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Microfluidic cell culture and its application in high-throughput drug screening: cardiotoxicity assay for hERG
Xiaojing Su1, Edmond W K Young, Heather A S Underkofler
1Department of Biomedical Engineering and Wisconsin Institutes for Medical Research, University of Wisconsin-Madison, Madison, WI, USA.
Abstract:
Evaluation of drug cardiotoxicity is essential to the safe development of novel pharmaceuticals. Assessing a compound's risk for prolongation of the surface electrocardiographic QT interval and hence risk for life-threatening arrhythmias is mandated before approval of nearly all new pharmaceuticals. QT prolongation has most commonly been associated with loss of current through hERG (human ether-a-go-go related gene) potassium ion channels due to direct block of the ion channel by drugs or occasionally by inhibition of the plasma membrane expression of the channel protein. To develop an efficient, reliable, and cost-effective hERG screening assay for detecting drug-mediated disruption of hERG membrane trafficking, the authors demonstrate the use of microfluidic-based systems to improve throughput and lower cost of current methods. They validate their microfluidics array platform in polystyrene (PS), cyclo-olefin polymer (COP), and polydimethylsiloxane (PDMS) microchannels for drug-induced disruption of hERG trafficking by culturing stably transfected HEK cells that overexpressed hERG (WT-hERG) and studying their morphology, proliferation rates, hERG protein expression, and response to drug treatment. Results show that WT-hERG cells readily proliferate in PS, COP, and PDMS microfluidic channels. The authors demonstrated that conventional Western blot analysis was possible using cell lysate extracted from a single microchannel. The Western blot analysis also provided important evidence that WT-hERG cells cultured in microchannels maintained regular (well plate-based) expression of hERG. The authors further show that experimental procedures can be streamlined by using direct in-channel immunofluorescence staining in conjunction with detection using an infrared scanner. Finally, treatment of WT-hERG cells with 5 different drugs suggests that PS (and COP) microchannels were more suitable than PDMS microchannels for drug screening applications, particularly for tests involving hydrophobic drug molecules.
Insights
This study introduces microfluidic systems for efficient hERG channel screening, crucial for assessing drug cardiotoxicity and preventing arrhythmias. Polystyrene and cyclo-olefin polymer microchannels proved superior for drug screening, especially with hydrophobic compounds.
Area of Science:
- Pharmacology
- Biotechnology
- Cardiovascular Research
Background:
- Drug-induced cardiotoxicity, particularly QT prolongation, poses a significant risk for life-threatening arrhythmias.
- The human ether-a-go-go related gene (hERG) potassium channel is a primary target for assessing drug cardiotoxicity.
- Current hERG screening methods can be costly and time-consuming, necessitating more efficient alternatives.
Purpose of the Study:
- To develop and validate a cost-effective, high-throughput microfluidic assay for screening drug-mediated disruption of hERG membrane trafficking.
- To evaluate the suitability of different microchannel materials (polystyrene, cyclo-olefin polymer, polydimethylsiloxane) for hERG screening.
Main Methods:
- Utilized microfluidic array platforms with stably transfected HEK cells overexpressing WT-hERG.
- Assessed cell morphology, proliferation, hERG protein expression via Western blot and immunofluorescence staining.
- Tested drug-induced disruption of hERG trafficking using 5 different drug compounds.
Main Results:
- WT-hERG cells proliferated effectively in polystyrene (PS), cyclo-olefin polymer (COP), and polydimethylsiloxane (PDMS) microchannels.
- Western blot analysis confirmed stable hERG protein expression in microchannel-cultured cells.
- PS and COP microchannels demonstrated greater suitability for drug screening, particularly for hydrophobic compounds, compared to PDMS.
Conclusions:
- Microfluidic systems offer an efficient and reliable platform for hERG screening assays.
- Polystyrene and cyclo-olefin polymer microchannels are promising for developing cost-effective cardiotoxicity screening tools.
- This approach can streamline pharmaceutical development by improving the assessment of drug-induced arrhythmias risk.
