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Combining Extracellular miRNA Determination with Microfluidic 3D Cell Cultures for the Assessment of Nephrotoxicity:

Laura Suter-Dick1, L Mauch2, D Ramp2

  • 1School of Life Sciences, University of Applied Sciences Northwestern Switzerland, Gründenstrasse 40, 4132, Muttenz, Switzerland. laura.suterdick@fhnw.ch.

The AAPS Journal
|July 25, 2018
PubMed

Insights

This study introduces a novel in vitro system using microfluidic cell cultures to detect drug-induced kidney injury. The findings highlight specific microRNAs as early and reliable biomarkers for renal proximal tubule damage.

Area of Science:

  • Nephrology
  • Toxicology
  • Biomarker Discovery

Background:

  • Drug-induced kidney injury (DIKI) is a significant clinical concern, potentially leading to long-term organ failure.
  • Current in vitro methods for detecting nephrotoxicity have limitations in predicting human outcomes.
  • There is a need for reliable and sensitive biomarkers to assess renal proximal tubule damage early.

Purpose of the Study:

  • To evaluate a novel in vitro system for detecting drug-induced renal proximal tubule damage.
  • To identify and validate microRNAs as sensitive biomarkers for early nephrotoxicity detection.
  • To assess the reproducibility and transferability of the developed in vitro system and biomarker panel.

Main Methods:

  • Implementation of organotypic cultures of human conditionally immortalized proximal tubule epithelial cells overexpressing organic anion transporter 1 (ciPTEC-OAT1) in a microfluidic OrganoPlate.
  • Exposure of ciPTEC-OAT1 cells to known nephrotoxicants: cisplatin, tenofovir, cyclosporine A, and tobramycin.
  • Measurement of cell viability (WST-8 assay), NAG release, and miRNA levels (mir-21, mir-29a, mir-34a, mir-192) in the culture medium.

Main Results:

  • MicroRNA levels in the culture medium served as earlier indicators of proximal tubule damage compared to cell viability and NAG release.
  • A panel of specific miRNAs (mir-29a, mir-34a, and mir-192) demonstrated high reproducibility across experiments and compounds.
  • The results were reproducible and technically transferable across two different laboratories, confirming the robustness of the system.

Conclusions:

  • The selected miRNAs function as sensitive biomarkers for damage to tubular epithelial cells induced by various nephrotoxicity mechanisms.
  • The 3D microfluidic culture system with ciPTEC-OAT1 cells and the miRNA biomarker panel offer a novel tool for in vitro nephrotoxicity detection.
  • This approach facilitates longitudinal, time-course in vitro toxicity studies, advancing the application of miRNAs in toxicology.
Keywords:
biomarkerkidneymiRNAmicrofluidicsnephrotoxicityorgan-on-a-chip

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