An impedance-based cellular assay using human iPSC-derived cardiomyocytes to quantify modulators of cardiac
Clay W Scott1, Xiaoyu Zhang2, Najah Abi-Gerges2
1*Drug Safety and Metabolism, AstraZeneca Pharmaceuticals, Waltham, Massachusetts 02451, ACEA Biosciences Inc., San Diego, California 92121 and Drug Safety and Metabolism, AstraZeneca Pharmaceuticals, Alderley Park, SK10 4TG, UK clay.scott@astrazeneca.com.
Abstract:
Cardiovascular toxicity, a prominent reason for late-stage failures in drug development, has resulted in a demand for in vitro assays that can predict this liability in early drug discovery. Current in vitro cardiovascular safety testing primarily focuses on ion channel modulation and low throughput cardiomyocyte (CM) contractility measurements. We evaluated both human induced pluripotent stem cell-derived CMs (hiPSC-CMs) and rat neonatal CMs (rat CMs) on the xCELLigence Cardio system which uses impedance technology to quantify CM beating properties in a 96-well format. Forty-nine compounds were tested in concentration-response mode to determine potency for modulation of CM beating, a surrogate biomarker for contractility. These compounds had previously been tested in vivo and in a low throughput in vitro optical-based contractility assay that measures sarcomere shortening in electrically paced dog CMs. In comparison with in vivo contractility effects, hiPSC-CM impedance had assay sensitivity, specificity, and accuracy values of 90%, 74%, and 82%, respectively. These values compared favorably to values reported for the dog CM optical assay (83%, 84%, and 82%) and were slightly better than impedance using rat CMs (77%, 74%, and 74%). The potency values from the hiPSC-CM and rat CM assays spanned four orders of magnitude and correlated with values from the dog CM optical assay (r(2 )= 0.76 and 0.70, respectively). The Cardio system assay has >5× higher throughput than the optical assay. Thus, hiPSC-CM impedance testing can help detect the human cardiotoxic potential of novel therapeutics early in drug discovery, and if a hazard is identified, has sufficient throughput to support the design-make-test-analyze cycle to mitigate this liability.
Insights
This study introduces a high-throughput impedance assay using human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) to predict drug-induced cardiovascular toxicity early in drug discovery, improving upon existing methods.
Area of Science:
- Drug discovery and development
- Cardiovascular toxicology
- Stem cell biology
Background:
- Cardiovascular toxicity is a major cause of late-stage drug development failures.
- Current in vitro cardiovascular safety assays have limitations in throughput and predictive power.
- There is a need for reliable early-stage in vitro assays to predict cardiotoxicity.
Purpose of the Study:
- To evaluate the utility of the xCELLigence Cardio system using impedance technology for assessing drug-induced cardiotoxicity.
- To compare the performance of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) and rat neonatal cardiomyocytes (rat CMs) in this assay.
- To determine if the impedance assay can accurately predict in vivo cardiotoxicity and correlate with existing in vitro assays.
Main Methods:
- Human induced pluripotent stem cell-derived CMs (hiPSC-CMs) and rat neonatal CMs (rat CMs) were cultured on the xCELLigence Cardio system (96-well format).
- Forty-nine compounds were tested in concentration-response to measure modulation of CM beating, a surrogate for contractility.
- Assay performance was compared against in vivo data and a low-throughput dog CM optical contractility assay.
Main Results:
- The hiPSC-CM impedance assay demonstrated high sensitivity (90%), specificity (74%), and accuracy (82%) in predicting in vivo cardiotoxicity.
- These performance metrics favorably compared to the dog CM optical assay and outperformed the rat CM impedance assay.
- Potency values from hiPSC-CM and rat CM impedance assays correlated well with the dog CM optical assay (r²=0.76 and 0.70, respectively).
- The Cardio system assay offers >5x higher throughput compared to the optical assay.
Conclusions:
- hiPSC-CM impedance testing is a valuable tool for early detection of human cardiotoxic potential in novel therapeutics.
- The assay's high throughput supports iterative drug design and development cycles for mitigating cardiotoxicity liabilities.
- This impedance-based assay provides a more efficient and predictive method for cardiovascular safety assessment in drug discovery.
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