2015 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 , North Carolina State University , University of North Carolina at Chapel Hill
Current 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, 2015 through May 31,2016
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 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 proposed Specific Objective 1. To date, we have accomplished the following:

  • We developed optimal assay multi-plexing for high-content imaging and high-throughput transcriptomic analyses using iCell cardiomyocyte organotypic culture model and other cell types derived from iPS cells. Overall, our results demonstrate how a compendium of assays can be utilized for quantitative screening of chemical effects in iPSC cardiomyocytes and hepatocytes and enable rapid and cost-efficient multidimensional biological profiling of toxicity. These experiments were detailed in Grimm, et al. (2015). 
  • By using a compendium of high-content screening assays and several (iPSC)-derived cells cell types we have conducted toxicological categorization of eight propylene (P-series) and twelve ethylene (E-series) glycol ethers, structurally related yet toxicologically diverse group of prototypical industrial high production volume chemicals. These data were used to conduct grouping of substances using biological profiling. We found that there is a correlation between the length of the alcohol group and induced effects such that glycol ethers can be categorized based on simple glycols, methyl-, ethyl-, propyl-, butyl-, and hexyl ethers. These data were presented at 2016 SOT meeting (Iwata, et al., 2016).

  • We also conducted experiments in iPSC-derived cardiomyocytes and hepatocytes with extracts of 26 petroleum substances comprising six product groups. The data revealed group-specific clustering and a high degree of correlation between biological and chemical data sets. Altogether, we demonstrate how novel analytical chemistry and in vitro screening approaches can be effectively utilized to categorize UVCBs thereby indicating their potential applicability in regulatory submissions. These experiments were detailed in Grimm, et al. (2016) publication.

  • We have finalized the list of 140 compounds to be used in screening by Projects 1 and 2. Chemical selection was coordinated with FDA, NTP and EPA-NCCT. All compounds have been procured, dilutions prepared, and chemical master plates created for screening.

  • Screening of these 140 chemicals in a population-based in vitro iPSC-derived cell model commenced with iCell cardiomyocytes from 30 normal donors (European and African American descent, equal male to female ratio).

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

Future Activities:

  • We will perform data analyses on the first batch of 30 individuals screened against 140 chemicals and appropriate positive and negative controls.
  • We expect to receive the next batch of cells (15-20 donors) from CDI in October-November 2016 and additional batches of 10-15 donors at quarterly intervals thereafter. These cells will be screened and data analyzed.
  • We plan to conduct several large batches of targeted transcriptomic analyses using S1500+ gene list (~2,700 targets) developed by the National Toxicology Program for TempO-seq high-throughput transcriptomics.
  • 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. Towards the end of year 2 and into subsequent years we will begin conducting experiments for reverse toxicokinetics based on the data for 140 chemicals that are screened. 


Journal Articles on this Report : 3 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 Cote I, Andersen ME, Ankley GT, Barone S, Birnbaum LS, Boekelheide K, Bois FY, Burgoon LD, Chiu WA, Crawford-Brown D, Crofton KM, DeVito M, Devlin RB, Edwards SW, Guyton KZ, Hattis D, Judson RS, Knight D, Krewski D, Lambert J, Maull EA, Mendrick D, Paoli GM, Patel CJ, Perkins EJ, Poje G, Portier CJ, Rusyn I, Schulte PA, Simeonov A, Smith MT, Thayer KA, Thomas RS, Thomas R, Tice RR, Vandenberg JJ, Villeneuve DL, Wesselkamper S, Whelan M, Whittaker C, White R, Xia M, Yauk C, Zeise L, Zhao J, DeWoskin RS. The next generation of risk assessment multi-year study--highlights of findings, applications to risk assessment, and future directions. Environmental Health Perspectives 2016;124(11):1671-1682. R835802 (2015)
R835802 (2016)
R835802 (2017)
R835802 (2018)
R835802C001 (2015)
R835166 (Final)
  • Full-text from PubMed
  • Abstract from PubMed
  • Associated PubMed link
  • Full-text: EHP-Full Text PDF
  • Abstract: EHP-Abstract and Full Text HTML
  • Journal Article Grimm FA, Iwata Y, Sirenko O, Bittner M, Rusyn I. High-content assay multiplexing for toxicity screening in induced pluripotent stem cell-derived cardiomyocytes and hepatocytes. Assay and Drug Development Technologies 2015;13(9):529-546. R835802 (2015)
    R835802 (2016)
    R835802 (2017)
    R835802 (2018)
    R835802C001 (2015)
  • Full-text from PubMed
  • Abstract from PubMed
  • Associated PubMed link
  • Full-text: Mary Ann Liebert-Full Text HTML
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  • Abstract: Mary Ann Liebert-Abstract & Full Text HTML
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  • Other: Mary Ann Liebert-Full Text PDF
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  • Journal Article Grimm FA, Iwata Y, Sirenko O, Chappell GA, Wright FA, Reif DM, Braisted J, Gerhold DL, Yeakley JM, Shepard P, Seligmann B, Roy T, Boogaard PJ, Ketelslegers HB, Rohde AM, Rusyn I. A chemical-biological similarity-based grouping of complex substances as a prototype approach for evaluating chemical alternatives. Green Chemistry 2016;18(16):4407-4419. R835802 (2015)
    R835802 (2016)
    R835802 (2017)
    R835802 (2018)
    R835802C001 (2015)
    R835166 (Final)
  • Full-text from PubMed
  • Abstract from PubMed
  • Full-text: Royal Society of Chemistry-Full Text PDF
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  • Abstract: Royal Society of Chemistry-Abstract
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  • Other: ResearchGate-Abstract & Full Text PDF
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  • Supplemental Keywords:

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

    Progress and Final Reports:

    Original Abstract
  • 2016
  • 2017
  • 2018 Progress Report

  • 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