Organotypic Liver Model for Predictive Human Toxicology and Metabolism

EPA Grant Number: R835736C004
Subproject: this is subproject number 004 , 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: Organotypic Liver Model for Predictive Human Toxicology and Metabolism
Investigators: Taylor, D. Lansing , Davidson, Jeffrey M. , Gough, Albert , Vernetti, Lawrence
Institution: University of Pittsburgh - Main Campus , 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:

(1) Transfer current microfluidics-based, 4-cell type, 3D liver organoid into commercial device; (2) demonstrate normal and abnormal liver function, drug metabolism and toxicity responses for 50-60 ToxCast chemicals; (3) demonstrate IM-MS analysis of targeted secretome products including drug metabolites, cytokines, lipids, proteins; (4) implement realtime, fluorescence-based biosensors of key physiological functions for liver and other organoid culture models (OCMs); (5) provide liver+drug conditioned media to other OCMs; (6) validate iPSC hepatocytes for use in the device; (7) adapt University of Pittsburgh Microphysiology Database to manage, consolidate, visualize, QC and organize data for transfer to EPA. 

Approach:

The liver OCM is a reconstruction of a human liver sinusoidal unit amenable for pharmacokinetics, toxicology and pharmacology studies and has demonstrated normal liver functions including drug metabolism up to 3 weeks in culture. A key feature of the liver OCM is the inclusion of hepatocytes, endothelial cells, macrophages and stellate cells – all genetically encoded with fluorescence-based biosensors for High Content Imaging of key pathways and mechanisms of toxicity (oxidative stress, mitochondrial dysfunction, apoptosis, immune and fibrosis response). The liver organoid will be integrated into Vanderbilt’s moderate throughput screening platform as a microfluidic device to allow use of the secreted media for: (a) assaying the secretome in near real-time for targeted high content metabolomics; (b) provide liver conditioned media to feed chemical metabolites to the reproductive and developmental OCMs, and (c); track mechanisms of toxicity in the liver. During the project we propose to replace the primary hepatocytes used in our current device with renewable, validated, adult-like iPSC hepatocytes and develop novel biosensors to AhR, ppar-a, NFkB, PR and others for incorporation into the 4 OCMs. Finally, we will adapt the UPDDI Microphysiology database to manage data, conduct analysis, QC and transfer the data to the EPA. 

Expected Results:

The outputs of this project will include: (1) a metabolically active liver OCM linked to other OCMs with the capacity to be used for predictive Adverse Outcome Pathway elucidation on a moderate throughput screening platform; (2) presentations and publications that advance the understanding how liver pre-treatment of chemicals can lead to human developmental abnormalities; (3) identification of liver-related mechanism(s) which interfere with normal reproductive and development function. 

Publications and Presentations:

Publications have been submitted on this subproject: View all 3 publications for this subprojectView all 64 publications for this center

Journal Articles:

Journal Articles have been submitted on this subproject: View all 3 journal articles for this subprojectView all 15 journal articles for this center

Supplemental Keywords:

biology, cellular, human, human health, molecular, systems based model, teratogen, chemicals, toxins, hepatotoxins, development, risk assessment, methods

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