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Updated: Aug 6, 2025
Use of Enzymatic Biosensors to Quantify Endogenous ATP or H2O2 in the Kidney
Published on: October 12, 2015
ATP/ADP biosensor organoids for drug nephrotoxicity assessment
Koichiro Susa1,2,3, Kenichi Kobayashi2,4, Pierre Galichon1,2
1Renal Division, Department of Medicine, Brigham and Women's Hospital, Boston, MA, United States.
Abstract:
Drug nephrotoxicity is a common healthcare problem in hospitalized patients and a major limitation during drug development. Multi-segmented kidney organoids derived from human pluripotent stem cells may complement traditional cell culture and animal experiments for nephrotoxicity assessment. Here we evaluate the capability of kidney organoids to investigate drug toxicity in vitro. Kidney organoids express renal drug transporters, OAT1, OAT3, and OCT2, while a human proximal tubular cell line shows the absence of OAT1 and OAT3. Tenofovir and aristolochic acid (AA) induce proximal tubular injury in organoids which is ameliorated by an OAT inhibitor, probenecid, without damage to podocytes. Similarly, cisplatin causes proximal tubular damage that can be relieved by an OCT inhibitor, cimetidine, collectively suggesting the presence of functional OATs and OCTs in organoid proximal tubules. Puromycin aminonucleoside (PAN) induced segment-specific injury in glomerular podocytes in kidney organoids in the absence of tubular injury. Reporter organoids were generated with an ATP/ADP biosensor, which may be applicable to high-throughput screening in the future. In conclusion, the kidney organoid is a useful tool for toxicity assessment in the multicellular context and may contribute to nephrotoxicity assessment during drug development.
Insights
Kidney organoids effectively model drug-induced kidney injury, showing functional transporters and segment-specific toxicity. This tool aids in evaluating drug nephrotoxicity during development.
Area of Science:
- Nephrology
- Toxicology
- Stem Cell Biology
Background:
- Drug nephrotoxicity poses significant clinical and developmental challenges.
- Traditional methods like cell cultures and animal models have limitations in predicting human kidney toxicity.
- Human pluripotent stem cell-derived kidney organoids offer a promising in vitro alternative.
Purpose of the Study:
- To evaluate the utility of kidney organoids for in vitro drug nephrotoxicity assessment.
- To investigate the expression and function of renal drug transporters in kidney organoids.
- To determine the capacity of kidney organoids to model segment-specific drug-induced kidney injury.
Main Methods:
- Generation of multi-segmented kidney organoids from human pluripotent stem cells.
- Assessment of renal drug transporter expression (OAT1, OAT3, OCT2) using quantitative methods.
- Exposure of kidney organoids to nephrotoxic drugs (tenofovir, aristolochic acid, cisplatin, puromycin aminonucleoside).
- Evaluation of drug-induced injury in specific kidney segments (proximal tubules, podocytes).
- Utilizing specific inhibitors (probenecid, cimetidine) to confirm transporter involvement.
- Development of reporter organoids with an ATP/ADP biosensor for potential high-throughput screening.
Main Results:
- Kidney organoids express key renal drug transporters (OAT1, OAT3, OCT2), unlike a human proximal tubular cell line lacking OAT1/OAT3.
- Tenofovir and aristolochic acid induced proximal tubular injury, ameliorated by an OAT inhibitor (probenecid).
- Cisplatin caused proximal tubular damage, relieved by an OCT inhibitor (cimetidine), confirming functional transporters.
- Puromycin aminonucleoside induced segment-specific podocyte injury without tubular damage.
- Reporter organoids with an ATP/ADP biosensor were successfully generated.
Conclusions:
- Kidney organoids accurately model drug-induced proximal tubular and podocyte injury.
- Functional renal drug transporters are present in kidney organoid proximal tubules.
- Kidney organoids serve as a valuable in vitro tool for assessing drug nephrotoxicity in a multicellular context.
- This model holds potential for improving drug development and safety assessment.
