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Updated: Mar 21, 2026
High-Throughput Cardiotoxicity Screening Using Mature Human Induced Pluripotent Stem Cell-Derived Cardiomyocyte Monolayers
Published on: March 24, 2023
Modeling Doxorubicin-Induced Cardiotoxicity in Human Pluripotent Stem Cell Derived-Cardiomyocytes
Agnes Maillet1, Kim Tan1, Xiaoran Chai2
1Translational Laboratory in Genetic Medicine, National University of Singapore and the Agency for Science Technology and Research (A*STAR), Singapore.
Abstract:
Doxorubicin is a highly efficacious anti-cancer drug but causes cardiotoxicity in many patients. The mechanisms of doxorubicin-induced cardiotoxicity (DIC) remain incompletely understood. We investigated the characteristics and molecular mechanisms of DIC in human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs). We found that doxorubicin causes dose-dependent increases in apoptotic and necrotic cell death, reactive oxygen species production, mitochondrial dysfunction and increased intracellular calcium concentration. We characterized genome-wide changes in gene expression caused by doxorubicin using RNA-seq, as well as electrophysiological abnormalities caused by doxorubicin with multi-electrode array technology. Finally, we show that CRISPR-Cas9-mediated disruption of TOP2B, a gene implicated in DIC in mouse studies, significantly reduces the sensitivity of hPSC-CMs to doxorubicin-induced double stranded DNA breaks and cell death. These data establish a human cellular model of DIC that recapitulates many of the cardinal features of this adverse drug reaction and could enable screening for protective agents against DIC as well as assessment of genetic variants involved in doxorubicin response.
Insights
Doxorubicin causes heart damage (cardiotoxicity) through poorly understood mechanisms. This study uses human stem cell-derived heart cells to model cardiotoxicity and identify TOP2B as a key factor in doxorubicin-induced DNA damage and cell death.
Area of Science:
- Cardiology
- Molecular Biology
- Genetics
Background:
- Doxorubicin is a potent anti-cancer drug with known cardiotoxicity.
- The precise mechanisms underlying doxorubicin-induced cardiotoxicity (DIC) are not fully elucidated.
- Understanding DIC is crucial for patient safety during chemotherapy.
Purpose of the Study:
- To investigate the molecular mechanisms and characteristics of DIC.
- To establish a human cellular model for studying DIC.
- To identify potential therapeutic targets for mitigating doxorubicin cardiotoxicity.
Main Methods:
- Utilized human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) as a model system.
- Assessed doxorubicin effects on cell viability, reactive oxygen species, mitochondrial function, and intracellular calcium.
- Employed RNA-sequencing (RNA-seq) for genome-wide gene expression analysis.
- Used multi-electrode array (MEA) technology to evaluate electrophysiological changes.
- Applied CRISPR-Cas9 gene editing to disrupt the TOP2B gene.
Main Results:
- Doxorubicin induced dose-dependent cell death, oxidative stress, mitochondrial dysfunction, and altered calcium handling in hPSC-CMs.
- RNA-seq revealed significant changes in gene expression patterns following doxorubicin treatment.
- MEA analysis identified doxorubicin-induced electrophysiological abnormalities.
- CRISPR-Cas9-mediated knockout of TOP2B reduced hPSC-CM sensitivity to doxorubicin-induced DNA damage and cell death.
Conclusions:
- Established a robust human cellular model of doxorubicin-induced cardiotoxicity (DIC).
- Demonstrated the critical role of TOP2B in doxorubicin-induced DNA damage and cell death in human cardiomyocytes.
- The model provides a platform for screening protective agents and assessing genetic factors influencing doxorubicin response.
