2018 Progress Report: High-throughput Hazard,Dose-responseandPopulationVariabilityAssessmentof Cardiotoxicity in aHumanInducedPluripotentStem Cell(iPSC)-derivedin vitro Culture Model

EPA Grant Number: R835802C001
Subproject: this is subproject number 001 , established and managed by the Center Director under grant R835802
(EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).

Center: Organotypic Culture Models For Predictive Toxicology Center
Center Director: Rusyn, Ivan
Title: High-throughput Hazard,Dose-responseandPopulationVariabilityAssessmentof Cardiotoxicity in aHumanInducedPluripotentStem Cell(iPSC)-derivedin vitro Culture Model
Investigators: Rusyn, Ivan , Threadgill, David W. , Wright, Fred A.
Institution: Texas A & M University
EPA Project Officer: Klieforth, Barbara I
Project Period: June 1, 2015 through May 31, 2019 (Extended to May 31, 2020)
Project Period Covered by this Report: June 1, 2018 through May 31,2019
RFA: Organotypic Culture Models for Predictive Toxicology Center (2013) RFA Text |  Recipients Lists
Research Category: Human Health , Health , Safer Chemicals

Objective:

Important target areas for addressing data gaps through in vitro screening include evaluations of cardiotoxicity. Despite the fact that current conservative estimates relate at least 23% of all cardiovascular disease cases to environmental exposures, the identities of the causative environmental agents remain largely uncharacterized. Moreover, cardiotoxicity remains among the most pronounced reasons, comparable to those associated with hepatotoxicity, for drug attrition during clinical trials and post-marketing. Current pre-clinical approaches for cardiophysiologic evaluations of chemicals rely almost exclusively on large animal models and this approach has significant limitations in terms of cost and complexity of the studies. Hence, there is a pressing demand for the development of comprehensive, multi-parametric screening strategies that provide improved predictability of cardiotoxic effects.

The central hypotheses of this project are that: (i) human induced pluripotent stem cell (iPSC)-derived cardiomyocyte cultures constitute an effective organotypic culture model for predictive toxicity screening of environmental chemicals; (ii) a population-based experimental design can assess variation in toxicity to better characterize uncertainties; and (iii) integration of pharmacokinetic and high-throughput screening data further improves confidence in NexGen health assessments. We are testing these hypotheses by pursuing the following specific objectives.

Specific Objective 1: To conduct population-based concentration-response high-content/-throughput in vitro screening of up to 200 ToxCast chemicals in human iPSC-derived cardiomyocytes from 100 individuals.

Specific Objective 2: To demonstrate how in vitro testing-derived phenotypes are informative of the molecular and cellular events in the adverse outcome pathway (AOP) for cardiotoxicity.

Specific Objective 3: To collect pharmacokinetic data using human hepatocytes and plasma to perform in vitro-to-in vivo extrapolations of the dose-response information from cardiotoxicity screening to human exposures.

Progress Summary:

Most of the work in the past year has focused on the activities under all proposed Specific Objectives. To date, we have accomplished the following:

  • We have completed testing 140 compounds selected in consultation with FDA, NTP and EPA-NCCT in a population-based in vitro iPSC-derived cell model with iCell cardiomyocytes from 43 normal donors. Several manuscripts detailing the outcomes have been published or are in submission.
  • Using these data from this in vitro screening, and data from human clinical trials, we demonstrated that in vitro human iPSC-derived cardiomyocyte model accurately predicts clinical concentration-QTc relationships. This finding confirms our central hypothesis that human induced pluripotent stem cell (iPSC)-derived cardiomyocyte cultures constitute an effective organotypic culture model for predictive toxicity screening.
  • We also showed that iPSC-derived cardiomyocytes are a highly reproducible in vitro model where donor-specific differences in baseline function and drug-induced effects are highly conserved across experiments and batches of cells. We demonstrated the feasibility of using a panel of population-based organotypic cells from healthy donors as an animal replacement experimental model.
  • The results of in vitro iPSC cell-derived cardiomyocyte assays were corrected for protein binding in cell culture media to enable confident comparisons of dose with in vivo human data. Additional experiments are under way to characterize binding parameters of additional compounds that are tested.
  • We have prepared screening plates with 1000+ chemicals for additional experiments in human iPSC-derived cardiomyocytes from selected donors (5 individuals were selected). These compounds were provided by EPA NCCT and include chemicals of diverse classes with high-throughput toxicokinetic data, established toxicity values and those that are on the TSCA active inventory. The first round of experiments with this library in one iPSC cardiomyocyte cell line was completed. Additional experiments are on the way.

These experiments and observations are significant because they demonstrate novel possibilities of utilizing this microphysiological system for read-across and grouping of the individual chemicals, mixtures, complex substances.

Future Activities:

We will continue experiments in 5 donors with 1000 chemicals.

  • We will collect samples and conduct data analysis for TempO-seq high-throughput transcriptomics on 1000 chemicals in the standard iPSC-cardiomyocyte donor.
  • We will continue collecting protein binding and other kinetic data to enable in vitro-to-in vivo comparisons.
  • We will work closely with Project 3 staff to analyze the data from high-content screening and high-throughput transcriptomics.
  • We will work with project 2 staff to conduct screening of mouse-derived embryoid bodies.


