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Steps for the Autologous Ex vivo Perfused Porcine Liver-kidney Experiment
Published on: December 18, 2013
A high-throughput liver-kidney metabolic interaction chip for insights into the nephrotoxicity mechanisms of
Siyu Liu1, Yun Yang1, Yifei Yang1
1Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, Beijing, 100700, China. syliu2024@163.com.
Abstract:
The kidney organ-on-a-chip (OoC) is a powerful tool for studying drug-induced nephrotoxicity, but its application is limited by the absence of liver metabolism and low throughput. Here, we developed a high-throughput liver-kidney OoC system (HLKOC) featuring microfluidics, plug-in biomimetic cups (MIMICups), scalable flow channel plates, precision-cut liver slices (PCLS), and 3D HK-2 cell spheroids. We first established a functional endothelial barrier by optimizing cell types, biomimetic blood flow rate, serum content, and membrane pore size. The structural and functional integrity of the PCLS and HK-2 spheroids within the MIMICups was then confirmed through histological staining, metabolic assays, and functional tests for viability, polarization, and transport. To evaluate the system's utility, we integrated the HLKOC with a single kidney OoC control and multidisciplinary techniques-including biochemical analysis, computational toxicology, molecular docking, metabolomics, and transcriptomics-to investigate the nephrotoxicity of triptolide (TPL) and its underlying mechanisms. Results showed that, compared to the single kidney OoC, the HLKOC exhibited higher levels of urea, total protein, and albumin in the biomimetic blood, confirming the robust biosynthetic capacity of the PCLS-based liver chip and its ability to better simulate in vivo conditions. Notably, a TPL-induced elevation in urea was observed only in the HLKOC, demonstrating the superior sensitivity of the liver-kidney co-culture. Multi-omics analysis revealed that TPL induced distinct metabolic and transcriptional responses in the HLKOC, involving pathways related to linoleic acid metabolism and vesicle-mediated processes, and led to the significant downregulation of transport proteins cubilin and GLUT1. These findings highlight the advantages of the HLKOC over single-organ systems for drug toxicity assessment and provide new insights into the mechanisms of TPL-induced nephrotoxicity.
Insights
A new high-throughput liver-kidney organ-on-a-chip (HLKOC) system enhances drug toxicity testing by incorporating liver metabolism. This advanced model reveals triptolide
Area of Science:
- Biomedical Engineering
- Toxicology
- Organ-on-a-Chip Technology
Background:
- Current kidney organ-on-a-chip (OoC) models lack liver metabolism, limiting their accuracy in assessing drug-induced nephrotoxicity.
- High-throughput screening is crucial for efficient drug development and toxicity evaluation.
Purpose of the Study:
- To develop and validate a high-throughput liver-kidney organ-on-a-chip (HLKOC) system.
- To investigate the nephrotoxicity of triptolide (TPL) and its mechanisms using the HLKOC system.
- To demonstrate the advantages of the HLKOC over single-kidney OoC models for drug toxicity assessment.
Main Methods:
- Development of a microfluidic HLKOC system integrating precision-cut liver slices (PCLS) and 3D kidney cell spheroids.
- Optimization of endothelial barrier function, liver and kidney tissue viability, and metabolic activity.
- Integration of multi-omics analyses (metabolomics, transcriptomics), biochemical assays, and computational toxicology to study TPL nephrotoxicity.
Main Results:
- The HLKOC system demonstrated robust liver biosynthetic capacity and simulated in vivo conditions more effectively than a single kidney OoC.
- Triptolide-induced urea elevation, indicative of nephrotoxicity, was detected exclusively in the HLKOC system, highlighting its superior sensitivity.
- Multi-omics analysis revealed TPL-induced alterations in linoleic acid metabolism, vesicle-mediated processes, and downregulation of key transport proteins (cubilin, GLUT1).
Conclusions:
- The high-throughput liver-kidney organ-on-a-chip (HLKOC) system offers enhanced sensitivity and accuracy for drug-induced nephrotoxicity assessment.
- The HLKOC system provides deeper mechanistic insights into drug toxicity compared to single-organ models.
- This advanced model holds significant potential for improving preclinical drug safety evaluations.
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