Nephrotoxicity Assessment with Human Kidney Tubuloids using Spherical Nucleic Acid-Based mRNA Nanoflares
Christian Wiraja1,2, Yutaro Mori1, Takaharu Ichimura1
1Division of Renal Medicine, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, United States.
Abstract:
Drug-induced nephrotoxicity represents an important cause of acute kidney injury with associated patient morbidity and mortality and is often responsible for termination of drug development, after extensive resource allocation. We have developed a human kidney tubuloid system that phenocopies, in 3D culture, kidney proximal tubules, a primary injury site of most nephrotoxicants. Traditional end point assays are often performed on 2D cultures of cells that have lost their differentiated phenotype. Herein, we pair a tubuloid system with Nanoflare (NF) mRNA nanosensors to achieve a facile, real-time assessment of drug nephrotoxicity. Using kidney injury molecule-1 (KIM-1) mRNA as a model injury biomarker, we verify NF specificity in engineered and adenovirus-transfected cells and confirm their efficacy to report tubular cell injury by aristolochic acid and cisplatin. The system also facilitates nephrotoxicity screening as demonstrated with 10 representative anticancer moieties. 5-Fluorouracil and paclitaxel induce acute tubular injury, as reflected by an NF signal increase.
Insights
A novel human kidney tubuloid system combined with Nanoflare mRNA nanosensors offers real-time assessment of drug-induced nephrotoxicity. This innovative approach aids in early detection of kidney injury from potential drug candidates.
Area of Science:
- Nephrology
- Biotechnology
- Toxicology
Background:
- Drug-induced nephrotoxicity is a significant cause of acute kidney injury, leading to patient morbidity, mortality, and drug development failure.
- Traditional toxicity assays use 2D cell cultures that often lack differentiated phenotypes, limiting their predictive power.
- Kidney proximal tubules are a primary site for drug-induced kidney injury.
Purpose of the Study:
- To develop a human kidney tubuloid system for phenocopying proximal tubules in 3D culture.
- To integrate Nanoflare (NF) mRNA nanosensors for real-time assessment of drug nephrotoxicity.
- To establish a facile screening platform for identifying nephrotoxic drug candidates.
Main Methods:
- Development of a 3D human kidney tubuloid culture system mimicking proximal tubules.
- Utilizing Nanoflare (NF) mRNA nanosensors for real-time detection of kidney injury biomarkers.
- Employing kidney injury molecule-1 (KIM-1) mRNA as a model biomarker for tubular cell injury.
- Screening of 10 anticancer agents for nephrotoxicity using the tubuloid-NF system.
Main Results:
- The human kidney tubuloid system successfully phenocopies proximal tubules.
- Nanoflare nanosensors demonstrated specificity and efficacy in reporting tubular cell injury.
- The system accurately detected nephrotoxicity induced by aristolochic acid and cisplatin.
- 5-Fluorouracil and paclitaxel were identified as inducers of acute tubular injury via NF signal increase.
Conclusions:
- The combined tubuloid and Nanoflare system provides a robust platform for real-time drug nephrotoxicity assessment.
- This model system accurately reflects drug-induced kidney injury in vitro.
- The platform facilitates efficient screening of drug candidates, potentially reducing late-stage development failures.
