Grantee Research Project Results
2015 Progress Report: Organotypic Model of Testis as a Platform for Adverse Outcomes Pathway Assessment of Engineered Nanomaterials
EPA Grant Number: R835738C004Subproject: 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: Center for Air, Climate, and Energy Solutions
Center Director: Robinson, Allen
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: Aja, Hayley
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: Chemical Safety for Sustainability
Objective:
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 three-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.
Progress Summary:
In order to fulfill our objective of incorporating developmental and genetic susceptibility into metal and ENM evaluation, we have constructed a detailed timeline relating testicular development and hormonal changes across rats, mice, and humans. This substantiates our recent publication, Wegner, et al., 2015, which was featured on the cover of Reproductive Toxicology. In this publication, we conducted a quantitative pathway analysis of gene expression over time to summarize key pathways that occur during development. This approach has provided a framework for identifying and quantifying the perturbations of normal developmental signaling dynamics caused by environmental factors.
Our previous research has shown that these systems can distinguish between developmentally toxic and developmentally nontoxic phthalates by assessing changes in transcriptomic profiles (Yu, et al., 2009). This year, we have completed the evaluation of more than 80 chemicals for reproductive and developmental toxicity and are using the results from our co-culture system to determine the value of specific endpoints, cytotoxicity, changes in cytokine expression, and testosterone perturbation for the predictive profiles for these agents. The cytotoxicity results were compared to results from the ToxCast and ToxRef databases to identify known reproductive and developmental toxicants. The Benchmark Concentration (BMC) that produced cytotoxicity in 10% of the samples (BMC10) was calculated using EPA’s Benchmark Dose modeling approach.
We also have demonstrated that the testis co-culture system can mount biologically relevant responses following chemical exposure. For example, we have shown that the system can identify chemicals that perturb testosterone production in a dose-dependent manner. We measured testosterone production in our co-culture system at 24 and 72 hours. As the dosages of sodium arsenite, crizotinib, nicotine, valproic acid, and vinclozolin were increased, testosterone production was perturbed. For these chemicals, cytotoxicity and testosterone perturbation occurred at some overlapping doses. Vinclozalin, a fungicide, significantly modified testosterone production at concentrations that were not cytotoxic.
The co-culture system can successfully generate a proinflammatory cytokine response following exposure to phthalates and other compounds (data not shown). Increases in proinflammatory cytokines were observed at 2, 8, and 24 hours following phthalate exposure. Another key focus for this year, related to immune responses, has been the detection of resident macrophages in the testis co-culture (data not shown). These previous results have provided a strong foundation of applications for assessment of lifestage specific reproductive and developmental toxicity of additional metal and ENM compounds.
Future Activities:
Our progress to date has focused on expanding the applications of our testis co-culture system to answer this project’s research questions. The next steps with this work will expand the chemicals tested to include more metals and ENMs. As more biological responses are observed, such as oxidative stress and inflammation, we will be able to better characterize lifestage specific AOPs for reproductive and developmental toxicity.
Journal Articles on this Report : 3 Displayed | Download in RIS Format
Other subproject views: | All 43 publications | 13 publications in selected types | All 13 journal articles |
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Other center views: | All 150 publications | 50 publications in selected types | All 49 journal articles |
Type | Citation | ||
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Harris S, Hermsen SA, Yu X, Hong SW, Faustman EM. Comparison of toxicogenomic responses to phthalate ester exposure in an organotypic testis co-culture model and responses observed in vivo. Reproductive Toxicology 2015;58:149-159. |
R835738 (2016) R835738 (2017) R835738C004 (2015) R835738C004 (2017) R834514 (Final) R834514C003 (2015) R834514C003 (Final) |
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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. |
R835738 (2016) R835738 (2017) R835738C004 (2015) R835738C004 (2016) R835738C004 (2017) R833772 (2009) R834514 (Final) R834514C003 (Final) |
Exit Exit Exit |
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Wegner SH, Yu X, Pacheco Shubin S, Griffith WC, Faustman EM. Stage-specific signaling pathways during murine testis development and spermatogenesis: a pathway-based analysis to quantify developmental dynamics. Reproductive Toxicology 2015;51:31-39. |
R835738 (2016) R835738 (2017) R835738C004 (2015) R835738C004 (2017) R834514 (2015) R834514 (Final) R834514C003 (2015) R834514C003 (Final) |
Exit Exit Exit |
Supplemental Keywords:
reproductive and developmental toxicity, chemical screening, testicular development, in vitro modelRelevant Websites:
The Predictive Toxicology Center (PTC) for Organotypic Cultures ExitProgress and Final Reports:
Original AbstractMain Center Abstract and Reports:
R835738 Center for Air, Climate, and Energy Solutions 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
The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.
Project Research Results
13 journal articles for this subproject
Main Center: R835738
150 publications for this center
49 journal articles for this center