2016 Progress Report: Organotypic Model of Testis as a Platform for Adverse Outcomes Pathway Assessment of Engineered NanomaterialsEPA Grant Number: R835738C004
Subproject: this is subproject number 004 , 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 Testis as a Platform for Adverse Outcomes Pathway Assessment of Engineered Nanomaterials
Investigators: Faustman, Elaine
Institution: University of Washington
EPA Project Officer: Klieforth, Barbara I
Project Period: December 1, 2014 through November 30, 2018
Project Period Covered by this Report: December 1, 2015 through November 30,2016
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
The overall goal of this project is to utilize an organotypic in vitro model of testicular development to evaluate the male reproductive toxicity of engineered nanomaterials (ENM) using an adverse outcome pathway (AOP) framework. We will use 3-dimensional in vitro testicular co-cultures that have been shown to capture key processes of male reproductive development in order to evaluate the potential for ENMs to alter these processes. We will measure the ability of ENMs to alter cellular differentiation and tissue maturation with a focus on the roles of developmental timing and genetics in influencing susceptibility. We also will explore the role of oxidative stress and inflammation pathways in mediating ENM induced perturbation of development. Finally, we will use toxicokinetic and dynamic models to integrate in vitro our findings into an AOP framework.
Establishing a life stage context for our testis organotypic cultires:
We have measured testosterone production and protein expression in our co-culture system at days in vitro (DIV) 2, 3, 6, 7, 15, and 16 for baseline characterization. We then compared our results to transcriptomes from publicly available in vivo mouse and rat testes and our in vitro rat testes data.
For evaluation of cadmium’s developmental toxicity, we have measured cytotoxicity, cell viability, and morphology at 24 hours after treatment at different developmental stages (DIV 2, 6, and 15). We have observed a dose and life stage-dependent disruption in these measures. Significant dose-related decline in testosterone production was observed for DIV 3, while little change was observed for DIV 7 and DIV 16. Significant dose response was observed for LDH for all three time points, while DIV 16 has the most dramatic effects. Our results demonstrate the potential of our co-culture systems and system biology model to capture adverse outcomes of male reproductive development.
Our progress to date has focused on: (1) expanding the applications of our testis co-culture system to answer this project’s research questions through development of organotypic cultures to evaluate reproductive and developmental toxicity, (2) modification of our previous co-culture system for immature mouse testes, (3) characterization of normal testes development, and (4) evaluation of perturbed conditions using various chemicals including cadmium. We plan to finalize our characterization of life-stage specific AOPs for reproductive and developmental toxicity (papers in progress). We also plan to evaluate the role of genes in response to AgNP by utilizing our co-culture systems from two strains of mice (AJ and C57BL/6). We will finalize our assessment of the toxicological effects and benchmark dose of various silver nanoparticles on A/J mouse midbrain micromass, building upon our findings for the C57BL/6 mouse midbrain micromass experiment.
Journal Articles on this Report : 2 Displayed | Download in RIS Format
|Other subproject views:||All 35 publications||6 publications in selected types||All 6 journal articles|
|Other center views:||All 134 publications||39 publications in selected types||All 38 journal articles|
||Harris S, Wegner S, Hong SW, Faustman EM. Phthalate metabolism and kinetics in an in vitro model of testis development. Toxicology in Vitro 2016;32:123-131.||
||Harris S, Shubin SP, Wegner S, Van Ness K, Green F, Hong SW, Faustman EM. The presence of macrophages and inflammatory responses in an in vitro testicular co-culture model of male reproductive development enhance relevance to in vivo conditions. Toxicology In Vitro 2016;36:210-215.||
Supplemental Keywords:reproductive and developmental toxicity, chemical screening, testicular development, in vitro model
Relevant Websites:The Predictive Toxicology Center (PTC) for Organotypic Cultures Exit
Progress and Final Reports:Original Abstract
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