model that is amenable to repeated dose toxicity studies. In particular, this model aimed to produce human nephrons chimerized in the kidneys of host animals.
We previousely reported "exo utero" cell transplantation in which exogenous renal progenitor cells (RPCs) were transplanted into the retroperitoneal cavity of mouse fetuses without lethality. Although transplanted cells differentiated into functional nephrons chimerized with host kidneys, this method requires proficiency.
In this report, because continuous nephrogenesis takes place over several days after birth in mice, we used newborn mice as host animals and conducted a proof-of-concept study using mouse RPCs.
Methods
Metanephroi extracted from green fluorescent protein (GFP)-labeled mouse embryos were dissociated into single RPCs and injected into the subcapsular areas of newborn mice at postnatal days 0–1.
After 2 weeks, kidneys of host mice containing exogenous nephrons were extracted for the immunohistochemical evaluation. In addition, cisplatin was administered intraperitoneally to the host mice and the response of exogenous proximal tubular cells (PTCs) was evaluated.
Results
Immunohistochemistry revealed around 10% chimerism in glomeruli, proximal and distal tubules in the injected areas.
Exogenous PTCs exhibited dose-dependent expression of Kim-1 in response to cisplatin administration.
We aim to subsequently report data from single-cell RNA sequencing.
Conclusion
Mouse exogenous RPCs could form chimeric nephrons in newborn mice that reproduced drug-induced nephrotoxicity seen in native kidneys.
The neo-tubules are expected to be a tool that can be applied to long-term repetitive drug administration because of its integration into the host functioning nephrons.
Funding
- Government Support – Non-U.S.