Mammosphere Bioreactor For Life-Stage Specific Toxicology

EPA Grant Number: R835736C001
Subproject: this is subproject number 001 , established and managed by the Center Director under grant R835736
(EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).

Center: Vanderbilt–Pittsburgh Resource for Organotypic Models for Predictive Toxicology (VPROMPT)
Center Director: Hutson, Michael Shane
Title: Mammosphere Bioreactor For Life-Stage Specific Toxicology
Investigators: McCawley, Lisa J. , Markov, Dmitry
Current Investigators: McCawley, Lisa J. , Auner, Alex , Bazilevich, Gloria , Fryman, Phillip , Markov, Dmitry , McIntyre, J. Oliver , Schafer, Emily
Institution: University of Pittsburgh - Main Campus , Vanderbilt University
Current Institution: Vanderbilt University
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:

Taking advantage of unique collaborations afforded through VPROMPT, we will develop a microfluidic Mammary Gland organotypic culture model (MG-OCM) for toxicant assessment, for monitoring dynamic toxicant-dependent changes to cellular functions and signaling cascades, and elucidation of Adverse Outcome Pathways. Specific objectives are: (A) to develop and validate the predictive utility of self-contained, fully-automated MG-OCM modules; (B) to investigate the effects of chemical exposures on MG-OCMs for chemicals shown to reduce lactation index in only the F2 generation of multigenerational studies – strongly suggesting a role for life-stage specific exposure; and (C) to develop and validate toxicant assessment for compounds requiring metabolic activation using a paired liver-OCM/MG-OCM. 

Approach:

We will extend our previously developed, microfabricated Mammary Gland Thick Tissue Bioreactor (MG-TTB) to support the combination of 3D culture methods with controlled exposure to investigate chemicals for potential mammary toxicity. The TTB system offers significant advantages over traditional 3D cell culture, allowing for decreased reagent use and physiologically relevant microenvironments. We will evaluate a tri-culture system (normal human mammary epithelial cells + human mammary fibroblasts + sub-cutaneous adipocytes) for a more accurate recapitulation of mammary gland biology and crosstalk between heterotypic cells. We will exploit this system for in vitro evaluation of potential environmental toxicants for effects on mammary development using both chronic and acute exposures at various stages of gland development. Furthermore, this system will be used for high content analysis of putative toxicants with a focus on key biomarkers: matrix proteinase activity, molecular signaling cascades, metabolic alterations, and changes in cytokine and growth factor expression. We will also conduct unbiased evaluation of cellular responses by analyzing the secretome present in MG-OCM effluent using Ion-Mobility Mass Spectrophotometry. 

Expected Results:

  1. Functional in vitro MG-OCMs that closely mimic and predict in vivo responses to acute and systemic chemical exposures.
  2. A set of self-contained and automated MG-OCMs with sequential improvements toward medium throughput and/or high content screening of chemical exposures to assess life-stage specific toxicity.
  3. A paired liver-OCM/MG-OCM platform to study the effects of toxicant metabolites and/or abnormal liver behavior on MG development.

Publications and Presentations:

Publications have been submitted on this subproject: View all 5 publications for this subprojectView all 64 publications for this center

Supplemental Keywords:

biology, exposure, risk assessment, dose-response metabolism, toxins, human health, innovative technology, systems biology

Progress and Final Reports:

2015 Progress Report
2016 Progress Report


Main Center Abstract and Reports:

R835736    Vanderbilt–Pittsburgh Resource for Organotypic Models for Predictive Toxicology (VPROMPT)

Subprojects under this Center: (EPA does not fund or establish subprojects; EPA awards and manages the overall grant for this center).
R835736C001 Mammosphere Bioreactor For Life-Stage Specific Toxicology
R835736C002 Organotypic Culture Model to Analyze Developmental Limb Malformations Resulting from Toxicant/Teratogen Exposure
R835736C003 Validating a fetal membrane on a chip model for characterizing reproductive toxicant exposure risks
R835736C004 Organotypic Liver Model for Predictive Human Toxicology and Metabolism
R835736C005 Systems Engineering & Analysis for Organotypic Culture Models