Related Experiment Video
Updated: Aug 10, 2026
Generation of Human Kidney Tubuloids from Tissue and Urine
Published on: April 16, 2021
A tubule cell model for ifosfamide nephrotoxicity
Katarina Aleksa1, Naomi Halachmi, Shinya Ito
1Division of Clinical Pharmacology and Toxicology, Hospital for Sick Children, Toronto, ON, Canada.
Abstract:
Mechanisms leading to ifosfamide (IF)-induced renal damage have not been fully elucidated. Recent work suggests that localized renal tubular metabolism of IF and the production of the nephrotoxic chloroacetaldehyde may lead to nephrotoxicity. Presently no pharmacological method to reduce IF nephrotoxicity has been identified. The objectives of this study were to establish a tubule cell model for IF nephrotoxicity, to verify whether renal proximal tubular cells have the necessary cytochrome P450 (CYP) enzymes to oxidize IF, and whether they can metabolize IF to chloroacetaldehyde. CYP3A, and 2B mRNA and protein were identified in LLCPK-1 cells. The cells metabolized the R- and S-IF enantiomers to their respective 2- and 3-dechloroethylifosfamide metabolites, by-products of chloroacetaldehyde formation. Metabolite production was both time and concentration-dependent. IF did not affect cell viability. In contrast, glutathione-depleted cells showed time and dose-dependent damage. The presence of the relevant CYP enzymes in renal tubular cells along with their ability to metabolize IF to its 2- and 3-dechloroethylifosfamide metabolites suggests that nephrotoxic damage may result from the localized production of chloroacetaldehyde. Glutathione is a major defence mechanism against IF toxicity, thus pharmacological methods for replenishing intracellular glutathione may be effective in modulating IF-induced nephrotoxicity.
Insights
Ifosfamide (IF) causes kidney damage through localized metabolism in renal tubules, producing toxic chloroacetaldehyde. Replenishing glutathione may protect against this ifosfamide-induced nephrotoxicity.
Area of Science:
- Nephrology
- Pharmacology
- Biochemistry
Background:
- Ifosfamide (IF) is a chemotherapy drug known to cause kidney damage.
- The precise mechanisms of IF-induced nephrotoxicity are not fully understood.
- Localized metabolism of IF in renal tubules and production of chloroacetaldehyde are suspected contributors.
Purpose of the Study:
- To create a renal tubule cell model for studying IF nephrotoxicity.
- To determine if renal proximal tubular cells possess the necessary cytochrome P450 (CYP) enzymes for IF metabolism.
- To investigate if these cells can metabolize IF into nephrotoxic chloroacetaldehyde.
Main Methods:
- Utilized LLCPK-1 cells as a tubule cell model.
- Detected CYP3A and CYP2B mRNA and protein expression.
- Analyzed the metabolism of R- and S-IF enantiomers and assessed cell viability.
- Investigated the role of glutathione depletion in IF toxicity.
Main Results:
- CYP3A and CYP2B enzymes were identified in LLCPK-1 cells.
- These cells metabolized IF enantiomers to dechloroethylifosfamide metabolites, indicating chloroacetaldehyde formation.
- Metabolite production was dependent on time and concentration.
- IF did not impact cell viability, but glutathione depletion led to significant cell damage.
Conclusions:
- Renal tubular cells possess the CYP enzymes capable of metabolizing IF.
- Localized production of chloroacetaldehyde in renal tubules likely contributes to IF-induced nephrotoxicity.
- Glutathione plays a critical role in defending against IF toxicity.
- Pharmacological strategies aimed at increasing intracellular glutathione levels may mitigate IF-induced kidney damage.
More Related Videos
Related Concept Videos
01:15Drug Elimination by Renal Route: Tubular Secretion
01:28Renal Drug Excretion: Tubular Secretion
01:29Acute Kidney Injury II: Pathophysiology