Journal Articles on this Report : 8 Displayed | Download in RIS Format

Other subproject views: All 37 publications 11 publications in selected types All 11 journal articles
Other center views: All 192 publications 35 publications in selected types All 35 journal articles
Type Citation Sub Project Document Sources
Journal Article Blanchette AD, Grimm FA, Dalaijamts C, Hsieh NH, Ferguson K, Luo YS, Anson B, Rusyn I, Chiu WA. Thorough QT/QTc in a dish:An in vitro human model that accurately predicts clinical concentration-QTc relationships. Clinical Pharmacology and Therapeutics 2019;105:1175-1186. R835802 (2018)
R835802 (Final)
R835802C001 (2018)
  • Abstract: Abstract
  • Journal Article Chiu WA, Axelrad DA, Dalaijamts C, Dockins C, Shao K, Shapiro AJ, Paoli G. Beyond the RfD:Broad application of a probabilistic approach to improve chemical dose-response assessments for noncancer effects. Environmental Health Perspective 2018;126(6):067009. R835802 (2018)
    R835802 (Final)
    R835802C001 (2018)
  • Full-text from PubMed
  • Abstract from PubMed
  • Full-text: Environmental Health Perspectives - Full Text HTML
  • Journal Article Fantke P, Aylward L, Bare J, Chiu WA, Dodson R, Dwyer R, Ernstoff A, Howard B, Jantunen M, Jolliet O, Judson R, Kirchhübel N, Li D, Miller A, Paoli G, Price P, Rhomberg L, Shen B, Shin HM, Teeguarden J, Vallero D, Wambaugh J, Wetmore BA, Zaleski R, McKone TE. Advancements in life cycle human exposure and toxicity characterization. Environmetnal Health Perspective 2018;126:125001. R835802 (2018)
    R835802 (Final)
    R835802C001 (2018)
  • Full-text from PubMed
  • Abstract from PubMed
  • Full-text: Environmental Health Perspectives - Full Text HTML and PDF
  • Journal Article Grimm FA, Blanchette A, House JS, Ferguson K, Hsieh NH, Dalaijamts C, Wright AA, Anson B, Wright FA, Chiu WA, Rusyn I. A human population-based organotypic in vitro model for cardiotoxicity screening. ALTEX 2018;35:441-452. R835802 (2018)
    R835802 (Final)
    R835802C001 (2018)
    R835802C003 (2018)
  • Full-text from PubMed
  • Abstract from PubMed
  • Abstract: Abstract
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  • Other: ALTEX - Full Text PDF
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  • Journal Article Grimm FA, House JS, Wilson MR, Sirenko O, Iwata Y, Wright FA, Ball N, Rusyn I. Multi-Dimensional in Vitro Bioactivity Profiling for Grouping of Glycol Ethers. Regulatory Toxicology and Pharmacology 2019;101:91-102. R835802 (2018)
    R835802 (Final)
    R835802C001 (2018)
    R835166 (Final)
  • Abstract from PubMed
  • Full-text: Science Direct - Full Text HTML
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  • Journal Article Konganti K, Ehrlich A, Rusyn I, Threadgill DW. gQTL:a web application for QTL analysis using the collaborative cross mouse genetic reference population. G3:Genes, Genomes, Genetics 2018;8(8):2559-2562 R835802 (2017)
    R835802 (2018)
    R835802 (Final)
    R835802C001 (2018)
    R835802C002 (2018)
  • Abstract from PubMed
  • Full-text: G3-Full Text PDF
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  • Abstract: G3-Abstract
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  • Journal Article Marvel SW, To K, Grimm FA, Wright FA, Rusyn I, Reif DM. ToxPi Graphical User Interface 2.0: dynamic exploration, visualization, and sharing of integrated data models. BMC Bioinformatics 2018;19(1):80 (7 pp.). R835802 (2017)
    R835802 (2018)
    R835802 (Final)
    R835802C001 (2018)
    R835802C003 (2018)
  • Full-text from PubMed
  • Abstract from PubMed
  • Associated PubMed link
  • Full-text: BMC Bioinformatics-Full Text HTML
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  • Abstract: BMC Bioinformatics-Abstract
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  • Other: BMC Bioinformatics-Full Text PDF
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  • Journal Article Hsieh NH, Reisfeld, B, Bois FY, Chiu WA. Applying a global sensitivity analysis workflow to improve the computational efficiencies in physiologically-based pharmacokinetic modeling. Frontiers in Pharmacology 2018 9:588. R835802 (2018)
    R835802 (Final)
    R835802C001 (2018)
  • Full-text: Full Text
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  • Abstract: Abstract
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  • Supplemental Keywords:

    cardiovascular, stem cells, toxicity pathway, variability, pharmacokinetic model.

    Progress and Final Reports:

    Original Abstract
  • 2015 Progress Report
  • 2016
  • 2017

  • Main Center Abstract and Reports:

    R835802    Organotypic Culture Models For Predictive Toxicology Center

    Subprojects under this Center: (EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
    R835802C001 High-throughput Hazard,Dose-responseandPopulationVariabilityAssessmentof Cardiotoxicity in aHumanInducedPluripotentStem Cell(iPSC)-derivedin vitro Culture Model
    R835802C002 Linking in vitro-to-in vivoToxicity Testing Using Genetically-matchedOrganoids and Mice from a Novel Genetic Reference Population
    R835802C003 A Pipeline for in vitro-to-in vivo Extrapolation, Population Modeling, & Prioritization