Grantee Research Project Results
Final Report: Instrumenting phenotypic immunological responses to toxicants that threaten human reproduction
EPA Grant Number: R839501Title: Instrumenting phenotypic immunological responses to toxicants that threaten human reproduction
Investigators: Osteen, Kevin G , Cliffel, David , McLean, John , Bruner-Tran, Kaylon L.
Institution: Vanderbilt University Medical Center , Vanderbilt University
EPA Project Officer: Callan, Richard
Project Period: August 1, 2019 through July 31, 2022 (Extended to July 31, 2024)
Project Amount: $848,923
RFA: Advancing Actionable Alternatives to Vertebrate Animal Testing for Chemical Safety Assessment (2018) RFA Text | Recipients Lists
Research Category: Chemical Safety for Sustainability
Objective:
The primary goals of our project were to 1) examine whether a pre-existing inflammatory condition impacts the endometrium’s acute/chronic responses to an environmental toxicant known to impact fertility and 2) develop microfluidic in vitro models that can utilize primary human reproductive tract cells and/or cell lines that can serve as appropriate alternatives to animal studies for accurate endpoint assessments.
Summary/Accomplishments (Outputs/Outcomes):
The human endometrium undergoes a monthly, development-like regeneration process; therefore, cellular differentiation in preparation for pregnancy is at an environmental risk during each menstrual cycle. Endometriosis, an inflammatory reproductive tract disease of women, is associated with reduced endometrial progesterone (P4) responsiveness. Consequently, these women are at risk of infertility/subfertility and, if they become pregnant, a heightened risk of adverse pregnancy outcomes. The endometrial “endometriosis phenotype” is further marked by a reduced ability of P4 to suppress matrix metalloproteinases (MMPs), enzymes which regulate matrix turnover. Significantly, a similar endometriosis phenotype was observed in mice with a direct developmental exposure to TCDD as well as in three additional generations. To translate our animal models to women, our EPA-funded studies utilized our organ-on-a-chip model of the menstrual cycle (EndoChip) to determine how an individual’s preexisting inflammatory disease alters their sensitivity to toxicant exposure.
Conclusions:
Our EPA funded research included a number of significant observations as well as the development or use of new methodology, including:
-Incorporated the use of endometrial organoids for propagation of primary epithelial cells in a matrix, enabling the expansion of these cells without loss of specialized phenotype.
-Demonstrated a synergistic effect of either disease (endometriosis) or TCDD exposure and infection-mediated immune signaling in the loss of progesterone action.
-Determined that perfusion of TCDD-exposed human immune cells (from control donors) within the EndoChip led to reduced stromal cell progesterone response and impaired decidualization.
-Developed the use of specific biosensors (eg, MMP3) to monitor cellular responses within a microfluidic device.
-Demonstrated Raman Spectroscopy methods can be used to distinguish the biochemical footprint of cells (endometrial and immune) from women with and without endometriosis. Our studies lay the groundwork for the potential use of this technology for bench-to-bedside clinical diagnostics.
- Since cell lines are accessible to most researchers, we initiated Case Studies using endometriosis epithelial (12z) and stromal (22b) cell lines to demonstrate compatibility within the EndoChip.
-Identified TCDD-mediated alterations in endometrial stromal cell expression of the DNA methyltransferase (DNMT) family, enzymes that have a central role in epigenetic gene regulation of progesterone receptors in these cells. Expression of the canonical DNMT enzymes - DNMT1, DNMT3A and DNMT3B were all found to be disrupted by TCDD exposure.
-Developed a modified version of the EndoChip using a human trophoblast cell line in order to create an in vitro model of the maternal-fetal interface of early pregnancy (the EndoChip-MFI).
Based on our EPA studies, and separately funded studies utilizing our mouse model, we currently hypothesize that TCDD exposure at critical timepoints leads to altered immune cell training and contributes to the development of endometriosis and its sequelae.
Journal Articles on this Report : 2 Displayed | Download in RIS Format
| Other project views: | All 23 publications | 5 publications in selected types | All 5 journal articles |
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Stephens VR, Moore RE, Spicer SK, Talbert JA, Lu J, Chinni R, Chambers SA, Townsend SD, Manning SD, Rogers LM, Aronoff DM. Environmental Toxicant Exposure Paralyzes Human Placental Macrophage Responses to Microbial Threat. ACS Infectious Diseases 2023;9(12):2401-8. |
R839501 (Final) |
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Virgous C, Lyons L, Sakwe A, Nayyar T, Goodwin S, Hildreth J, Osteen K, Bruner-Tran K, Alawode O, Bourne P, Hills ER. Resumption of Spermatogenesis and Fertility Post Withdrawal of Hydroxyurea Treatment. International Journal of Molecular Sciences 2023;24(11):9374. |
R839501 (Final) |
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Progress and Final Reports:
Original AbstractThe 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
- 2023 Progress Report
- 2022 Progress Report
- 2021 Progress Report
- 2020 Progress Report
- Original Abstract
5 journal articles for this project