High-content screening of drug-induced cardiotoxicity using quantitative single cell imaging cytometry on microfluidic device.
Drug-induced cardiotoxicity or cytotoxicity followed by cell death in cardiac muscle is one of the major concerns in drug development. Herein, we report a high-content quantitative multicolor single cell imaging tool for automatic screening of drug-induced cardiotoxicity in an intact cell. A tunable multicolor imaging system coupled with a miniaturized sample platform was destined to elucidate drug-induced cardiotoxicity via simultaneous quantitative monitoring of intracellular sodium ion concentration, potassium ion channel permeability and apoptosis/necrosis in H9c2(2-1) cell line. Cells were treated with cisapride (a human ether-à-go-go-related gene (hERG) channel blocker), digoxin (Na(+)/K(+)-pump blocker), camptothecin (anticancer agent) and a newly synthesized anti-cancer drug candidate (SH-03). Decrease in potassium channel permeability in cisapride-treated cells indicated that it can also inhibit the trafficking of the hERG channel. Digoxin treatment resulted in an increase of intracellular [Na(+)]. However, it did not affect potassium channel permeability. Camptothecin and SH-03 did not show any cytotoxic effect at normal use (≤300 nM and 10 μM, respectively). This result clearly indicates the potential of SH-03 as a new anticancer drug candidate. The developed method was also used to correlate the cell death pathway with alterations in intracellular [Na(+)]. The developed protocol can directly depict and quantitate targeted cellular responses, subsequently enabling an automated, easy to operate tool that is applicable to drug-induced cytotoxicity monitoring with special reference to next generation drug discovery screening. This multicolor imaging based system has great potential as a complementary system to the conventional patch clamp technique and flow cytometric measurement for the screening of drug cardiotoxicity.
Research Institute of Pharmaceutical Sciences and College of Pharmacy, Seoul National University, Seoul, 151-742, South Korea.
Min Jung
MJ
Kim
Su Chul
SC
Lee
Sukdeb
S
Pal
Eunyoung
E
Han
Joon Myong
JM
Song
D000818:Animals; D000970:Antineoplastic Agents / Q000009:adverse effects / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D002316:Cardiotonic Agents / Q000494:pharmacology; D002460:Cell Line; D020117:Cisapride / Q000494:pharmacology; D004077:Digoxin / Q000494:pharmacology; D004353:Drug Evaluation, Preclinical / Q000295:instrumentation*; D004867:Equipment Design; D051638:Ether-A-Go-Go Potassium Channels / Q000037:antagonists & inhibitors; D006801:Humans; D019044:Image Cytometry / Q000295:instrumentation*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D032383:Myocytes, Cardiac / Q000187:drug effects*; D015221:Potassium Channels / Q000378:metabolism; D051381:Rats; D059010:Single-Cell Analysis / Q000295:instrumentation*; D012964:Sodium / Q000378:metabolism
D000970:Antineoplastic Agents; D002316:Cardiotonic Agents; D051638:Ether-A-Go-Go Potassium Channels; D015221:Potassium Channels; D004077:Digoxin; D012964:Sodium; D020117:Cisapride