Development of In Vitro Drug-Induced Cardiotoxicity Assay by Using Three-Dimensional Cardiac Tissues Derived from Human Induced Pluripotent Stem Cells

Abstract

An in vitro drug-induced cardiotoxicity assay is a critical step in drug discovery for clinical use. The use of human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) is promising for this purpose. However, single hiPSC-CMs are limited in their ability to mimic native cardiac tissue structurally and functionally, and the generation of artificial cardiac tissue using hiPSC-CMs is an ongoing challenging. We therefore developed a new method of constructing three-dimensional (3D) artificial tissues in a short time by coating extracellular matrix (ECM) components on cell surfaces. We hypothesized that 3D cardiac tissues derived from hiPSC-CMs (3D-hiPSC-CT) could be used for an in vitro drug-induced cardiotoxicity assay. 3D-hiPSC-CT were generated by fibronectin and gelatin nanofilm coated single hiPSC-CMs. Histologically, 3D-hiPSC-CT exhibited a sarcomere structure in the myocytes and ECM proteins, such as fibronectin, collagen type I/III, and laminin. The administration of cytotoxic doxorubicin at 5.0 μM induced the release of lactate dehydrogenase, while that at 2.0 μM reduced the cell viability. E-4031, human ether-a-go-go related gene (hERG)-type potassium channel blocker, and isoproterenol induced significant changes both in the Ca transient parameters and contractile parameters in a dose-dependent manner. The 3D-hiPSC-CT exhibited doxorubicin-sensitive cytotoxicity and hERG channel blocker/isoproterenol-sensitive electrical activity in vitro, indicating its usefulness for drug-induced cardiotoxicity assays or drug screening systems for drug discovery.

Mechanobiology Assays with Applications in Cardiomyocyte Biology and Cardiotoxicity

“…Several labs have deployed advanced video microscopy analyses of drug‐induced changes in contractile function of hiPSC‐CMs, from single cells to 3D tissue constructs. [ 91,98,108,114–116 ] Common mechanical readouts include changes in peak contraction/relaxation amplitudes, contraction/relaxation velocities, and beat rate (Table 2). For example, Sala et al deployed their “MUSCLEMOTION” analysis software to study the cardiotoxic agent Nifedipine (a Ca 2+ channel blocker used to treat hypertension) and learn that it reduces contraction amplitudes in single‐cell hiPSC‐CM, hiPSC‐CM monolayers, and 3D tissue constructs.…”

“…[ 143 ] In the past decade, there have been several studies that have reported changes in the contractile forces produced by EHTs in response to cardiotoxic agents identified by the CiPA initiative as well as cancer drugs using micropost displacements (Table 1). [ 144–149 ] For example, Hansen et al showed that the cardiotoxic agents quinidine, chromanol, erythromycin, and doxorubicin all negatively impacted CM function using EHTs. [ 149 ] With continued improvements in EHT micropost, their use is likely to grow in drug screening assays.…”

Building a new strategy for treating heart failure using Induced Pluripotent Stem Cells

“…2). Some reports have shown that cardiac constructs using iPS cells can indicate drug cytotoxicity in vitro [41], and that cardiac constructs with excessive fibrosis mimic diseased myocardium such as that seen in dilated cardiomyopathy, and can be used to analyze the pharmacological response to anti-fibrotic drugs in vitro.…”

Lipid availability influences the metabolic maturation of human pluripotent stem cell-derived cardiomyocytes

“…The regenerative capacity of human pluripotent stem cells (hPSCs) can provide a virtually unlimited source of cardiomyocytes (CMs) and other functional cells for toxicology and/or tissue engineering applications (Ye et al, 2014, Chong et al, 2014, Takeda et al, 2018. Recently developed cardiac differentiation protocols now generate CM purities approaching >90% without the use of growth factors in partially-or fully-defined conditions (Lian et al, 2012.…”