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Updated: Jan 13, 2026
Isolation of Primary Human Proximal Tubule Epithelial Cells and Their Use in Creating a Microphysiological Model of the Renal Proximal Tubule
Published on: May 9, 2025
Predicting nucleic acid drug-induced nephrotoxicity using a 3D human renal proximal tubule spheroid model
Kaoru Morimura1,2, Etsushi Takahashi1, Hayata Maeda1
1Bio Business Promotion Department, Medical Division, NIKKISO Co., Ltd.
Abstract:
Nucleic acid drugs hold considerable promise; however, their toxicological profiles are often difficult to assess in animal models. Clinical studies have reported adverse effects, including thrombocytopenia, complement activation, hepatotoxicity, and nephrotoxicity. While human cell-based models for hepatotoxicity are advancing, nephrotoxicity assessment remains limited by the scarcity of physiologically relevant kidney cells. In this study, a three-dimensional spheroid model of human primary renal proximal tubule epithelial cells (3D-RPTEC, Nikkiso) was employed to evaluate the nephrotoxicity of nucleic acid drugs. Proteomic profiling revealed enhanced expression of drug transporters and endocytic machinery in 3D-RPTEC compared with two-dimensional cultures. Lipofection enabled efficient intracellular delivery of nucleic acids. Toxicity was assessed using ATP quantification, biomarker analysis (LDH, KIM-1, NGAL), and high-content analysis (HCA). Significant ATP depletion was observed only after prolonged exposure to SPC5001, a nephrotoxic antisense oligonucleotide. In contrast, biomarker expression and HCA facilitated early detection of compound-specific toxicity and implicated endoplasmic reticulum and mitochondrial stress as underlying mechanisms. These findings establish 3D-RPTEC as a sensitive and physiologically relevant platform for predicting the nephrotoxic potential of nucleic acid drugs.
Insights
Assessing nucleic acid drug toxicity in kidneys is challenging. A new 3D human kidney cell model (3D-RPTEC) effectively predicts drug-induced nephrotoxicity, enabling earlier and more accurate safety assessments.
Area of Science:
- Pharmacology
- Toxicology
- Cell Biology
Background:
- Nucleic acid drugs offer therapeutic promise but pose toxicological challenges, particularly nephrotoxicity.
- Assessing drug-induced kidney damage is difficult due to limited availability of relevant human kidney cells.
- Existing models struggle to accurately predict the nephrotoxic potential of novel therapeutics.
Purpose of the Study:
- To develop and validate a physiologically relevant human cell model for assessing nucleic acid drug nephrotoxicity.
- To evaluate the utility of a three-dimensional renal proximal tubule epithelial cell (3D-RPTEC) model for predicting kidney toxicity.
- To compare the sensitivity of different toxicity assessment methods in the 3D-RPTEC model.
Main Methods:
- Utilized a three-dimensional spheroid model of human primary renal proximal tubule epithelial cells (3D-RPTEC).
- Employed proteomic profiling to characterize cell function and lipofection for nucleic acid delivery.
- Assessed toxicity via ATP quantification, biomarker analysis (LDH, KIM-1, NGAL), and high-content analysis (HCA).
Main Results:
- 3D-RPTEC cultures exhibited enhanced expression of drug transporters and endocytic machinery compared to 2D cultures.
- Early detection of compound-specific toxicity was achieved using biomarker analysis and HCA.
- Endoplasmic reticulum and mitochondrial stress were identified as key mechanisms underlying observed nephrotoxicity.
Conclusions:
- The 3D-RPTEC spheroid model is a sensitive and physiologically relevant platform for evaluating nucleic acid drug nephrotoxicity.
- This model facilitates early identification of potential kidney toxicity, improving drug safety assessment.
- Biomarker analysis and HCA in the 3D-RPTEC model provide mechanistic insights into drug-induced cellular stress.
