Airway Epithelium Organotypic Culture as a Platform for Adverse Outcomes Pathway Assessment of Engineered Nanomaterials

EPA Grant Number: R835738C001
Subproject: this is subproject number 001 , 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: Airway Epithelium Organotypic Culture as a Platform for Adverse Outcomes Pathway Assessment of Engineered Nanomaterials
Investigators: Altemeier, William
Institution: University of Washington
EPA Project Officer: Klieforth, Barbara I
Project Period: December 1, 2014 through November 30, 2018
RFA: Organotypic Culture Models for Predictive Toxicology Center (2013) RFA Text |  Recipients Lists
Research Category: Safer Chemicals , Health , Human Health

Objective:

The overall goal of this proposal is to utilize organotypic culture systems to better evaluate the potential for cellular and organ toxicity within an adverse outcomes pathway (AOP) model. In this project, we will focus on a model system representing the airway epithelium. The lungs are a major route of exposure to environmental compounds and microbes, which are inhaled during respiration. Clearance of inhaled materials can be accomplished both by mucociliary transport of the airway epithelium followed by expectoration and by phagocytosis by resident macrophages. However, environmental exposures can also cause cellular injury, chronic inflammation and alterations in cellular differentiation, leading to chronic lung injury. Evaluation of the cellular effects of different compounds is an essential first step in assessing the health risks associated with exposure. Traditionally, these experiments have been performed using transformed cell lines, which allow easy scalability to higher throughput analyses. However, there has been growing recognition that transformed cell lines do not recapitulate responses observed in primary cells, particularly when these cells are differentiated to a phenotype resembling their in vivo state.

In this project, we will utilize primary tracheal epithelial cell culture that has been differentiated at an air-liquid interface (ALI), representing a three-dimensional organotypic culture. Murine tracheal epithelial culture (mTEC) will be used initially as this allows use of cell isolation from genetically modified animals both for pathway analysis and for development of medium throughput assays. Importantly, mTEC organotypic culture can be differentiated at ALI towards a “normal” phenotype with more ciliated epithelium and fewer mucus producing cells. Alternatively, differentiation at ALI in the presence of IL-13 skews epithelial differentiation towards a “disease phenotype” characterized by an increase in mucus producing cells. Additionally, mTEC ALI contains Club cells (aka Clara cells), which are important progenitor cells required for epithelial repair following injury and pleuripotent basal cells from which other cells in the system are derived. Thus, mTEC ALI culture represents a true organotypic mixed cellular model of the airways. Finally, commercially available human tracheal airway cell culture (hTEC) can be acquired and differentiated under ALI conditions. These will be used to validate specific findings identified in mTEC-ALI cultures.

Publications and Presentations:

Publications have been submitted on this subproject: View all 5 publications for this subprojectView all 74 publications for this center

Journal Articles:

Journal Articles have been submitted on this subproject: View all 1 journal articles for this subprojectView all 13 journal articles for this center

Progress and Final Reports:

2015 Progress Report
2016 Progress Report


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 for Adverse Outcomes Pathway 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