2015 Progress Report: 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
Center Director: Hutson, Michael Shane
Title: Organotypic Liver Model for Predictive Human Toxicology and Metabolism
Investigators: Taylor, D. Lansing , Gough, Albert , Vernetti, Lawrence
Current Investigators: Taylor, D. Lansing , Davidson, Jeffrey M. , Gough, Albert , Vernetti, Lawrence
Institution: University of Pittsburgh Main Campus
Current Institution: University of Pittsburgh Main Campus , Vanderbilt University
EPA Project Officer: Klieforth, Barbara I
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: Safer Chemicals , Health , Human Health

Objective:

The first year objectives of the human liver organotypic culture model (OCM) as part of the VPROMPT Center were multi-faceted. The goal was to transfer our microfluidics based, 4-cell type, 3D liver organoid into a commercial microfluidic device. We also aimed to demonstrate the ability to monitor normal and abnormal liver function by including indicators of cell health such as bile efflux, albumin synthesis, and urea synthesis. In regards to drug metabolism, we wanted to demonstrate that the liver OCM is a fully capable elimination organ, while also separately aiming to identify mechanisms of toxicity (MOTs) for control toxins using functional biosensors. Another milestone concentrated on providing liver-conditioned media to other Project OCMs. 

In addition, we needed to adapt the University of Pittsburgh Microphysiology Database (UPMDb) to manage, consolidate, visualize, QC and organize data for transfer to EPA, while compiling pre-clinical, clinical and epidemiology findings on VPROMPT-relevant chemicals into UPMDb. Finally, we had to make available our current fluorescence-based biosensors of key physiological functions to the other OCM project teams and to develop new biosensors.

Progress Summary:

A liver organotypic culture model (OCM) has been developed and characterized in the Drug Discovery Institute laboratory of D. Lansing Taylor, University of Pittsburgh. The liver OCM has been successfully transferred to a commercial microfluidic device manufactured by Nortis (Seattle, WA) and consists of four human liver cell types that are sequentially layered, then allowed to self-assemble (SQL-SAL) into a 3D tissue-like liver structure.

We have demonstrated that drugs, drug metabolites, albumin, urea, and lactate dehydrogenase (LDH) can be monitored in the efflux media.

In addition, we have shown functional readouts in the liver OCM,  by transducing 20% of primary hepatocytes and/or stellate cells with fluorescent protein biosensors (transduced cells are referred to as sentinel cells), which are used for live readout of apoptosis, reactive oxygen species (ROS), cell organization and cell proliferation by high content analysis (HCA). Further, the fluorescent biosensors have been provided to the other project teams for incorporation into additional OCMs. 

We have provided liver conditioned and cell free media for Ion-Mobility Mass Spectrometry IM-MS secretome analysis, which was used for the putative identification of 106 small organic molecules consumed and 116 small organic metabolites secreted by the liver OCM. In a proof-of-principle demonstration, we used the liver OCM to activate cyclophosphamide to the form capable of killing MDA-MB 231 breast cancer cells. 

Finally, we have added functionality to the MPS-Db to manage data from the VPROMPT Center to 1) upload raw and processed data from the OCMs, 2) support the review and QC of those data, and 3) correlate the data with human and animal compound exposure data with a goal to support the validation of the OCMs and the production of predictive models.

 

Table 1. Example of human organ experimental results and selected pre-clinical, clinical and PK results found in the University of Pittsburgh’s Microphysiology Systems Database. 

Future Activities:

In Year 2, we will test 10-20 ToxCast chemicals on the standalone Liver OCM, continue the IM-MS analysis to compare drug treated and vehicle secretome responses, and start the integration of the Liver OCM onto the Vanderbilt platform for testing. 


Journal Articles on this Report : 2 Displayed | Download in RIS Format

Other subproject views: All 34 publications 10 publications in selected types All 10 journal articles
Other center views: All 149 publications 39 publications in selected types All 39 journal articles
Type Citation Sub Project Document Sources
Journal Article Senutovitch N, Vernetti L, Boltz R, DeBiasio R, Gough A, Taylor DL. Fluorescent protein biosensors applied to microphysiological systems. Experimental Biology and Medicine 2015;240(6):795-808. R835736 (2015)
R835736 (2016)
R835736 (2018)
R835736C004 (2015)
R835736C004 (2016)
  • Full-text from PubMed
  • Abstract from PubMed
  • Associated PubMed link
  • Abstract: Sage Publications-Abstract
    Exit
  • Journal Article Vernetti LA, Senutovitch N, Boltz R, DeBiasio R, Shun TY, Gough A, Taylor DL. A human liver microphysiology platform for investigating physiology, drug safety, and disease models. Experimental Biology and Medicine 2016;241(1):101-114. R835736 (2015)
    R835736 (2016)
    R835736 (2017)
    R835736 (2018)
    R835736C004 (2015)
    R835736C004 (2016)
    R835736C004 (2017)
    R835736C004 (2018)
  • Full-text from PubMed
  • Abstract from PubMed
  • Associated PubMed link
  • Abstract: Sage Publications-Abstract
    Exit
  • Supplemental Keywords:

    SQL-SAL, sentinel cells 

    Relevant Websites:

    Microphysiology Systems Database Exit
    The Vanderbilt-Pittsburgh Resource for Organotypic Models for Predictive Toxicology Exit

    Progress and Final Reports:

    Original Abstract
  • 2016 Progress Report
  • 2017 Progress Report
  • 2018 Progress Report

  • Main Center Abstract and Reports:

    R835736    Vanderbilt Pittsburgh Resource for Organotypic Models for Predictive Toxicology

    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 LimbMalformationsResulting 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