Abstract: FR-PO249
An ATP/ADP Biosensor as a Real-Time Toxicity Assay in Kidney Organoids
Session Information
- Stem Cells
November 03, 2017 | Location: Hall H, Morial Convention Center
Abstract Time: 10:00 AM - 10:00 AM
Category: Developmental Biology and Inherited Kidney Diseases
- 402 Stem Cells
Authors
- Galichon, Pierre, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States
- Susa, Koichiro, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States
- Gupta, Navin R., Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States
- Bonventre, Joseph V., Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States
- Morizane, Ryuji, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts, United States
Background
Renal toxicity is frequent and a major limitation to drug development. Renal organoids have been generated from human pluripotent stem cells (hPSCs), overcoming limitations related to the expense and translatability of cell culture and animal experiments. Here, we present a novel system to evaluate the toxicity of drugs with a real-time biosensor of ATP/ADP ratio in kidney organoids.
Methods
Stable hPSC lines, expressing the ATP/ADP ratio PercevalHR, were established with lentiviral transduction and differentiated in kidney organoids as previously described. Organoids were used fresh or after freeze/thaw. ATP and ADP signal were acquired via live fluorescence microscopy, and a ratio was quantified with the ImageJ RatioPlus plugin. Organoids were exposed to various toxicants (10 or 50 µM cisplatin, 10 mg/ml aristolochic acid (AA), 5 mM sodium azide (NaN3) or vehicle.
Results
The ATP and ADP signals were greater in nephron epithelial structures than in the stromal compartment and NaN3 induced a rapid decrease in ATP/ADP ratio within one minute. Cisplatin and AA significantly reduced the ATP/ADP ratio as compared to control organoids after 15 hours. Reduction in ATP/ADP ratios preceded contraction of the organoids.
Conclusion
Reduction in ATP/ADP ratio in kidney organoids is a sensitive indicator of drug toxicity, offering a direct live readout in human tissues in vitro. The unlimited availability of hPSC-derived organoids and the ability to freeze them at the pre-analytical stage is well-suited to high-throughput screening and might facilitate the identification of lead candidates during drug discovery.
A: ATP/ADP ratio imaging in a thawed organoid
B: Mean ATP/ADP ratio in organoids exposed to nephrotoxicants (n=6/group)
Funding
- Private Foundation Support