Organotypic Model of Human Kidney as a Platform for Adverse Outcomes Pathway Assessment of Engineered Nanomaterials

EPA Grant Number: R835738C002
Subproject: this is subproject number 002 , 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 Model of Human Kidney as a Platform for Adverse Outcomes Pathway Assessment of Engineered Nanomaterials
Investigators: Kelly, Edward J.
Institution: University of Washington
EPA Project Officer: Klieforth, Barbara I
Project Period: December 1, 2014 through November 30, 2018
Project Amount: Refer to main center abstract for funding details.
RFA: Organotypic Culture Models for Predictive Toxicology Center (2013) RFA Text |  Recipients Lists
Research Category: Safer Chemicals , Health , Human Health

Objective:

The primary purpose of this project is to design, implement and test a tissue engineered human kidney microphysiological system. The system will be developed to fully evaluate uptake, metabolism and elimination of xenobiotics in a human tissue derived, in vitro 3-dimensional system that accurately reflects human physiology. The microphysiological system can be used to predict disposition kinetics of xenobiotics, and also assess the response to kidney injury inflicted by endogenous and exogenous toxicants, as well as from pathogens.

Approach:

For this project we will focus on the ENMs (QDs and AgNPs) that have practical applications in photonics, optics, solar cells, sensors, imaging, and anti-microbial activity.
Our hypotheses for this project are:
1) Differentiation status of organotypic kidney cultures will define the response to ENMs.
2) The oxidative stress and ER stress responses within organotypic kidney cultures will be key common response pathways associated with exposure to ENMs.
3) Inflammatory pathways within organotypic kidney cultures will have significant and specific impacts beyond general oxidative stress.
4) Genetic differences among organotypic kidney cultures will be critical factors in interpreting kidney specific responses.
5) Using a systems-based Adverse Outcome Pathway (AOP) analysis with toxicokinetic and dynamic models as a part of a risk assessment framework will allow for cross assay and organ interpretation.
In order to address these hypotheses, this project will focus on developing an organotypic kidney model systems derived from human kidney. We will use a microphysiometer flow-through system developed by Nortis Inc, to establish and differentiate long-term cultures of kidney proximal tubule cells for this project.

Rationale:

The establishment of functional differentiation over long-term culture is necessary to demonstrate the suitability of the Nortis microphysiometer device as a model of human kidney.
These human kidney models (primary and iPSCs induced PTECs) will useful for the validation of AOPs caused by exposure to metal-based ENMs, especially since the kidney is often a target of metals. Moreover, the incorporation of human individual susceptibility into this model system address a critical unmet need in human risk associated with ENM exposure. Ultimately, the goal of this Project is to compare these models to that of humans experiencing real world exposures to these ENMs.
 
ENMs continue to expand in scope of use and production, yet knowledge regarding their potential toxicity continues to dramatically lag their development. This represents a real risk not
only to the potential impacts they may have on human and environmental health, but has the potential to stifle progress in this industry if principles governing their toxicity are not discovered and mitigated. Moreover, very little is known regarding their mechanism of action and adverse outcomes associated with exposure to these ENMs. Thus, this Project will address several important needs, including the role of surface and core constituents on uptake, toxicity and disposition of selected metal containing ENMs, and the role of individual susceptibility as a factor influencing toxicity.

 

Expected Results:

• We will establish and standardize advanced organotypic cell culture techniques to model kidney responses to ENMs and other relevant compounds. This comprehensive assessment
will include a cellular model of the normal human kidney with emphasis on determinants of differentiation and response to ENMs.
• Establishment of a human kidney model that has been transduced with specific reporter constructs (Grx1-roGFP and roGFP2-Orp1) will allow mid- to high-throughput screening of
different compounds.
• We will identify the specific markers that are most useful in defining dynamic cellular responses to nanomaterial and other chemical exposures to the kidney. Categories of
responses that will be evaluated include cellular injury as assessed by KIM1 release, LDH, and MTS assay, inflammation as assessed by a range of relevant cytokines, and markers of
cellular stress, including oxidative and ER stress allowing us to predict determinants of differentiation and ENM toxicity.
• New information and methods to make better informed, more timely decisions about chemicals
• Innovative toxicity testing methods and new prediction techniques in computational toxicology that integrate multiple fields of science and technology to better understand the
basis of lifestage- specific susceptibilities

 

Publications and Presentations:

Publications have been submitted on this subproject: View all 23 publications for this subprojectView all 74 publications for this center

Journal Articles:

Journal Articles have been submitted on this subproject: View all 5 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