Cardiotoxicity Hazard and Risk Characterization of ToxCast Chemicals Using Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes from Multiple Donors

Abstract

Heart disease remains a significant human health burden worldwide with a significant fraction of morbidity attributable to environmental exposures. However, the extent to which the thousands of chemicals in commerce and the environment may contribute to heart disease morbidity is largely unknown, because in contrast to pharmaceuticals, environmental chemicals are seldom tested for potential cardiotoxicity. Human induced pluripotent stem cell (iPSC)-derived cardiomyocytes have become an informative in vitro model for cardiotoxicity testing of drugs with the availability of cells from multiple individuals allowing in vitro testing of population variability. In this study, we hypothesized that a panel of iPSC-derived cardiomyocytes from healthy human donors can be used to screen for the potential cardiotoxicity hazard and risk of environmental chemicals. We conducted concentration−response testing of 1029 chemicals (drugs, pesticides, flame retardants, polycyclic aromatic hydrocarbons (PAHs), plasticizers, industrial chemicals, food/flavor/fragrance agents, etc.) in iPSC-derived cardiomyocytes from 5 donors. We used kinetic calcium flux and high-content imaging to derive quantitative measures as inputs into Bayesian population concentration−response modeling of the effects of each chemical. We found that many environmental chemicals pose a hazard to human cardiomyocytes in vitro with more than half of all chemicals eliciting positive or negative chronotropic or arrhythmogenic effects. However, most of the tested environmental chemicals for which human exposure and highthroughput toxicokinetics data were available had wide margins of exposure and, thus, do not appear to pose a significant human health risk in a general population. Still, relatively narrow margins of exposure (<100) were estimated for some perfuoroalkyl substances and phthalates, raising concerns that cumulative exposures may pose a cardiotoxicity risk. Collectively, this study demonstrated the value of using a population-based human in vitro model for rapid, high-throughput hazard and risk characterization of chemicals for which little to no cardiotoxicity data are available from guideline studies in animals.

Integrating nonlinear analysis and machine learning for human induced pluripotent stem cell‐based drug cardiotoxicity testing

“…The predicted IC50 values were 285.14 nM for isoproterenol, 142.97 nM for verapamil, and 1130.33 nM for cisapride (Figure 3a). Our IC50 values for isopropanol and verapamil were within the same range as previous studies (Heylman et al., 2015; Orvos et al., 2019), however our IC50 for cisapride was at the higher end of the wide range of reported values (Burnett et al., 2021b; Dempsey et al., 2016; Gilchrist et al., 2015; Sirenko et al., 2013). Embedding dimension proved to be an extremely effective and sensitive measure for calculating the inhibitory concentration necessary as it’s average measure was more sensitive than the benchmark beat frequency calculation.…”

“…In current study, the hiPSC‐CMs were used from tone donor (donor 1434, male). The hiPSC‐CMs from this donor have been well characterized in a number of publications (Blanchette et al., 2019; Burnett et al., 2021a; Martewicz et al., 2020), showing the ability to generate clinically‐relevant data. hiPSC‐CMs derived from female donors were shown to be more sensitive to hERG blockers, based on qualitative assessments (Zeng et al., 2019) and were more sensitive to drug‐induced QT prolongation (Huo et al., 2019).…”

“…The fluorescent signal of calcium transient was analyzed using ScreenWorks PeakPro software (Molecular Devices). Data from each timepoint was analyzed as detailed in previously published work (Burnett et al, 2019(Burnett et al, , 2021aSirenko et al, 2017). Here, we used "baseline" as a quantifying metric to ensure even beating from the samples in all the wells and across the plates prior to chemical treatment.…”

Acute effects of cardiac contractility modulation stimulation in conventional 2D and 3D human induced pluripotent stem cell-derived cardiomyocyte models

“…Conventional 2D monolayer human induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs), on stiff substrate, have been demonstrated to be useful for the evaluation of drugs and other chemical compounds ( Blinova et al, 2018 ; Burnett et al, 2021 ; Yang et al, 2022 ). However, such models may not be appropriate for medical device optimization or safety assessment where induced effects are intrinsically reliant on more complex culture conditions, which enable a 3D level tissue response (e.g., transmural lesion formation or physiological effects).…”

Integrating Physiologically Based Pharmacokinetic Modeling-Based Forward Dosimetry and in Vitro Bioassays to Improve the Risk Assessment of Organophosphate Esters on Human Health

“…The likelihood of an OPE inducing bioactivity in human tissues (RQ in-vitro , unitless) was calculated as the ratio of the predicted concentration in the target tissue using PBPK-based forward dosimetry to the lowest POD-inducing bioactivity in the corresponding tissue (eq S33). 41 ■ RESULTS AND DISCUSSION Cumulative Exposure Assessment of OPEs. As listed in Tables S1−S9, the total concentrations of OPEs (∑OPEs) in soil, water, and food in China were in the range of 4.39−422 ng/g (mean ± SD: 61.9 ± 81.3 ng/g), 8.54−213 ng/L (77.8 ± 66.4 ng/L), and 0.205−72.7 ng/g ww (9.61 ± 14.4 ng/g ww), respectively, which were comparable to or higher than those in other countries, such as with tap water in Korea (53.0 ng/L) and cereals in Sweden (2.2 ng/g dw), Australia (2.03 ng/g dw), and the U.S. (not detected).…”