2015 Progress Report: Organotypic Models of Mammalian Liver as a Platform for Adverse Outcomes Pathway Assessment of Engineered Nanomaterials

EPA Grant Number: R835738C003
Subproject: this is subproject number 003 , established and managed by the Center Director under grant R835738
(EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).

Center: Predictive Toxicology Center for Organotypic Cultures and Assessment of AOPs for Engineered Nanomaterials
Center Director: Faustman, Elaine
Title: Organotypic Models of Mammalian Liver as a Platform for Adverse Outcomes Pathway Assessment of Engineered Nanomaterials
Investigators: Kavanagh, Terrance J , Eaton, David
Institution: University of Washington
EPA Project Officer: Klieforth, Barbara I
Project Period: December 1, 2014 through November 30, 2018 (Extended to November 30, 2019)
Project Period Covered by this Report: December 1, 2014 through November 30,2015
RFA: Organotypic Culture Models for Predictive Toxicology Center (2013) RFA Text |  Recipients Lists
Research Category: Safer Chemicals , Health , Human Health

Objective:

To develop an organotypic 3D model of human and rodent liver using a microphysiological device, and evaluate its suitability for assessing the adverse effects of engineered nanomaterials and heavy metals. 

Progress Summary:

Primary human, rat or mouse hepatocytes were used to populate a dual-chamber Nortis microphysiological system. These devices then were evaluated for cell morphology, viability, and functionality (cytochrome P450 activity; HNF4a expression; albumin production). They also were evaluated for their ability to metabolize vitamin D and to metabolize and activate the nephrotoxin aristolochic acid (AA) into metabolites that were toxic toward proximal tubule epithelial cells (PTEC) cultured in tandem in another Nortis MPS device.

Preliminary results indicate that hepatocytes cultured in the 3D MPS maintain greater viability and hepatic functions for at least 14 days, whereas 2D cultures only exhibited these for 5-6 days. Compared to 2D monolayers, hepatocytes in 3D organotypic cultures also exhibit enhanced albumin production, HNF4a expression, cytochrome p450 activity and inducibility (Cyp1A1; Cyp3A4), presence of bile canaliculi, multi-drug resistance protein-2 (MRP2) transporter expression and function, vitamin D hydroxylation, and AA bioactivation (as indicated by a 4- to 10-fold increase in toxicity of AA toward PTEC when AA is passed through the liver prior to PTEC, vs. direct exposure of PTECs to AA). 

Future Activities:

In the near future, we will continue to characterize hepatocyte function in the 3D microphysiological (MPS) system and assess their polarized phenotype (presence of bile canaliculi; xenobiotic transporter expression and localization).  We then will evaluate the effects of silver nanoparticles (AgNPs), quantum dot nanoparticles, silver and cadmium ions, and other heavy metals in hepatocytes cultured in 2D monolayers vs. 3D MPS. These will include quantitative measures of viability, function, the induction of glutathione pathway genes and proteins, metallothionein (MT) expression, and oxidative stress biomarker expression.  In a manner similar to that we have described for tandem organ on a chip evaluation of AA toxicity, we will examine the ability of hepatocyte expression of MT to deliver Cd and Ag to PTECS and determine if such delivery influences kidney cell viability and function. We also have developed lentiviral vectors, which report on cellular redox status, and these will be applied to this system as well as other systems in the UW Predictive Toxicology Center. 

Journal Articles:

No journal articles submitted with this report: View all 20 publications for this subproject

Supplemental Keywords:

3D organotypic cultures, microphysiological systems, hepatocytes, mouse, human, rat, nanoparticles, quantum dots, aristolochic acid, cadmium, silver, cytotoxicity, redox status, cellular stress response. 

Relevant Websites:

The Predictive Toxicology Center (PTC) for Organotypic Cultures Exit

Progress and Final Reports:

Original Abstract
  • 2016 Progress Report
  • 2017 Progress Report
  • 2018 Progress Report
  • Final

  • Main Center Abstract and Reports:

    R835738    Predictive Toxicology Center for Organotypic Cultures and Assessment of AOPs for Engineered Nanomaterials

    Subprojects under this Center: (EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
    R835738C001 Airway Epithelium Organotypic Culture as a Platform forAdverseOutcomesPathway Assessment of Engineered Nanomaterials
    R835738C002 Organotypic Model of Human Kidney as a Platform for Adverse Outcomes Pathway Assessment of Engineered Nanomaterials
    R835738C003 Organotypic Models of Mammalian Liver as a Platform for Adverse Outcomes Pathway Assessment of Engineered Nanomaterials
    R835738C004 Organotypic Model of Testis as a Platform for Adverse Outcomes Pathway Assessment of Engineered Nanomaterials
    R835738C005 Integrating Liver, Kidney and Testis Nanomaterial Toxicity using the Adverse Outcome Pathway Approach